CN111057798A - LAMP primer combination and kit for detecting novel coronavirus - Google Patents

Abstract

The invention discloses an LAMP primer combination and a kit for detecting novel coronaviruses, wherein the primer combination is selected from any one or more of a primer group 1 shown as SEQ ID NO. 3-6, a primer group 2 shown as SEQ ID NO. 7-10, a primer group 3 shown as SEQ ID NO. 11-14, a primer group 4 shown as SEQ ID NO. 15-18, a primer group 5 shown as SEQ ID NO. 19-22, a primer group 6 shown as SEQ ID NO. 24-27, a primer group 7 shown as SEQ ID NO. 28-31 and a primer group 8 shown as SEQ ID NO. 32-35. The primer combination is combined with LAMP technology, so that the detection result can be observed by naked eyes, complex auxiliary equipment is not needed, the kit is simple and easy to use, the semi-quantification of the detection result can be realized by a fluorescent dye method, the detection sensitivity is high, and a convenient and rapid means is provided for the diagnosis of the related diseases of the novel coronavirus and the research of therapeutic drugs.

Description

LAMP primer combination and kit for detecting novel coronavirus

Technical Field

The invention relates to the field of virus detection, in particular to an LAMP primer combination and a kit for detecting novel coronavirus.

Background

The gene sequence information of the novel coronavirus is published (GenBank, MN908947), and is shown as SEQ ID NO:1, so that a preliminary foundation is laid for the development of a diagnostic tool of the novel coronavirus.

According to the amplification characteristics of the strand displacement DNA polymerase and the specific primers (4 specific primers designed aiming at 6 regions of a target sequence), the strand displacement DNA polymerase is utilized to ensure that the primers are successfully combined with the template under the isothermal condition and carry out amplification reaction, the specificity and the sensitivity which are equivalent to those of polymerase chain reaction are realized, the continuous and rapid amplification under the isothermal condition can be realized, and meanwhile, the loop primers can be added, so that the amplification efficiency is improved.

Therefore, the LAMP technology is applied to the detection of the novel coronavirus, so that the related diseases can be accurately and efficiently diagnosed, and the research on the novel coronavirus is facilitated.

Disclosure of Invention

The invention provides an LAMP primer combination and a kit for detecting novel coronavirus, which are provided for solving the problems in the prior art.

In order to achieve the purpose, the invention adopts the following technical scheme:

the first aspect of the invention provides an LAMP primer combination for detecting novel coronaviruses, which is selected from any one or more of a primer group 1 shown as SEQ ID NO. 3-6, aprimer group 2 shown as SEQ ID NO. 7-10, a primer group 3 shown as SEQ ID NO. 11-14, a primer group 4 shown as SEQ ID NO. 15-18, a primer group 5 shown as SEQ ID NO. 19-22, a primer group 6 shown as SEQ ID NO. 24-27, a primer group 7 shown as SEQ ID NO. 28-31 and a primer group 8 shown as SEQ ID NO. 32-35.

The second aspect of the invention is an LAMP kit for detecting novel coronavirus, which comprises the LAMP primer combination.

Further, the concentration of the primer is 80-110. mu.M.

Further, the concentration of the primer was 100. mu.M.

Further, the LAMP kit also comprises a reaction buffer solution, BSst DNA polymerase, SYBR/neutral red dye, a DNA template and double distilled water.

Further, the reaction buffer comprises Tris-HCl, KCl, (NH)₄)₂SO₄、MgSO₄And Tween-20.

The third aspect of the present invention provides a method for detecting the novel coronavirus using the above-mentioned kit, comprising the steps of:

step 1, extracting and purifying DNA of a sample to be detected;

step 2, preparing an LAMP reaction system, and carrying out loop-mediated isothermal amplification reaction;

and step 3, judging results.

Further, the volume of the reaction system instep 2 was 30. mu.L, and the following reagents were included: 20. mu.L of reaction buffer, 0.05. mu.L of each primer, 4. mu.L of BSst DNA polymerase 1. mu. L, SYBR/neutral red dye 1. mu. L, DNA template, and the balance double distilled water.

Further, the conditions of the loop-mediated isothermal amplification reaction are 55-70 ℃ and 30-60 min.

Further preferably, the conditions of the loop-mediated isothermal amplification reaction are 63-68 ℃ and 60 min.

Even more preferably, the conditions for the loop-mediated isothermal amplification reaction are 65 ℃ for 60 min.

Furthermore, the equipment adopted by the loop-mediated isothermal amplification reaction is a common PCR instrument, a real-time PCR instrument or a constant-temperature metal bath.

By adopting the technical scheme, compared with the prior art, the invention has the following technical effects:

the LAMP primer combination for detecting the novel coronavirus is designed aiming at two target fragments of the novel coronavirus 2019-nCoV, and has strong specificity. The LAMP kit for detecting the novel coronavirus combines the primer combination with the LAMP technology, can observe a detection result by naked eyes, does not need complex auxiliary equipment, is simple and easy to use, can realize semi-quantification of the detection result by a fluorescent dye method, has high detection sensitivity, and provides a convenient and rapid means for diagnosis of related diseases of the novel coronavirus and research of therapeutic drugs.

Drawings

FIG. 1 shows the results of the visual dye method for detecting the novel coronavirus 2019-nCoV in one embodiment of the present invention;

FIG. 2 shows an amplification curve of the fluorescent dye method for detecting the novel coronavirus 2019-nCoV (A, B, C, D, E samples from left to right).

Detailed Description

The invention provides an LAMP primer combination and a kit for detecting novel coronavirus.

The present invention will be described in detail and specifically with reference to the following examples to facilitate better understanding of the present invention, but the following examples do not limit the scope of the present invention.

Example 1

An LAMP primer combination for detecting novel coronaviruses is selected from any one or more of a primer group 1 shown as SEQ ID NO. 3-6, aprimer group 2 shown as SEQ ID NO. 7-10, a primer group 3 shown as SEQ ID NO. 11-14, a primer group 4 shown as SEQ ID NO. 15-18, a primer group 5 shown as SEQ ID NO. 19-22, a primer group 6 shown as SEQ ID NO. 24-27, a primer group 7 shown as SEQ ID NO. 28-31 and a primer group 8 shown as SEQ ID NO. 32-35.

The primer sets 1-5 are designed aiming at the target sequence segment [22713-22361] of the novel coronavirus 2019-nCoV, and the sequence information of the segment is shown as SEQ ID NO. 2.

The primer group 6-8 is designed aiming at the target sequence segment [30001-30473] of the novel coronavirus 2019-nCoV, and the sequence information of the segment is shown as SEQ ID NO. 23.

Example 2

A LAMP kit for detecting a novel coronavirus comprises the LAMP primer combination as described in example 1, wherein the concentration of the primers is 100 mu M.

The LAMP kit for detecting the novel coronavirus also comprises a reaction buffer solution, BSst DNA polymerase, SYBR/neutral red dye, a DNA template and double distilled water; wherein the reaction buffer solution comprises Tris-HCl, KCl and (NH)₄)₂SO₄、MgSO₄And Tween-20.

Example 3

The method for diagnosing the novel coronavirus using the LAMP kit of example 2, comprising the steps of:

step 1, extracting and purifying DNA of a sample to be detected;

step 2, preparing an LAMP reaction system (shown in table 1), and carrying out loop-mediated isothermal amplification reaction for 60min in a constant-temperature metal bath at 65 ℃;

and step 3, judging a result: according to color or amplification curve.

Preparation of LAMP reaction System of Table 130. mu.L

Verification examples

The verification example verifies the effect of detecting the novel coronavirus by using the primer group in example 1 or the kit in example 2, and comprises the following contents:

1. case of amplification of sample

1) A novel coronavirus plasmid simulation sample with gradient concentration is disclosed, wherein the specific concentration is A: 10⁷Simulation samples of copies/ml, B: 10⁶Simulation samples of copies/ml, C: 10⁵Simulation samples of copies/ml,D: 10⁴Simulation samples of copies/ml, E: 10³copies/ml of mock sample.

2) Other types of samples: f: human genomic DNA sample, G: SARS plasmid mock sample, H: negative control samples.

2. The novel coronavirus was tested according to the procedure of example 3, and the results are shown in FIGS. 1 and 2; wherein, when a real-time fluorescence curve method is adopted, SYBR is selected as a reaction system; when the visual method is adopted, the neutral red dye is selected as the reaction system.

As is clear from FIGS. 1 and 2, the results of the visual dye method and the fluorescence curve method were consistent, A, B, C, D, E was positive, and the detection sensitivity was 10³copies/ml。

In conclusion, the LAMP primer combination and the kit for detecting the novel coronavirus provided by the invention have the advantages of sensitive detection and strong specificity.

The embodiments of the present invention have been described in detail, but the embodiments are merely examples, and the present invention is not limited to the embodiments described above. Any equivalent modifications and substitutions to those skilled in the art are also within the scope of the present invention. Accordingly, equivalent changes and modifications made without departing from the spirit and scope of the present invention should be covered by the present invention.

Sequence listing

<110> Huashan Hospital affiliated to Fudan university

<120> LAMP kit for rapid detection of novel coronavirus

<160>35

<170>SIPOSequenceListing 1.0

<210>1

<211>30473

<212>DNA

<213>2019-nCoV Gene sequence

<400>1

cggtgacgca tacaaaacat tcccaccata ccttcccagg taacaaacca accaactttc 60

gatctcttgt agatctgttc tctaaacgaa ctttaaaatc tgtgtggctg tcactcggct 120

gcatgcttag tgcactcacg cagtataatt aataactaat tactgtcgtt gacaggacac 180

gagtaactcg tctatcttct gcaggctgct tacggtttcg tccgtgttgc agccgatcat 240

cagcacatct aggtttcgtc cgggtgtgac cgaaaggtaa gatggagagc cttgtccctg 300

gtttcaacga gaaaacacac gtccaactca gtttgcctgt tttacaggtt cgcgacgtgc 360

tcgtacgtgg ctttggagac tccgtggagg aggtcttatc agaggcacgt caacatctta 420

aagatggcac ttgtggctta gtagaagttg aaaaaggcgt tttgcctcaa cttgaacagc 480

cctatgtgtt catcaaacgt tcggatgctc gaactgcacc tcatggtcat gttatggttg 540

agctggtagc agaactcgaa ggcattcagt acggtcgtag tggtgagaca cttggtgtcc 600

ttgtccctca tgtgggcgaa ataccagtgg cttaccgcaa ggttcttctt cgtaagaacg 660

gtaataaagg agctggtggc catagttacg gcgccgatct aaagtcattt gacttaggcg 720

acgagcttgg cactgatcct tatgaagatt ttcaagaaaa ctggaacact aaacatagca 780

gtggtgttac ccgtgaactc atgcgtgagc ttaacggagg ggcatacact cgctatgtcg 840

ataacaactt ctgtggccct gatggctacc ctcttgagtg cattaaagac cttctagcac 900

gtgctggtaa agcttcatgc actttgtccg aacaactgga ctttattgac actaagaggg 960

gtgtatactg ctgccgtgaa catgagcatg aaattgcttg gtacacggaa cgttctgaaa 1020

agagctatga attgcagaca ccttttgaaa ttaaattggc aaagaaattt gacaccttca 1080

atggggaatg tccaaatttt gtatttccct taaattccat aatcaagact attcaaccaa 1140

gggttgaaaa gaaaaagctt gatggcttta tgggtagaat tcgatctgtc tatccagttg 1200

cgtcaccaaa tgaatgcaac caaatgtgcc tttcaactct catgaagtgt gatcattgtg 1260

gtgaaacttc atggcagacg ggcgattttg ttaaagccac ttgcgaattt tgtggcactg 1320

agaatttgac taaagaaggt gccactactt gtggttactt accccaaaat gctgttgtta 1380

aaatttattg tccagcatgt cacaattcag aagtaggacc tgagcatagt cttgccgaat 1440

accataatga atctggcttg aaaaccattc ttcgtaaggg tggtcgcact attgcctttg 1500

gaggctgtgt gttctcttat gttggttgcc ataacaagtg tgcctattgg gttccacgtg 1560

ctagcgctaa cataggttgt aaccatacag gtgttgttgg agaaggttcc gaaggtctta 1620

atgacaacct tcttgaaata ctccaaaaag agaaagtcaa catcaatatt gttggtgact 1680

ttaaacttaa tgaagagatc gccattattt tggcatcttt ttctgcttcc acaagtgctt 1740

ttgtggaaac tgtgaaaggt ttggattata aagcattcaa acaaattgtt gaatcctgtg 1800

gtaattttaa agttacaaaa ggaaaagcta aaaaaggtgc ctggaatatt ggtgaacaga 1860

aatcaatact gagtcctctt tatgcatttg catcagaggc tgctcgtgtt gtacgatcaa 1920

ttttctcccg cactcttgaa actgctcaaa attctgtgcg tgttttacag aaggccgcta 1980

taacaatact agatggaatt tcacagtatt cactgagact cattgatgct atgatgttca 2040

catctgattt ggctactaac aatctagttg taatggccta cattacaggt ggtgttgttc 2100

agttgacttc gcagtggcta actaacatct ttggcactgt ttatgaaaaa ctcaaacccg 2160

tccttgattg gcttgaagag aagtttaagg aaggtgtaga gtttcttaga gacggttggg 2220

aaattgttaa atttatctca acctgtgctt gtgaaattgt cggtggacaa attgtcacct 2280

gtgcaaagga aattaaggag agtgttcaga cattctttaa gcttgtaaat aaatttttgg 2340

ctttgtgtgc tgactctatc attattggtg gagctaaact taaagccttg aatttaggtg 2400

aaacatttgt cacgcactca aagggattgt acagaaagtg tgttaaatcc agagaagaaa 2460

ctggcctact catgcctcta aaagccccaa aagaaattat cttcttagag ggagaaacac 2520

ttcccacaga agtgttaaca gaggaagttg tcttgaaaac tggtgattta caaccattag 2580

aacaacctac tagtgaagct gttgaagctc cattggttgg tacaccagtt tgtattaacg 2640

ggcttatgtt gctcgaaatc aaagacacag aaaagtactg tgcccttgca cctaatatga 2700

tggtaacaaa caataccttc acactcaaag gcggtgcacc aacaaaggtt acttttggtg 2760

atgacactgt gatagaagtg caaggttaca agagtgtgaa tatcactttt gaacttgatg 2820

aaaggattga taaagtactt aatgagaagt gctctgccta tacagttgaa ctcggtacag 2880

aagtaaatga gttcgcctgt gttgtggcag atgctgtcat aaaaactttg caaccagtat 2940

ctgaattact tacaccactg ggcattgatt tagatgagtg gagtatggct acatactact 3000

tatttgatga gtctggtgag tttaaattgg cttcacatat gtattgttct ttctaccctc 3060

cagatgagga tgaagaagaa ggtgattgtg aagaagaaga gtttgagcca tcaactcaat 3120

atgagtatgg tactgaagat gattaccaag gtaaaccttt ggaatttggt gccacttctg 3180

ctgctcttca acctgaagaa gagcaagaag aagattggtt agatgatgat agtcaacaaa 3240

ctgttggtca acaagacggc agtgaggaca atcagacaac tactattcaa acaattgttg 3300

aggttcaacc tcaattagag atggaactta caccagttgt tcagactatt gaagtgaata 3360

gttttagtgg ttatttaaaa cttactgaca atgtatacat taaaaatgca gacattgtgg 3420

aagaagctaa aaaggtaaaa ccaacagtgg ttgttaatgc agccaatgtt taccttaaac 3480

atggaggagg tgttgcagga gccttaaata aggctactaa caatgccatg caagttgaat 3540

ctgatgatta catagctact aatggaccac ttaaagtggg tggtagttgt gttttaagcg 3600

gacacaatct tgctaaacac tgtcttcatg ttgtcggccc aaatgttaac aaaggtgaag 3660

acattcaact tcttaagagt gcttatgaaa attttaatca gcacgaagtt ctacttgcac 3720

cattattatc agctggtatt tttggtgctg accctataca ttctttaaga gtttgtgtag 3780

atactgttcg cacaaatgtc tacttagctg tctttgataa aaatctctat gacaaacttg 3840

tttcaagctt tttggaaatg aagagtgaaa agcaagttga acaaaagatc gctgagattc 3900

ctaaagagga agttaagcca tttataactg aaagtaaacc ttcagttgaa cagagaaaac 3960

aagatgataa gaaaatcaaa gcttgtgttg aagaagttac aacaactctg gaagaaacta 4020

agttcctcac agaaaacttg ttactttata ttgacattaa tggcaatctt catccagatt 4080

ctgccactct tgttagtgac attgacatca ctttcttaaa gaaagatgct ccatatatag 4140

tgggtgatgt tgttcaagag ggtgttttaa ctgctgtggt tatacctact aaaaaggctg 4200

gtggcactac tgaaatgcta gcgaaagctt tgagaaaagt gccaacagac aattatataa 4260

ccacttaccc gggtcagggt ttaaatggtt acactgtaga ggaggcaaag acagtgctta 4320

aaaagtgtaa aagtgccttt tacattctac catctattat ctctaatgag aagcaagaaa 4380

ttcttggaac tgtttcttgg aatttgcgag aaatgcttgc acatgcagaa gaaacacgca 4440

aattaatgcc tgtctgtgtg gaaactaaag ccatagtttc aactatacag cgtaaatata 4500

agggtattaa aatacaagag ggtgtggttg attatggtgc tagattttac ttttacacca 4560

gtaaaacaac tgtagcgtca cttatcaaca cacttaacga tctaaatgaa actcttgtta 4620

caatgccact tggctatgta acacatggct taaatttgga agaagctgct cggtatatga 4680

gatctctcaa agtgccagct acagtttctg tttcttcacc tgatgctgtt acagcgtata 4740

atggttatct tacttcttct tctaaaacac ctgaagaaca ttttattgaa accatctcac 4800

ttgctggttc ctataaagat tggtcctatt ctggacaatc tacacaacta ggtatagaat 4860

ttcttaagag aggtgataaa agtgtatatt acactagtaa tcctaccaca ttccacctag 4920

atggtgaagt tatcaccttt gacaatctta agacacttct ttctttgaga gaagtgagga 4980

ctattaaggt gtttacaaca gtagacaaca ttaacctcca cacgcaagtt gtggacatgt 5040

caatgacata tggacaacag tttggtccaa cttatttgga tggagctgat gttactaaaa 5100

taaaacctca taattcacat gaaggtaaaa cattttatgt tttacctaat gatgacactc 5160

tacgtgttga ggcttttgag tactaccaca caactgatcc tagttttctg ggtaggtaca 5220

tgtcagcatt aaatcacact aaaaagtgga aatacccaca agttaatggt ttaacttcta 5280

ttaaatgggc agataacaac tgttatcttg ccactgcatt gttaacactc caacaaatag 5340

agttgaagtt taatccacct gctctacaag atgcttatta cagagcaagg gctggtgaag 5400

ctgctaactt ttgtgcactt atcttagcct actgtaataa gacagtaggt gagttaggtg 5460

atgttagaga aacaatgagt tacttgtttc aacatgccaa tttagattct tgcaaaagag 5520

tcttgaacgt ggtgtgtaaa acttgtggac aacagcagac aacccttaag ggtgtagaag 5580

ctgttatgta catgggcaca ctttcttatg aacaatttaa gaaaggtgtt cagatacctt 5640

gtacgtgtgg taaacaagct acaaaatatc tagtacaaca ggagtcacct tttgttatga 5700

tgtcagcacc acctgctcag tatgaactta agcatggtac atttacttgt gctagtgagt 5760

acactggtaa ttaccagtgt ggtcactata aacatataac ttctaaagaa actttgtatt 5820

gcatagacgg tgctttactt acaaagtcct cagaatacaa aggtcctatt acggatgttt 5880

tctacaaaga aaacagttac acaacaacca taaaaccagt tacttataaa ttggatggtg 5940

ttgtttgtac agaaattgac cctaagttgg acaattatta taagaaagac aattcttatt 6000

tcacagagca accaattgat cttgtaccaa accaaccata tccaaacgca agcttcgata 6060

attttaagtt tgtatgtgat aatatcaaat ttgctgatga tttaaaccag ttaactggtt 6120

ataagaaacc tgcttcaaga gagcttaaag ttacattttt ccctgactta aatggtgatg 6180

tggtggctat tgattataaa cactacacac cctcttttaa gaaaggagct aaattgttac 6240

ataaacctat tgtttggcat gttaacaatg caactaataa agccacgtat aaaccaaata 6300

cctggtgtat acgttgtctt tggagcacaa aaccagttga aacatcaaat tcgtttgatg 6360

tactgaagtc agaggacgcg cagggaatgg ataatcttgc ctgcgaagat ctaaaaccag 6420

tctctgaaga agtagtggaa aatcctacca tacagaaaga cgttcttgag tgtaatgtga 6480

aaactaccga agttgtagga gacattatac ttaaaccagc aaataatagt ttaaaaatta 6540

cagaagaggt tggccacaca gatctaatgg ctgcttatgt agacaattct agtcttacta 6600

ttaagaaacc taatgaatta tctagagtat taggtttgaa aacccttgct actcatggtt 6660

tagctgctgt taatagtgtc ccttgggata ctatagctaa ttatgctaag ccttttctta 6720

acaaagttgt tagtacaact actaacatag ttacacggtg tttaaaccgt gtttgtacta 6780

attatatgcc ttatttcttt actttattgc tacaattgtg tacttttact agaagtacaa 6840

attctagaat taaagcatct atgccgacta ctatagcaaa gaatactgtt aagagtgtcg 6900

gtaaattttg tctagaggct tcatttaatt atttgaagtc acctaatttt tctaaactga 6960

taaatattat aatttggttt ttactattaa gtgtttgcct aggttcttta atctactcaa 7020

ccgctgcttt aggtgtttta atgtctaatt taggcatgcc ttcttactgt actggttaca 7080

gagaaggcta tttgaactct actaatgtca ctattgcaac ctactgtact ggttctatac 7140

cttgtagtgt ttgtcttagt ggtttagatt ctttagacac ctatccttct ttagaaacta 7200

tacaaattac catttcatct tttaaatggg atttaactgc ttttggctta gttgcagagt 7260

ggtttttggc atatattctt ttcactaggt ttttctatgt acttggattg gctgcaatca 7320

tgcaattgtt tttcagctat tttgcagtac attttattag taattcttgg cttatgtggt 7380

taataattaa tcttgtacaa atggccccga tttcagctat ggttagaatg tacatcttct 7440

ttgcatcatt ttattatgta tggaaaagtt atgtgcatgt tgtagacggt tgtaattcat 7500

caacttgtat gatgtgttac aaacgtaata gagcaacaag agtcgaatgt acaactattg 7560

ttaatggtgt tagaaggtcc ttttatgtct atgctaatgg aggtaaaggc ttttgcaaac 7620

tacacaattg gaattgtgtt aattgtgata cattctgtgc tggtagtaca tttattagtg 7680

atgaagttgc gagagacttg tcactacagt ttaaaagacc aataaatcct actgaccagt 7740

cttcttacat cgttgatagt gttacagtga agaatggttc catccatctt tactttgata 7800

aagctggtca aaagacttat gaaagacatt ctctctctca ttttgttaac ttagacaacc 7860

tgagagctaa taacactaaa ggttcattgc ctattaatgt tatagttttt gatggtaaat 7920

caaaatgtga agaatcatct gcaaaatcag cgtctgttta ctacagtcag cttatgtgtc 7980

aacctatact gttactagat caggcattag tgtctgatgt tggtgatagt gcggaagttg 8040

cagttaaaat gtttgatgct tacgttaata cgttttcatc aacttttaac gtaccaatgg 8100

aaaaactcaa aacactagtt gcaactgcag aagctgaact tgcaaagaat gtgtccttag 8160

acaatgtctt atctactttt atttcagcag ctcggcaagg gtttgttgat tcagatgtag 8220

aaactaaaga tgttgttgaa tgtcttaaat tgtcacatca atctgacata gaagttactg 8280

gcgatagttg taataactat atgctcacct ataacaaagt tgaaaacatg acaccccgtg 8340

accttggtgc ttgtattgac tgtagtgcgc gtcatattaa tgcgcaggta gcaaaaagtc 8400

acaacattgc tttgatatgg aacgttaaag atttcatgtc attgtctgaa caactacgaa 8460

aacaaatacg tagtgctgct aaaaagaata acttaccttt taagttgaca tgtgcaacta 8520

ctagacaagt tgttaatgtt gtaacaacaa agatagcact taagggtggt aaaattgtta 8580

ataattggtt gaagcagtta attaaagtta cacttgtgtt cctttttgtt gctgctattt 8640

tctatttaat aacacctgtt catgtcatgt ctaaacatac tgacttttca agtgaaatca 8700

taggatacaa ggctattgat ggtggtgtca ctcgtgacat agcatctaca gatacttgtt 8760

ttgctaacaa acatgctgat tttgacacat ggtttagcca gcgtggtggt agttatacta 8820

atgacaaagc ttgcccattg attgctgcag tcataacaag agaagtgggt tttgtcgtgc 8880

ctggtttgcc tggcacgata ttacgcacaa ctaatggtga ctttttgcat ttcttaccta 8940

gagtttttag tgcagttggt aacatctgtt acacaccatc aaaacttata gagtacactg 9000

actttgcaac atcagcttgt gttttggctg ctgaatgtac aatttttaaa gatgcttctg 9060

gtaagccagt accatattgt tatgatacca atgtactaga aggttctgtt gcttatgaaa 9120

gtttacgccc tgacacacgt tatgtgctca tggatggctc tattattcaa tttcctaaca 9180

cctaccttga aggttctgtt agagtggtaa caacttttga ttctgagtac tgtaggcacg 9240

gcacttgtga aagatcagaa gctggtgttt gtgtatctac tagtggtaga tgggtactta 9300

acaatgatta ttacagatct ttaccaggag ttttctgtgg tgtagatgct gtaaatttac 9360

ttactaatat gtttacacca ctaattcaac ctattggtgc tttggacata tcagcatcta 9420

tagtagctgg tggtattgta gctatcgtag taacatgcct tgcctactat tttatgaggt 9480

ttagaagagc ttttggtgaa tacagtcatg tagttgcctt taatacttta ctattcctta 9540

tgtcattcac tgtactctgt ttaacaccag tttactcatt cttacctggt gtttattctg 9600

ttatttactt gtacttgaca ttttatctta ctaatgatgt ttctttttta gcacatattc 9660

agtggatggt tatgttcaca cctttagtac ctttctggat aacaattgct tatatcattt 9720

gtatttccac aaagcatttc tattggttct ttagtaatta cctaaagaga cgtgtagtct 9780

ttaatggtgt ttcctttagt acttttgaag aagctgcgct gtgcaccttt ttgttaaata 9840

aagaaatgta tctaaagttg cgtagtgatg tgctattacc tcttacgcaa tataatagat 9900

acttagctct ttataataag tacaagtatt ttagtggagc aatggataca actagctaca 9960

gagaagctgc ttgttgtcat ctcgcaaagg ctctcaatga cttcagtaac tcaggttctg 10020

atgttcttta ccaaccacca caaacctcta tcacctcagc tgttttgcag agtggtttta 10080

gaaaaatggc attcccatct ggtaaagttg agggttgtat ggtacaagta acttgtggta 10140

caactacact taacggtctt tggcttgatg acgtagttta ctgtccaaga catgtgatct 10200

gcacctctga agacatgctt aaccctaatt atgaagattt actcattcgt aagtctaatc 10260

ataatttctt ggtacaggct ggtaatgttc aactcagggt tattggacat tctatgcaaa 10320

attgtgtact taagcttaag gttgatacag ccaatcctaa gacacctaag tataagtttg 10380

ttcgcattca accaggacag actttttcag tgttagcttg ttacaatggt tcaccatctg 10440

gtgtttacca atgtgctatg aggcccaatt tcactattaa gggttcattc cttaatggtt 10500

catgtggtag tgttggtttt aacatagatt atgactgtgt ctctttttgt tacatgcacc 10560

atatggaatt accaactgga gttcatgctg gcacagactt agaaggtaac ttttatggac 10620

cttttgttga caggcaaaca gcacaagcag ctggtacgga cacaactatt acagttaatg 10680

ttttagcttg gttgtacgct gctgttataa atggagacag gtggtttctc aatcgattta 10740

ccacaactct taatgacttt aaccttgtgg ctatgaagta caattatgaa cctctaacac 10800

aagaccatgt tgacatacta ggacctcttt ctgctcaaac tggaattgcc gttttagata 10860

tgtgtgcttc attaaaagaa ttactgcaaa atggtatgaa tggacgtacc atattgggta 10920

gtgctttatt agaagatgaa tttacacctt ttgatgttgt tagacaatgc tcaggtgtta 10980

ctttccaaag tgcagtgaaa agaacaatca agggtacaca ccactggttg ttactcacaa 11040

ttttgacttc acttttagtt ttagtccaga gtactcaatg gtctttgttc ttttttttgt 11100

atgaaaatgc ctttttacct tttgctatgg gtattattgc tatgtctgct tttgcaatga 11160

tgtttgtcaa acataagcat gcatttctct gtttgttttt gttaccttct cttgccactg 11220

tagcttattt taatatggtc tatatgcctg ctagttgggt gatgcgtatt atgacatggt 11280

tggatatggt tgatactagt ttgtctggtt ttaagctaaa agactgtgtt atgtatgcat 11340

cagctgtagt gttactaatc cttatgacag caagaactgt gtatgatgat ggtgctagga 11400

gagtgtggac acttatgaat gtcttgacac tcgtttataa agtttattat ggtaatgctt 11460

tagatcaagc catttccatg tgggctctta taatctctgt tacttctaac tactcaggtg 11520

tagttacaac tgtcatgttt ttggccagag gtattgtttt tatgtgtgtt gagtattgcc 11580

ctattttctt cataactggt aatacacttc agtgtataat gctagtttat tgtttcttag 11640

gctatttttg tacttgttac tttggcctct tttgtttact caaccgctac tttagactga 11700

ctcttggtgt ttatgattac ttagtttcta cacaggagtt tagatatatg aattcacagg 11760

gactactccc acccaagaat agcatagatg ccttcaaact caacattaaa ttgttgggtg 11820

ttggtggcaa accttgtatc aaagtagcca ctgtacagtc taaaatgtca gatgtaaagt 11880

gcacatcagt agtcttactc tcagttttgc aacaactcag agtagaatca tcatctaaat 11940

tgtgggctca atgtgtccag ttacacaatg acattctctt agctaaagat actactgaag 12000

cctttgaaaa aatggtttca ctactttctg ttttgctttc catgcagggt gctgtagaca 12060

taaacaagct ttgtgaagaa atgctggaca acagggcaac cttacaagct atagcctcag 12120

agtttagttc ccttccatca tatgcagctt ttgctactgc tcaagaagct tatgagcagg 12180

ctgttgctaa tggtgattct gaagttgttc ttaaaaagtt gaagaagtct ttgaatgtgg 12240

ctaaatctga atttgaccgt gatgcagcca tgcaacgtaa gttggaaaag atggctgatc 12300

aagctatgac ccaaatgtat aaacaggcta gatctgagga caagagggca aaagttacta 12360

gtgctatgca gacaatgctt ttcactatgc ttagaaagtt ggataatgat gcactcaaca 12420

acattatcaa caatgcaaga gatggttgtg ttcccttgaa cataatacct cttacaacag 12480

cagccaaact aatggttgtc ataccagact ataacacata taaaaatacg tgtgatggta 12540

caacatttac ttatgcatca gcattgtggg aaatccaaca ggttgtagat gcagatagta 12600

aaattgttca acttagtgaa attagtatgg acaattcacc taatttagca tggcctctta 12660

ttgtaacagc tttaagggcc aattctgctg tcaaattaca gaataatgag cttagtcctg 12720

ttgcactacg acagatgtct tgtgctgccg gtactacaca aactgcttgc actgatgaca 12780

atgcgttagc ttactacaac acaacaaagg gaggtaggtt tgtacttgca ctgttatccg 12840

atttacagga tttgaaatgg gctagattcc ctaagagtga tggaactggt actatctata 12900

cagaactgga accaccttgt aggtttgtta cagacacacc taaaggtcct aaagtgaagt 12960

atttatactt tattaaagga ttaaacaacctaaatagagg tatggtactt ggtagtttag 13020

ctgccacagt acgtctacaa gctggtaatg caacagaagt gcctgccaat tcaactgtat 13080

tatctttctg tgcttttgct gtagatgctg ctaaagctta caaagattat ctagctagtg 13140

ggggacaacc aatcactaat tgtgttaaga tgttgtgtac acacactggt actggtcagg 13200

caataacagt tacaccggaa gccaatatgg atcaagaatc ctttggtggt gcatcgtgtt 13260

gtctgtactg ccgttgccac atagatcatc caaatcctaa aggattttgt gacttaaaag 13320

gtaagtatgt acaaatacct acaacttgtg ctaatgaccc tgtgggtttt acacttaaaa 13380

acacagtctg taccgtctgc ggtatgtgga aaggttatgg ctgtagttgt gatcaactcc 13440

gcgaacccat gcttcagtca gctgatgcac aatcgttttt aaacgggttt gcggtgtaag 13500

tgcagcccgt cttacaccgt gcggcacagg cactagtact gatgtcgtat acagggcttt 13560

tgacatctac aatgataaag tagctggttt tgctaaattc ctaaaaacta attgttgtcg 13620

cttccaagaa aaggacgaag atgacaattt aattgattct tactttgtag ttaagagaca 13680

cactttctct aactaccaac atgaagaaac aatttataat ttacttaagg attgtccagc 13740

tgttgctaaa catgacttct ttaagtttag aatagacggt gacatggtac cacatatatc 13800

acgtcaacgt cttactaaat acacaatggc agacctcgtc tatgctttaa ggcattttga 13860

tgaaggtaat tgtgacacat taaaagaaat acttgtcaca tacaattgtt gtgatgatga 13920

ttatttcaat aaaaaggact ggtatgattt tgtagaaaac ccagatatat tacgcgtata 13980

cgccaactta ggtgaacgtg tacgccaagc tttgttaaaa acagtacaat tctgtgatgc 14040

catgcgaaat gctggtattg ttggtgtact gacattagat aatcaagatc tcaatggtaa 14100

ctggtatgat ttcggtgatt tcatacaaac cacgccaggt agtggagttc ctgttgtaga 14160

ttcttattat tcattgttaa tgcctatatt aaccttgacc agggctttaa ctgcagagtc 14220

acatgttgac actgacttaa caaagcctta cattaagtgg gatttgttaa aatatgactt 14280

cacggaagag aggttaaaac tctttgaccg ttattttaaa tattgggatc agacatacca 14340

cccaaattgt gttaactgtt tggatgacag atgcattctg cattgtgcaa actttaatgt 14400

tttattctct acagtgttcc cacctacaag ttttggacca ctagtgagaa aaatatttgt 14460

tgatggtgtt ccatttgtag tttcaactgg ataccacttc agagagctag gtgttgtaca 14520

taatcaggat gtaaacttac atagctctag acttagtttt aaggaattac ttgtgtatgc 14580

tgctgaccct gctatgcacg ctgcttctgg taatctatta ctagataaac gcactacgtg 14640

cttttcagta gctgcactta ctaacaatgt tgcttttcaa actgtcaaac ccggtaattt 14700

taacaaagac ttctatgact ttgctgtgtc taagggtttc tttaaggaag gaagttctgt 14760

tgaattaaaa cacttcttct ttgctcagga tggtaatgct gctatcagcg attatgacta 14820

ctatcgttat aatctaccaa caatgtgtga tatcagacaa ctactatttg tagttgaagt 14880

tgttgataag tactttgatt gttacgatgg tggctgtatt aatgctaacc aagtcatcgt 14940

caacaaccta gacaaatcag ctggttttcc atttaataaa tggggtaagg ctagacttta 15000

ttatgattca atgagttatg aggatcaaga tgcacttttc gcatatacaa aacgtaatgt 15060

catccctact ataactcaaa tgaatcttaa gtatgccatt agtgcaaaga atagagctcg 15120

caccgtagct ggtgtctcta tctgtagtac tatgaccaat agacagtttc atcaaaaatt 15180

attgaaatca atagccgcca ctagaggagc tactgtagta attggaacaa gcaaattcta 15240

tggtggttgg cacaacatgt taaaaactgt ttatagtgat gtagaaaacc ctcaccttat 15300

gggttgggat tatcctaaat gtgatagagc catgcctaac atgcttagaa ttatggcctc 15360

acttgttctt gctcgcaaac atacaacgtg ttgtagcttg tcacaccgtt tctatagatt 15420

agctaatgag tgtgctcaag tattgagtga aatggtcatg tgtggcggtt cactatatgt 15480

taaaccaggt ggaacctcat caggagatgc cacaactgct tatgctaata gtgtttttaa 15540

catttgtcaa gctgtcacgg ccaatgttaa tgcactttta tctactgatg gtaacaaaat 15600

tgccgataag tatgtccgca atttacaaca cagactttat gagtgtctct atagaaatag 15660

agatgttgac acagactttg tgaatgagtt ttacgcatat ttgcgtaaac atttctcaat 15720

gatgatactc tctgacgatg ctgttgtgtg tttcaatagc acttatgcat ctcaaggtct 15780

agtggctagc ataaagaact ttaagtcagt tctttattat caaaacaatg tttttatgtc 15840

tgaagcaaaa tgttggactg agactgacct tactaaagga cctcatgaat tttgctctca 15900

acatacaatg ctagttaaac agggtgatga ttatgtgtac cttccttacc cagatccatc 15960

aagaatccta ggggccggct gttttgtaga tgatatcgta aaaacagatg gtacacttat 16020

gattgaacgg ttcgtgtctt tagctataga tgcttaccca cttactaaac atcctaatca 16080

ggagtatgct gatgtctttc atttgtactt acaatacata agaaagctac atgatgagtt 16140

aacaggacac atgttagaca tgtattctgt tatgcttact aatgataaca cttcaaggta 16200

ttgggaacct gagttttatgaggctatgta cacaccgcat acagtcttac aggctgttgg 16260

ggcttgtgtt ctttgcaatt cacagacttc attaagatgt ggtgcttgca tacgtagacc 16320

attcttatgt tgtaaatgct gttacgacca tgtcatatca acatcacata aattagtctt 16380

gtctgttaat ccgtatgttt gcaatgctcc aggttgtgat gtcacagatg tgactcaact 16440

ttacttagga ggtatgagct attattgtaa atcacataaa ccacccatta gttttccatt 16500

gtgtgctaat ggacaagttt ttggtttata taaaaataca tgtgttggta gcgataatgt 16560

tactgacttt aatgcaattg caacatgtga ctggacaaat gctggtgatt acattttagc 16620

taacacctgt actgaaagac tcaagctttt tgcagcagaa acgctcaaag ctactgagga 16680

gacatttaaa ctgtcttatg gtattgctac tgtacgtgaa gtgctgtctg acagagaatt 16740

acatctttca tgggaagttg gtaaacctag accaccactt aaccgaaatt atgtctttac 16800

tggttatcgt gtaactaaaa acagtaaagt acaaatagga gagtacacct ttgaaaaagg 16860

tgactatggt gatgctgttg tttaccgagg tacaacaact tacaaattaa atgttggtga 16920

ttattttgtg ctgacatcac atacagtaat gccattaagt gcacctacac tagtgccaca 16980

agagcactat gttagaatta ctggcttata cccaacactc aatatctcag atgagttttc 17040

tagcaatgtt gcaaattatc aaaaggttgg tatgcaaaag tattctacac tccagggacc 17100

acctggtact ggtaagagtc attttgctat tggcctagct ctctactacc cttctgctcg 17160

catagtgtat acagcttgct ctcatgccgc tgttgatgca ctatgtgaga aggcattaaa 17220

atatttgcct atagataaat gtagtagaat tatacctgca cgtgctcgtg tagagtgttt 17280

tgataaattc aaagtgaatt caacattaga acagtatgtc ttttgtactg taaatgcatt 17340

gcctgagacg acagcagata tagttgtctt tgatgaaatt tcaatggcca caaattatga 17400

tttgagtgtt gtcaatgcca gattacgtgc taagcactat gtgtacattg gcgaccctgc 17460

tcaattacct gcaccacgca cattgctaac taagggcaca ctagaaccag aatatttcaa 17520

ttcagtgtgt agacttatga aaactatagg tccagacatg ttcctcggaa cttgtcggcg 17580

ttgtcctgct gaaattgttg acactgtgag tgctttggtt tatgataata agcttaaagc 17640

acataaagac aaatcagctc aatgctttaa aatgttttat aagggtgtta tcacgcatga 17700

tgtttcatct gcaattaaca ggccacaaat aggcgtggta agagaattcc ttacacgtaa 17760

ccctgcttgg agaaaagctg tctttatttc accttataat tcacagaatg ctgtagcctc 17820

aaagattttg ggactaccaa ctcaaactgt tgattcatca cagggctcag aatatgacta 17880

tgtcatattc actcaaacca ctgaaacagc tcactcttgt aatgtaaaca gatttaatgt 17940

tgctattacc agagcaaaag taggcatact ttgcataatg tctgatagag acctttatga 18000

caagttgcaa tttacaagtc ttgaaattcc acgtaggaat gtggcaactt tacaagctga 18060

aaatgtaaca ggactcttta aagattgtag taaggtaatc actgggttac atcctacaca 18120

ggcacctaca cacctcagtg ttgacactaa attcaaaact gaaggtttat gtgttgacat 18180

acctggcata cctaaggaca tgacctatag aagactcatc tctatgatgg gttttaaaat 18240

gaattatcaa gttaatggtt accctaacat gtttatcacc cgcgaagaag ctataagaca 18300

tgtacgtgca tggattggct tcgatgtcga ggggtgtcat gctactagag aagctgttgg 18360

taccaattta cctttacagc taggtttttc tacaggtgtt aacctagttg ctgtacctac 18420

aggttatgtt gatacaccta ataatacaga tttttccaga gttagtgcta aaccaccgcc 18480

tggagatcaa tttaaacacc tcataccact tatgtacaaa ggacttcctt ggaatgtagt 18540

gcgtataaag attgtacaaa tgttaagtga cacacttaaa aatctctctg acagagtcgt 18600

atttgtctta tgggcacatg gctttgagtt gacatctatg aagtattttg tgaaaatagg 18660

acctgagcgc acctgttgtc tatgtgatag acgtgccaca tgcttttcca ctgcttcaga 18720

cacttatgcc tgttggcatc attctattgg atttgattac gtctataatc cgtttatgat 18780

tgatgttcaa caatggggtt ttacaggtaa cctacaaagc aaccatgatc tgtattgtca 18840

agtccatggt aatgcacatg tagctagttg tgatgcaatc atgactaggt gtctagctgt 18900

ccacgagtgc tttgttaagc gtgttgactg gactattgaa tatcctataa ttggtgatga 18960

actgaagatt aatgcggctt gtagaaaggt tcaacacatg gttgttaaag ctgcattatt 19020

agcagacaaa ttcccagttc ttcacgacat tggtaaccct aaagctatta agtgtgtacc 19080

tcaagctgat gtagaatgga agttctatga tgcacagcct tgtagtgaca aagcttataa 19140

aatagaagaa ttattctatt cttatgccac acattctgac aaattcacag atggtgtatg 19200

cctattttgg aattgcaatg tcgatagata tcctgctaat tccattgttt gtagatttga 19260

cactagagtg ctatctaacc ttaacttgcc tggttgtgat ggtggcagtt tgtatgtaaa 19320

taaacatgca ttccacacac cagcttttga taaaagtgct tttgttaatt taaaacaatt 19380

accatttttc tattactctg acagtccatg tgagtctcat ggaaaacaag tagtgtcaga 19440

tatagattat gtaccactaa agtctgctac gtgtataaca cgttgcaatt taggtggtgc 19500

tgtctgtaga catcatgcta atgagtacag attgtatctc gatgcttata acatgatgat 19560

ctcagctggc tttagcttgt gggtttacaa acaatttgat acttataacc tctggaacac 19620

ttttacaaga cttcagagtt tagaaaatgt ggcttttaat gttgtaaata agggacactt 19680

tgatggacaa cagggtgaag taccagtttc tatcattaat aacactgttt acacaaaagt 19740

tgatggtgtt gatgtagaat tgtttgaaaa taaaacaaca ttacctgtta atgtagcatt 19800

tgagctttgg gctaagcgca acattaaacc agtaccagag gtgaaaatac tcaataattt 19860

gggtgtggac attgctgcta atactgtgat ctgggactac aaaagagatg ctccagcaca 19920

tatatctact attggtgttt gttctatgac tgacatagcc aagaaaccaa ctgaaacgat 19980

ttgtgcacca ctcactgtct tttttgatgg tagagttgat ggtcaagtag acttatttag 20040

aaatgcccgt aatggtgttc ttattacaga aggtagtgtt aaaggtttac aaccatctgt 20100

aggtcccaaa caagctagtc ttaatggagt cacattaatt ggagaagccg taaaaacaca 20160

gttcaattat tataagaaag ttgatggtgt tgtccaacaa ttacctgaaa cttactttac 20220

tcagagtaga aatttacaag aatttaaacc caggagtcaa atggaaattg atttcttaga 20280

attagctatg gatgaattca ttgaacggta taaattagaa ggctatgcct tcgaacatat 20340

cgtttatgga gattttagtc atagtcagtt aggtggttta catctactga ttggactagc 20400

taaacgtttt aaggaatcac cttttgaatt agaagatttt attcctatgg acagtacagt 20460

taaaaactat ttcataacag atgcgcaaac aggttcatct aagtgtgtgt gttctgttat 20520

tgatttatta cttgatgatt ttgttgaaat aataaaatcc caagatttat ctgtagtttc 20580

taaggttgtc aaagtgacta ttgactatac agaaatttca tttatgcttt ggtgtaaaga 20640

tggccatgta gaaacatttt acccaaaatt acaatctagt caagcgtggc aaccgggtgt 20700

tgctatgcct aatctttaca aaatgcaaag aatgctatta gaaaagtgtg accttcaaaa 20760

ttatggtgat agtgcaacat tacctaaagg cataatgatg aatgtcgcaa aatatactca 20820

actgtgtcaa tatttaaaca cattaacatt agctgtaccc tataatatga gagttataca 20880

ttttggtgct ggttctgata aaggagttgc accaggtaca gctgttttaa gacagtggtt 20940

gcctacgggt acgctgcttg tcgattcaga tcttaatgac tttgtctctg atgcagattc 21000

aactttgatt ggtgattgtg caactgtaca tacagctaat aaatgggatc tcattattag 21060

tgatatgtac gaccctaaga ctaaaaatgt tacaaaagaa aatgactcta aagagggttt 21120

tttcacttac atttgtgggt ttatacaaca aaagctagct cttggaggtt ccgtggctat 21180

aaagataaca gaacattctt ggaatgctga tctttataag ctcatgggac acttcgcatg 21240

gtggacagcc tttgttacta atgtgaatgc gtcatcatct gaagcatttt taattggatg 21300

taattatctt ggcaaaccac gcgaacaaat agatggttat gtcatgcatg caaattacat 21360

attttggagg aatacaaatc caattcagtt gtcttcctat tctttatttg acatgagtaa 21420

atttcccctt aaattaaggg gtactgctgt tatgtcttta aaagaaggtc aaatcaatga 21480

tatgatttta tctcttctta gtaaaggtag acttataatt agagaaaaca acagagttgt 21540

tatttctagt gatgttcttg ttaacaacta aacgaacaat gtttgttttt cttgttttat 21600

tgccactagt ctctagtcag tgtgttaatc ttacaaccag aactcaatta ccccctgcat 21660

acactaattc tttcacacgt ggtgtttatt accctgacaa agttttcaga tcctcagttt 21720

tacattcaac tcaggacttg ttcttacctt tcttttccaa tgttacttgg ttccatgcta 21780

tacatgtctc tgggaccaat ggtactaaga ggtttgataa ccctgtccta ccatttaatg 21840

atggtgttta ttttgcttcc actgagaagt ctaacataat aagaggctgg atttttggta 21900

ctactttaga ttcgaagacc cagtccctac ttattgttaa taacgctact aatgttgtta 21960

ttaaagtctg tgaatttcaa ttttgtaatg atccattttt gggtgtttat taccacaaaa 22020

acaacaaaag ttggatggaa agtgagttca gagtttattc tagtgcgaat aattgcactt 22080

ttgaatatgt ctctcagcct tttcttatgg accttgaagg aaaacagggt aatttcaaaa 22140

atcttaggga atttgtgttt aagaatattg atggttattt taaaatatat tctaagcaca 22200

cgcctattaa tttagtgcgt gatctccctc agggtttttc ggctttagaa ccattggtag 22260

atttgccaat aggtattaac atcactaggt ttcaaacttt acttgcttta catagaagtt 22320

atttgactcc tggtgattct tcttcaggtt ggacagctgg tgctgcagct tattatgtgg 22380

gttatcttca acctaggact tttctattaa aatataatga aaatggaacc attacagatg 22440

ctgtagactg tgcacttgac cctctctcag aaacaaagtg tacgttgaaa tccttcactg 22500

tagaaaaagg aatctatcaa acttctaact ttagagtcca accaacagaa tctattgtta 22560

gatttcctaa tattacaaac ttgtgccctt ttggtgaagt ttttaacgcc accagatttg 22620

catctgttta tgcttggaac aggaagagaa tcagcaactg tgttgctgat tattctgtcc 22680

tatataattc cgcatcattt tccactttta agtgttatgg agtgtctcct actaaattaa 22740

atgatctctg ctttactaat gtctatgcag attcatttgt aattagaggt gatgaagtca 22800

gacaaatcgc tccagggcaa actggaaaga ttgctgatta taattataaa ttaccagatg 22860

attttacagg ctgcgttata gcttggaatt ctaacaatct tgattctaag gttggtggta 22920

attataatta cctgtataga ttgtttagga agtctaatct caaacctttt gagagagata 22980

tttcaactga aatctatcag gccggtagca caccttgtaa tggtgttgaa ggttttaatt 23040

gttactttcc tttacaatca tatggtttcc aacccactaa tggtgttggt taccaaccat 23100

acagagtagt agtactttct tttgaacttc tacatgcacc agcaactgtt tgtggaccta 23160

aaaagtctac taatttggtt aaaaacaaat gtgtcaattt caacttcaat ggtttaacag 23220

gcacaggtgt tcttactgag tctaacaaaa agtttctgcc tttccaacaa tttggcagag 23280

acattgctga cactactgat gctgtccgtg atccacagac acttgagatt cttgacatta 23340

caccatgttc ttttggtggt gtcagtgtta taacaccagg aacaaatact tctaaccagg 23400

ttgctgttct ttatcaggat gttaactgca cagaagtccc tgttgctatt catgcagatc 23460

aacttactcc tacttggcgt gtttattcta caggttctaa tgtttttcaa acacgtgcag 23520

gctgtttaat aggggctgaa catgtcaaca actcatatga gtgtgacata cccattggtg 23580

caggtatatg cgctagttat cagactcaga ctaattctcc tcggcgggca cgtagtgtag 23640

ctagtcaatc catcattgcc tacactatgt cacttggtgc agaaaattca gttgcttact 23700

ctaataactc tattgccata cccacaaatt ttactattag tgttaccaca gaaattctac 23760

cagtgtctat gaccaagaca tcagtagatt gtacaatgta catttgtggt gattcaactg 23820

aatgcagcaa tcttttgttg caatatggca gtttttgtac acaattaaac cgtgctttaa 23880

ctggaatagc tgttgaacaa gacaaaaaca cccaagaagt ttttgcacaa gtcaaacaaa 23940

tttacaaaac accaccaatt aaagattttg gtggttttaa tttttcacaa atattaccag 24000

atccatcaaa accaagcaag aggtcattta ttgaagatct acttttcaac aaagtgacac 24060

ttgcagatgc tggcttcatc aaacaatatg gtgattgcct tggtgatatt gctgctagag 24120

acctcatttg tgcacaaaag tttaacggcc ttactgtttt gccacctttg ctcacagatg 24180

aaatgattgc tcaatacact tctgcactgt tagcgggtac aatcacttct ggttggacct 24240

ttggtgcagg tgctgcatta caaataccat ttgctatgca aatggcttat aggtttaatg 24300

gtattggagt tacacagaat gttctctatg agaaccaaaa attgattgcc aaccaattta 24360

atagtgctat tggcaaaatt caagactcac tttcttccac agcaagtgca cttggaaaac 24420

ttcaagatgt ggtcaaccaa aatgcacaag ctttaaacac gcttgttaaa caacttagct 24480

ccaattttgg tgcaatttca agtgttttaa atgatatcct ttcacgtctt gacaaagttg 24540

aggctgaagt gcaaattgat aggttgatca caggcagact tcaaagtttg cagacatatg 24600

tgactcaaca attaattaga gctgcagaaa tcagagcttc tgctaatctt gctgctacta 24660

aaatgtcaga gtgtgtactt ggacaatcaa aaagagttga tttttgtgga aagggctatc 24720

atcttatgtc cttccctcag tcagcacctc atggtgtagt cttcttgcat gtgacttatg 24780

tccctgcaca agaaaagaac ttcacaactg ctcctgccat ttgtcatgat ggaaaagcac 24840

actttcctcg tgaaggtgtc tttgtttcaa atggcacaca ctggtttgta acacaaagga 24900

atttttatga accacaaatc attactacag acaacacatt tgtgtctggt aactgtgatg 24960

ttgtaatagg aattgtcaac aacacagttt atgatccttt gcaacctgaa ttagactcat 25020

tcaaggagga gttagataaa tattttaaga atcatacatc accagatgtt gatttaggtg 25080

acatctctgg cattaatgct tcagttgtaa acattcaaaa agaaattgac cgcctcaatg 25140

aggttgccaa gaatttaaat gaatctctca tcgatctcca agaacttgga aagtatgagc 25200

agtatataaa atggccatgg tacatttggc taggttttat agctggcttg attgccatag 25260

taatggtgac aattatgctt tgctgtatga ccagttgctg tagttgtctc aagggctgtt 25320

gttcttgtgg atcctgctgc aaatttgatg aagacgactc tgagccagtg ctcaaaggag 25380

tcaaattaca ttacacataa acgaacttat ggatttgttt atgagaatct tcacaattgg 25440

aactgtaact ttgaagcaag gtgaaatcaa ggatgctact ccttcagatt ttgttcgcgc 25500

tactgcaacg ataccgatac aagcctcact ccctttcgga tggcttattg ttggcgttgc 25560

acttcttgct gtttttcaga gcgcttccaa aatcataacc ctcaaaaaga gatggcaact 25620

agcactctcc aagggtgttc actttgtttg caacttgctg ttgttgtttg taacagttta 25680

ctcacacctt ttgctcgttg ctgctggcct tgaagcccct tttctctatc tttatgcttt 25740

agtctacttc ttgcagagta taaactttgt aagaataata atgaggcttt ggctttgctg 25800

gaaatgccgt tccaaaaacc cattacttta tgatgccaac tattttcttt gctggcatac 25860

taattgttac gactattgta taccttacaa tagtgtaact tcttcaattg tcattacttc 25920

aggtgatggc acaacaagtc ctatttctga acatgactac cagattggtg gttatactga 25980

aaaatgggaa tctggagtaa aagactgtgt tgtattacac agttacttca cttcagacta 26040

ttaccagctg tactcaactc aattgagtac agacactggt gttgaacatg ttaccttctt 26100

catctacaat aaaattgttg atgagcctga agaacatgtc caaattcaca caatcgacgg 26160

ttcatccgga gttgttaatc cagtaatgga accaatttat gatgaaccga cgacgactac 26220

tagcgtgcct ttgtaagcac aagctgatga gtacgaactt atgtactcat tcgtttcgga 26280

agagacaggt acgttaatag ttaatagcgt acttcttttt cttgctttcg tggtattctt 26340

gctagttaca ctagccatcc ttactgcgct tcgattgtgt gcgtactgct gcaatattgt 26400

taacgtgagt cttgtaaaac cttcttttta cgtttactct cgtgttaaaa atctgaattc 26460

ttctagagtt cctgatcttc tggtctaaac gaactaaata ttatattagt ttttctgttt 26520

ggaactttaa ttttagccat ggcagattcc aacggtacta ttaccgttga agagcttaaa 26580

aagctccttg aacaatggaa cctagtaata ggtttcctat tccttacatg gatttgtctt 26640

ctacaatttg cctatgccaa caggaatagg tttttgtata taattaagtt aattttcctc 26700

tggctgttat ggccagtaac tttagcttgt tttgtgcttg ctgctgttta cagaataaat 26760

tggatcaccg gtggaattgc tatcgcaatg gcttgtcttg taggcttgat gtggctcagc 26820

tacttcattg cttctttcag actgtttgcg cgtacgcgtt ccatgtggtc attcaatcca 26880

gaaactaaca ttcttctcaa cgtgccactc catggcacta ttctgaccag accgcttcta 26940

gaaagtgaac tcgtaatcgg agctgtgatc cttcgtggac atcttcgtat tgctggacac 27000

catctaggac gctgtgacat caaggacctg cctaaagaaa tcactgttgc tacatcacga 27060

acgctttctt attacaaatt gggagcttcg cagcgtgtag caggtgactc aggttttgct 27120

gcatacagtc gctacaggat tggcaactat aaattaaaca cagaccattc cagtagcagt 27180

gacaatattg ctttgcttgt acagtaagtg acaacagatg tttcatctcg ttgactttca 27240

ggttactata gcagagatat tactaattat tatgaggact tttaaagttt ccatttggaa 27300

tcttgattac atcataaacc tcataattaa aaatttatct aagtcactaa ctgagaataa 27360

atattctcaa ttagatgaag agcaaccaat ggagattgat taaacgaaca tgaaaattat 27420

tcttttcttg gcactgataa cactcgctac ttgtgagctt tatcactacc aagagtgtgt 27480

tagaggtaca acagtacttt taaaagaacc ttgctcttct ggaacatacg agggcaattc 27540

accatttcat cctctagctg ataacaaatt tgcactgact tgctttagca ctcaatttgc 27600

ttttgcttgt cctgacggcg taaaacacgt ctatcagtta cgtgccagat cagtttcacc 27660

taaactgttc atcagacaag aggaagttca agaactttac tctccaattt ttcttattgt 27720

tgcggcaata gtgtttataa cactttgctt cacactcaaa agaaagacag aatgattgaa 27780

ctttcattaa ttgacttcta tttgtgcttt ttagcctttc tgctattcct tgttttaatt 27840

atgcttatta tcttttggtt ctcacttgaa ctgcaagatc ataatgaaac ttgtcacgcc 27900

taaacgaaca tgaaatttct tgttttctta ggaatcatca caactgtagc tgcatttcac 27960

caagaatgta gtttacagtc atgtactcaa catcaaccat atgtagttga tgacccgtgt 28020

cctattcact tctattctaa atggtatatt agagtaggag ctagaaaatc agcaccttta 28080

attgaattgt gcgtggatga ggctggttct aaatcaccca ttcagtacat cgatatcggt 28140

aattatacag tttcctgttt accttttaca attaattgcc aggaacctaa attgggtagt 28200

cttgtagtgc gttgttcgtt ctatgaagac tttttagagt atcatgacgt tcgtgttgtt 28260

ttagatttca tctaaacgaa caaactaaaa tgtctgataa tggaccccaa aatcagcgaa 28320

atgcaccccg cattacgttt ggtggaccct cagattcaac tggcagtaac cagaatggag 28380

aacgcagtgg ggcgcgatca aaacaacgtc ggccccaagg tttacccaat aatactgcgt 28440

cttggttcac cgctctcact caacatggca aggaagacct taaattccct cgaggacaag 28500

gcgttccaat taacaccaat agcagtccag atgaccaaat tggctactac cgaagagcta 28560

ccagacgaat tcgtggtggt gacggtaaaa tgaaagatct cagtccaaga tggtatttct 28620

actacctagg aactgggcca gaagctggac ttccctatgg tgctaacaaa gacggcatca 28680

tatgggttgc aactgaggga gccttgaata caccaaaaga tcacattggc acccgcaatc 28740

ctgctaacaa tgctgcaatc gtgctacaac ttcctcaagg aacaacattg ccaaaaggct 28800

tctacgcaga agggagcaga ggcggcagtc aagcctcttc tcgttcctca tcacgtagtc 28860

gcaacagttc aagaaattca actccaggca gcagtagggg aacttctcct gctagaatgg 28920

ctggcaatgg cggtgatgct gctcttgctt tgctgctgct tgacagattg aaccagcttg 28980

agagcaaaat gtctggtaaa ggccaacaac aacaaggcca aactgtcact aagaaatctg 29040

ctgctgaggc ttctaagaag cctcggcaaa aacgtactgc cactaaagca tacaatgtaa 29100

cacaagcttt cggcagacgt ggtccagaac aaacccaagg aaattttggg gaccaggaac 29160

taatcagaca aggaactgat tacaaacatt ggccgcaaat tgcacaattt gcccccagcg 29220

cttcagcgtt cttcggaatg tcgcgcattg gcatggaagt cacaccttcg ggaacgtggt 29280

tgacctacac agctgccatc aaattggatg acaaagatcc aaatttcaaa gatcaagtca 29340

ttttgctgaa taagcatatt gacgcataca aaacattccc accaacagag cctaaaaagg 29400

acaaaaagaa gaaggctgat gaaactcaag ccttaccgca gagacagaag aaacagcaaa 29460

ctgtgactct tcttcctgct gcagatttgg atgatttctc caaacaattg caacaatcca 29520

tgagcagtgc tgactcaact caggcctaaa ctcatgcaga ccacacaagg cagatgggct 29580

atataaacgt tttcgctttt ccgtttacga tatatagtct actcttgtgc agaatgaatt 29640

ctcgtaacta catagcacaa gtagatgtag ttaactttaa tctcacatag caatctttaa 29700

tcagtgtgta acattaggga ggacttgaaa gagccaccac attttcaccg aggccacgcg 29760

gagtacgatc gagtgtacag tgaacaatgc tagggagagc tgcctatatg gaagagccct 29820

aatgtgtaaa attaatttta gtagtgctat ccccatgtga ttttaatagc ttcttcctgg 29880

gatcttgatt tcaacagcct cttcctcata ctattctcaa cactactgtc agtgaacttc 29940

taaaaatgga ttctgtgttt gaccgtagga aacatttcag tattccttat ctttagaaac 30000

ctctgcaact caagtgtctc tgccaaagag tcttcaaggt aatgtcaaat accataacat 30060

attttttaag tttttgtata cttcctagga atatatgtca tagtatgtaa agaagatctg 30120

attcatgtga tctgtactgt taagaaacac ttcaaaggaa aacaacctct catagatctt 30180

caaatcctgt tgttcttgag ttggaatgta caatattcag gtagagatgc tggtacaaaa 30240

aaagactgaa tcatcaggct attagaaaga taaactaggc ctctaacata gtaactttaa 30300

cagtttgtac ttatacatat tttcacattg aaatatagtt ttattcatga ctttttttgt 30360

tttagcttct ctgtcttcca ttatttcaag ctgctaaaaa ttaaaaatat cctatagcaa 30420

agggctatgg catctttttg taaaaataag gaaagcaagg ttttttgata atc 30473

<210>2

<211>577

<212>DNA

<213> target sequence fragment [22713-22361]

<400>2

ctcagtttta cattcaactc aggacttgtt cttacctttc ttttccaatg ttacttggtt 60

ccatgctata catgtctctg ggaccaatgg tactaagagg tttgataacc ctgtcctacc 120

atttaatgat ggtgtttatt ttgcttccac tgagaagtct aacataataa gaggctggat 180

ttttggtact actttagatt cgaagaccca gtccctactt attgttaata acgctactaa 240

tgttgttatt aaagtctgtg aatttcaatt ttgtaatgat ccatttttgg gtgtttatta 300

ccacaaaaac aacaaaagtt ggatggaaag tgagttcaga gtttattcta gtgcgaataa 360

ttgcactttt gaatatgtct ctcagccttt tcttatggac cttgaaggaa aacagggtaa 420

tttcaaaaat cttagggaat ttgtgtttaa gaatattgat ggttatttta aaatatattc 480

taagcacacg cctattaatt tagtgcgtga tctccctcag ggtttttcgg ctttagaacc 540

attggtagat ttgccaatag gtattaacat cactagg 577

<210>3

<211>20

<212>DNA

<213> combination 1F 3 sequence

<400>3

tcaactcagg acttgttctt 20

<210>4

<211>18

<212>DNA

<213> combination 1B 3 sequence

<400>4

ggactgggtc ttcgaatc 18

<210>5

<211>46

<212>DNA

<213> Combined 1 FIP sequences

<400>5

acctcttagt accattggtc ccactttctt ttccaatgtt acttgg 46

<210>6

<211>42

<212>DNA

<213> combination 1 BIP sequences

<400>6

cctgtcctac catttaatga tggtaccaaa aatccagcct ct 42

<210>7

<211>21

<212>DNA

<213> Combined 2F 3 sequences

<400>7

caggacttgt tcttaccttt c 21

<210>8

<211>24

<212>DNA

<213> Combined 2B 3 sequences

<400>8

ttattaacaa taagtagggactgg 24

<210>9

<211>46

<212>DNA

<213> Combined 2 FIP sequences

<400>9

aggacagggt tatcaaacct cttagcaatg ttacttggtt ccatgc 46

<210>10

<211>46

<212>DNA

<213> Combined 2 BIP sequences

<400>10

atgatggtgt ttattttgct tccacaagta gtaccaaaaa tccagc 46

<210>11

<211>23

<212>DNA

<213> Combined 3F 3 sequences

<400>11

ctttcttttc caatgttact tgg 23

<210>12

<211>25

<212>DNA

<213> combinatorial 3B 3 sequences

<400>12

gactttaata acaacattag tagcg 25

<210>13

<211>47

<212>DNA

<213> Combined 3 FIP sequences

<400>13

catcattaaa tggtaggaca gggttttcca tgctatacat gtctctg 47

<210>14

<211>41

<212>DNA

<213> Combined 3 BIP sequences

<400>14

tgcttccact gagaagtcta acaggactgg gtcttcgaat c 41

<210>15

<211>21

<212>DNA

<213> combinatorial 4F 3 sequences

<400>15

ccatgctata catgtctctg g 21

<210>16

<211>20

<212>DNA

<213> combinatorial 4B 3 sequences

<400>16

aaacacccaa aaatggatca 20

<210>17

<211>45

<212>DNA

<213> Combined 4 FIP sequences

<400>17

tggaagcaaa ataaacacca tcattgacca atggtactaa gaggt 45

<210>18

<211>50

<212>DNA

<213> combinatorial 4 BIP sequences

<400>18

actactttag attcgaagac ccagtgactt taataacaac attagtagcg 50

<210>19

<211>18

<212>DNA

<213> combinatorial 5F 3 sequences

<400>19

gtctctggga ccaatggt 18

<210>20

<211>20

<212>DNA

<213> combinatorial 5B 3 sequences

<400>20

aaacacccaa aaatggatca 20

<210>21

<211>48

<212>DNA

<213> Association 5 FIP sequences

<400>21

ttagacttct cagtggaagc aaaatctaag aggtttgata accctgtc 48

<210>22

<211>50

<212>DNA

<213> Association 5 BIP sequences

<400>22

actactttag attcgaagac ccagtgactt taataacaac attagtagcg 50

<210>23

<211>473

<212>DNA

<213> target fragment [30001-30473]

<400>23

ctctgcaact caagtgtctc tgccaaagag tcttcaaggt aatgtcaaat accataacat 60

attttttaag tttttgtata cttcctagga atatatgtca tagtatgtaa agaagatctg 120

attcatgtga tctgtactgt taagaaacac ttcaaaggaa aacaacctct catagatctt 180

caaatcctgt tgttcttgag ttggaatgta caatattcag gtagagatgc tggtacaaaa 240

aaagactgaa tcatcaggct attagaaaga taaactaggc ctctaacata gtaactttaa 300

cagtttgtac ttatacatat tttcacattg aaatatagtt ttattcatga ctttttttgt 360

tttagcttct ctgtcttcca ttatttcaag ctgctaaaaa ttaaaaatat cctatagcaa 420

agggctatgg catctttttg taaaaataag gaaagcaagg ttttttgata atc 473

<210>24

<211>25

<212>DNA

<213> combinatorial 6F 3 sequences

<400>24

gtcatagtat gtaaagaaga tctga 25

<210>25

<211>25

<212>DNA

<213> combinatorial 6B 3 sequences

<400>25

actgttaaag ttactatgtt agagg 25

<210>26

<211>49

<212>DNA

<213> Association 6 FIP sequences

<400>26

ccaactcaag aacaacagga tttgatgtga tctgtactgt taagaaaca 49

<210>27

<211>50

<212>DNA

<213> combinatorial 6 BIP sequences

<400>27

acaatattca ggtagagatg ctggtagttt atctttctaa tagcctgatg 50

<210>28

<211>22

<212>DNA

<213> combinatorial 7F 3 sequences

<400>28

agatctgatt catgtgatct gt 22

<210>29

<211>25

<212>DNA

<213> combinatorial 7B 3 sequences

<400>29

actgttaaag ttactatgtt agagg 25

<210>30

<211>48

<212>DNA

<213> Association 7 FIP sequences

<400>30

ccaactcaag aacaacagga tttgactgtt aagaaacact tcaaagga 48

<210>31

<211>50

<212>DNA

<213> combinatorial 7 BIP sequences

<400>31

acaatattca ggtagagatg ctggtgttta tctttctaat agcctgatga 50

<210>32

<211>22

<212>DNA

<213> combinatorial 8F 3 sequences

<400>32

tgattcatgt gatctgtact gt 22

<210>33

<211>25

<212>DNA

<213> combinatorial 8B 3 sequences

<400>33

actgttaaag ttactatgtt agagg 25

<210>34

<211>46

<212>DNA

<213> Combined 8 FIP sequences

<400>34

gtacattcca actcaagaac aacagacact tcaaaggaaa acaacc 46

<210>35

<211>46

<212>DNA

<213> Combined 8 BIP sequences

<400>35

ttcaggtaga gatgctggta cacctagttt atctttctaa tagcct 46

Claims (10)

Translated fromChinese

1.一种检测新型冠状病毒的LAMP引物组合，其特征在于，选自如SEQ ID NO:3～SEQ IDNO:6所示的引物组1、如SEQ ID NO:7～SEQ ID NO:10所示的引物组2、如SEQ ID NO:11～SEQ ID NO:14所示的引物组3、如SEQ ID NO:15～SEQ ID NO:18所示的引物组4、如SEQ IDNO:19～SEQ ID NO:22所示的引物组5、如SEQ ID NO:24～SEQ ID NO:27所示的引物组6、如SEQ ID NO:28～SEQ ID NO:31所示的引物组7、如SEQ ID NO:32～SEQ ID NO:35所示的引物组8中的任一种或几种。1. a LAMP primer combination for detecting novel coronavirus, is characterized in that, is selected from primer set 1 as shown in SEQ ID NO:3～SEQ ID NO:6, as shown in SEQ ID NO:7～SEQ ID NO:10 The primer set 2 of SEQ ID NO: 11～SEQ ID NO: 14, the primer set 3 as shown in SEQ ID NO: 15～SEQ ID NO: 18, the primer set 4 as shown in SEQ ID NO: 15～SEQ ID NO: 18, as shown in SEQ ID NO: 19～SEQ ID NO: 18 Primer set 5 shown in ID NO: 22, primer set 6 shown as SEQ ID NO: 24 to SEQ ID NO: 27, primer set 7 shown as SEQ ID NO: 28 to SEQ ID NO: 31, Any one or more of primer set 8 shown in SEQ ID NO:32 to SEQ ID NO:35.2.一种检测新型冠状病毒的LAMP试剂盒，其特征在于，包括如权利要求1所述的LAMP引物组合。2. A LAMP test kit for detecting novel coronavirus, characterized in that, comprising the combination of LAMP primers as claimed in claim 1.3.根据权利要求2所述的检测新型冠状病毒的LAMP试剂盒，其特征在于，所述引物的浓度为80-110μM。3. The LAMP kit for detecting novel coronavirus according to claim 2, wherein the concentration of the primer is 80-110 μM.4.根据权利要求3所述的检测新型冠状病毒的LAMP试剂盒，其特征在于，所述引物的浓度为100μM。4. The LAMP kit for detecting novel coronavirus according to claim 3, wherein the concentration of the primer is 100 μM.5.根据权利要求2所述的检测新型冠状病毒的LAMP试剂盒，其特征在于，所述LAMP试剂盒还包括反应缓冲液、BSst DNA聚合酶、SYBR/中性红染料、DNA模板和双蒸水。5. the LAMP test kit of detecting novel coronavirus according to claim 2, is characterized in that, described LAMP test kit also comprises reaction buffer, BSst DNA polymerase, SYBR/neutral red dye, DNA template and double distillation water.6.根据权利要求5所述的检测新型冠状病毒的LAMP试剂盒，其特征在于，所述反应缓冲液包括Tris-HCl、KCl、(NH₄)₂SO₄、MgSO₄和Tween-20。6. The LAMP kit for detecting novel coronavirus according to claim 5, wherein the reaction buffer comprises Tris-HCl, KCl, (NH₄ )₂ SO₄ , MgSO₄ and Tween-20.7.利用如权利要求2-6任一项所述LAMP试剂盒检测该新型冠状病毒的方法，其特征在于，包括如下步骤：7. utilize LAMP test kit as described in any one of claim 2-6 to detect the method for this novel coronavirus, it is characterised in that comprising the steps:步骤1，提取纯化待测样品的DNA；Step 1, extract and purify the DNA of the sample to be tested;步骤2，配制LAMP反应体系，进行环介导等温扩增反应；Step 2, prepare a LAMP reaction system, and perform a loop-mediated isothermal amplification reaction;步骤3，结果判断。Step 3, result judgment.8.根据权利要求7所述的检测新型冠状病毒的LAMP试剂盒，其特征在于，步骤2中的所述反应体系的体积为30μL，包括如下体积的试剂：反应缓冲液20μL、引物各0.05μL、BSstDNA聚合酶1μL、SYBR/中性红染料1μL、DNA模板4μL、余量为双蒸水。8. The LAMP kit for detecting novel coronavirus according to claim 7, wherein the volume of the reaction system in step 2 is 30 μL, including the reagents of the following volumes: 20 μL of reaction buffer, 0.05 μL of each primer , 1 μL of BSstDNA polymerase, 1 μL of SYBR/neutral red dye, 4 μL of DNA template, and the balance is double distilled water.9.根据权利要求7所述的检测新型冠状病毒的LAMP试剂盒，其特征在于，所述环介导等温扩增反应的条件为55～70℃，30～60min。9 . The LAMP kit for detecting novel coronavirus according to claim 7 , wherein the loop-mediated isothermal amplification reaction conditions are 55-70° C. for 30-60 min. 10 .10.根据权利要求7所述的检测新型冠状病毒的LAMP试剂盒，其特征在于，所述环介导等温扩增反应所采用的设备为普通PCR仪、实时PCR仪或恒温金属浴。10. the LAMP test kit of detecting novel coronavirus according to claim 7, is characterized in that, the equipment that described loop-mediated isothermal amplification reaction adopts is common PCR machine, real-time PCR machine or constant temperature metal bath.

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