CN85107855A - The goods that are used for diagnostic check - Google Patents

Abstract

The present invention introduced a kind of in diagnostic check useful articles, be used for soluble reagents in the used measured quantity of diagnostic check dispensing.It is the porous body of a moulding, and soluble reagents is dispersed in the hole of this porous body.These article are distributed in the hole of porous body this soluble reagents by the porous body that soluble reagents is added moulding with the solution form to prepare.Then, dry this formed body for example, passes through lyophilization.Injection will be dissolved this reagent again by the solvent of porous body.In the embodiment an of the best, this reagent be a kind of in immunity inspection the monoclonal antibody of useful mark.

Description

The goods that are used for diagnostic check

The present invention generally speaking is diagnostic check method field, and relevant with used device and article in these diagnostic check methods.On the other hand, the invention relates to the field of the reagent of diagnostic check method.Moreover the present invention also is about being used for accurately sending a kind of device of a small amount of soluble reagents.

Super-sensitive several diagnostic check method, several coordination acceptor methods of inspection of particularly several immunity inspection methods and other can accurately be measured the seldom quantitative analysis thing that is present in the body fluid sample in for example serum, blood plasma, urine and the saliva.Based on the accurate measurement of small quantity of reagent and the fact of transfer, these analyses are complicated.For example, many immunity inspection methods are used a kind of soluble antigen or antibody.In these methods of inspection, usually provide this antigen or antibody is dissolved and shift with transfer pipet with the form of lyophilization.

In the coordination acceptor check of immunity inspection or other complex characteristics, other problem is owing to used the sample that comprises a large amount of undesirable cell fragments when collecting to cause.The formation that these cell fragments suppress or sheltered the Ag-Ab complex compound.These cell fragments also can block or block some stages of testing and handling that enter into.

When existing diagnostic tool is when being used by trained personnel, these complicated problems can overcome.If yet do not get rid of, in many cases, the possibility by the personnel that lack this training finish check reliably also will significantly reduce.Concerning inexperienced personnel, to plan under the situation with the product of selling to the rawness personnel on the sales counter, this is the most serious problem.So the device of eliminating these problems will make these methods of inspection convenient to the trainer of medium level.

The present invention is a kind of article or the device that can reduce or eliminate above-mentioned these problems.These article (device) comprise the porous body of a moulding, it contain measured quantity full dispersion but adhere to and coated the surperficial of hole or have a kind of soluble reagents in the hole in one's care.In the porous body of moulding, a kind of solvent that adds this reagent reconstitutes the solution of this soluble reagents effectively, so that it can use in check.By the size of selecting hole, these article (device) can be used as prefilter in diagnostic check, and the size in hole should be small enough to filter out undesirable cell fragment.And, because joining the amount of the soluble reagents of moulding porous body can pre-determine and accurately introduce, user subsequently only need handle this moulding porous body and go so that the soluble reagents of measured quantity is joined in the diagnostic check, the necessity that this has just eliminated preparation and has inhaled solution with suction pipe.As hereinafter introducing in detail, to use in a kind of monoclonal antibody or the immunity inspection method of polyclonal antibody preparation of solubility at some as reagent, this article (device) have special effectiveness.At last, this porous body can prepare the reaction chamber of the reaction product between soluble reagents and analyte or other component.

Therefore, the objective of the invention is to make the simplified control of diagnostic check method.

Another object of the present invention provide a kind of with an energy in a process, the simple device of soluble reagents in the diagnostic check particularly.

Another object of the present invention provides one can be used as the dispensing devices that filtration unit can be used as the soluble reagents of the measured quantity in advance in this check simultaneously again in diagnostic check.

According to the optimum implementation that describes below, the mode that realizes these purposes will be tangible.

As mentioned above, the invention provides the article (device) that the diagnostic check method of using soluble reagents is had special applications.The check of coordination acceptor is arranged in some are checked so usually, include, but are not limited to soluble antigen as soluble reagents, or with polyclone or monoclonal antibody formulation as the immunity inspection method of soluble reagents with the DNA probe checkout method of nucleic acid oligomer as soluble reagents.The present invention be particularly suitable for a kind of solubility labelled antibody, preferably monoclonal antibody as the check component method of inspection.Therefore, obviously the those of skill in the art in the technology of the present invention field know that the present invention can be used in combination with other soluble reagents, and still, the present invention will be according to introducing as soluble reagents with monoclonal antibody, so this application of the present invention only is exemplary.

Article of the present invention (device) include one contain measured quantity be dispersed in the porous structure entirely, coated or stick to the single hole surface or have the formed body of the soluble reagents in porous body in one's care.The given shape of porous body is not strict the qualification.And always having a kind of specialized application usually is shape easily.The most common situation is that the shape of porous body is suitable for placing in the vessel or is fit to admit it to do in another container of intended purpose.With reference to figure 1, porous body is shown in cylinder 30.Cylinder 30 will have the diameter of tight insertion bulge.Though the same of other shape also has, for example cone or cube may be more suitable to some application.

The most a kind of polymeric material of porous body.For example resemble polystyrene, tygon, nylon, polyester and cellulose acetate etc.Glass or inorganic fibre formed body also can use.Certainly the material of actual selection must be not react with soluble reagents, and does not combine and damage subsequently the ability that this reagent dissolves again with any alternate manner with soluble reagents.The physical arrangement of formed body is unimportant, and for example, it can obtain by organized single fiber or by discharging frothing agent generates opening when it cools off single foam in the molten mass again.Certainly, the technology of making this formed body is well-known, nor is ingredient of the present invention.Below will be clearer, being used to select the main standard of specific formed body will be to consider the size in hole and the intermiscibility of porous body and soluble reagents.

Soluble reagents add in the moulding porous body be by the solution with it join in the formed body and carry out drying process subsequently, at thermally sensitive biological products for example under the situation of antigen, antibody or nucleic acid low polymer, preferably lyophilization is operated and is finished.Usually adopt to be less than and make type body saturated solution amount fully.If do not do like this, coated is at the outside surface of formed body probably for most reagent, and it may easily be removed by friction.For example, if the saturated volume of formed body is 250 μ l, wish to form under the situation of reagent membrane at outside surface avoiding, most solutions is moved to outside surface and is formed local saturated mode, adds reagent in total solution of about 200 μ l.

In addition, before or after adding soluble reagents, in storing or disposing, formed body preferably all is placed in the container of protection.Under columnar situation, can use a protection sleeve.This protection container can easily receive on another structure body that is used in the check or from it face take off.

A kind ofly be used to carry out the particularly optimum device of immunity inspection method of diagnostic check method, can utilize article of the present invention (device) with it, this device is introduced in the application number in 11 days Mays in 1984 of people such as Valkirs is 609395 United States Patent (USP) " method and apparatus of immunity inspection ", the content of the disclosure is incorporated among the present invention by quoting here, and this installs as shown in Figure 2.

Therefore, in Fig. 2, shown a bulge 10, though it can be any other suitable shape.Container 10 has only a upper end open 12 that is made of outer wall 14.This container can be made with glass or suitable plastics, and as shown in Figure 2, container 10 also has alower openings 16, and 16 insert movablychock plug 18 and insert to allowporous member 20 in this opening.Container 10 also has an annular flake or filtrator disk and parts of the confession selecting foruse 21, and the effect of these parts will be introduced below.These parts are placed on thecylindrical parts 22, and this absorption piece also throughport 16 inserts.

The part of container 10 (referring to label 24) is shrunk to as shown in Figure 2, makes sample toparts 20 so that a funnel to be provided, and guarantees sample and other condiment realization cleaning effectively onparts 20.

Therefore, this part volume of the following container of the size ofparts 22 and constriction 10 is preferably selected like this, so that make the whole liquid that add this device in the once check receive and to be retained in the absorption piece 22.Exhaust apparatus (not illustrating in Fig. 2), for example aperture is arranged near the bottom of container 10, with the air of overflowing and will discharge.In addition, the bottom of container 10 can be removed, and makes liquid pass throughparts 20 and 22 by the bottom amount discharge container.Yet, because this article (device) plan arbitrarily to use, and plan with simply and the hygiene method to make things convenient for the disposal of sample, the structure shown in the most handy Fig. 2.

Parts 20 are used for filtering out cellulose and other material from sample, and the carrier that perhaps for example is used as a kind of binding antibody that is verified antigen is so that remove this antigen from solution.Under any circumstance, solution example through port 12 introducings to be analyzed are added on the parts 20.Infiltrate afterparts 20 and downward liquid flow enterabsorption piece 22 at sample, under the situation of " sandwich method of inspection ", labelled antibody solution (preferably monoclonal antibody) is added on theparts 20.

Then, labelled antibody or combine with the antigen that is bound by on the antibody on theparts 20, or be captured inparts 20 lip-deep cellulosic materials and combine.Ifparts 20 have monoclonal antibody constraint thereon, and labelled antibody also is a kind of monoclonal antibody, these two kinds of antibody are selected to be attached to the application on June 6th, 323498,1981 of United States Patent (USP) 4376110(application number) on the binding site of the non-interference antigen described.Wherein disclosed content with reference to and be incorporated among the present invention.

The most handy enzyme of soluble antibody makes marks, though the immune marker of other routine suitably also can use under the situation, for example can be with fluorescent marker or radioisotope labeling thing.

By after theparts 20, washing lotion addsparts 20 not have the labelled antibody of combination and to enterparts 22 in the flushingparts 20 at labelled antibody solution.The incline structure ofwall 24 provides a monoblock type funnel to be beneficial to washing lotion is added on the adhesion residue that wall gets on to remove labelled antibody solution.

Before being added to labelled antibody solution and cleaning fluid on theparts 20, have to allow be captured inparts 20 on or antibody solution inparts 20 spaces in or antigen more widely combination the latent period of a weak point earlier, thereby proposed the sensitivity of check.Yet this latent period or do not need or very short, promptly it was at 60 seconds orders of magnitude or shorter.It is faster when the flow of solution that contains antigen or marked body byparts 20 is directed at a ratio and does not have flow of solution in conjunction with speed.

If the antibody labeling thing is a kind of enzyme, afterparts 20 supernatant eccysis go there is not the antibody of combination, the enzyme matrix solution joins on theparts 20, if target antigen or with the antibodies that is bound by on theparts 20, or combine with cellulose on theparts 20, target antigen will combine with a part of labelled antibody so.Enzyme will cause that matrix reacts, if suitably select, and can produce one can use by eye or by instrument detecting to change color.

Using under the situation of cellulose acetate material,absorption piece 22 can non-ly combine with labelled antibody on its upper surface specifically.Therefore, some are just occurred on this surface under theparts 20 by the visible change color of eye.For avoiding this visible change color to extend through on theparts 20, be preferably in the polystyrene of placing porous betweenparts 20 and 22 or other and the separating component (in Fig. 2, representing) can be non-made with the material of antibodies specifically withlabel 21.

Fig. 3 represents to have the use of the present invention of Fig. 2 article (device).Therefore, the formed body among Fig. 1 30 is set in the space that thelimit wall 14 by container 10 limits.For its effect is described, with describe the polymeric article of using among Fig. 3 (device) carry out human chorionic gonadotrophin, HCG, a kind of period of gestation raise also can be in urine detected hormone checking method.In this case, be monoclonal antibody to the HCG soluble reagents at 30 li lyophilizations of parts, and parts itself are made with cellulose.This monoclonal antibody is with the alkaline phosphate mark of enzyme.Though other enzyme labeling thing, fluorescent marker or even radioactive label also all be operable.

The top that the urine samples of measured quantity is added to mouldingporous body 30 is preferably with calculating good sample size that can complete saturated porous body.Because sample size can saturated porous body,, and play a lytic antibody or a big at least antibody so sample just is retained in the hole of porous body.By the size of suitable selecting hole, this porous body also can be used as a prefilter, separates the epithelial cell that exists in urine of possibility plugmembers 20.

Urine samples allows to be retained in and is long enough to lytic antibody in theparts 30 and it can be combined with all HCG in the sample.Therefore, the immune response ofparts 30 antagonists and HCG plays a reactor.

After such latent period, add cleaning fluid, it can be other urine so that wash out this antibody-antigen complex compound fromparts 30 supernatants.

When this antibody-antigen complex compound washed out fromparts 30, its contact was also passed through diaphragm 20.Take off and abandonparts 30 then, this is just for to be bound by the antibody of solid phase and combining of HCG: the labelled antibody complex compound provides chance and it has been removed from solution.Unconjugated labelled antibody is cleaned throughparts 20 and enters absorber 22.

At sample and washings by after theparts 20, a kind of a spot of alkaline phosphate that contains for example goes and is strapped in enzyme reaction on the filtrator in the phosphatic matrix solution of the indoxyl top that is added toparts 20, consequently HCG reacts with being strapped in the antibody on theparts 20 and the soluble antibody of enzyme labeling, the indication that to form visible blue look be this reaction.

When recommending a this step to connect single step reaction, in some cases, can use urine capacity above the porous member saturated volume.Therefore, soluble antibody in sample and unfiltered material are directly imported parts 20.Certainly, the reaction time between the HCG is to have reduced in antibody and the sample.Therefore, a this step connects the step in a step and usually provides a kind of sensitiveer method of inspection.

In another kind was used, moulding porous body here bread contained a kind of antibody, and it is the antibody at the antigen on a kind of cellulose grain surface.In this case, the size inparts 30 holes must be selected like this, and the object that will be trapped inparts 20 surfaces after making can pass through.Yet the size inparts 30 holes can be selected to such an extent that make may plugmembers 20 and the maxicell of interference test must not pass through.This program is particularly suitable for the check of bacterium, fungi, bacterial parasite and other special biosome, and the detection of the virus that can filter out and corresponding antigens is arranged.

In another one was used, this moulding porous body just was used for sending easily the soluble antibody of measured quantity.Like this, after sample has been added toparts 20, withparts 30 patchholes 12 and for example a kind of antibody of the solvent of this reagent, allows to flow and go to dissolve and send the surface of antibody toparts 20 byparts 30.

Introducing above only is the example explanation of article purposes of the present invention, and therefore, scope of the present invention should consider only to be subjected to the restriction of claims.

Claims (21)

1, a kind of diagnostic check article that utilize the measured quantity soluble reagents, it comprises that a soluble reagents is dispersed in the moulding porous body in the moulding body opening, by allowing a kind of antibody solvent by the moulding porous body, this soluble reagents of elution from the porous body.

2, according to the article of claim 1, wherein soluble reagents is a lyophilization in the hole of moulding porous body.

3, according to the article of claim 1 or 2, diagnostic check therein is the check of coordination acceptor, and soluble reagents that to be above-mentioned coordination acceptor right is a kind of.

4, according to the goods of claim 3, wherein soluble reagents is to select from polyclonal antibody, monoclonal antibody, antigen or nucleic acid polymer preparation.

5, according to the goods of claim 4, wherein soluble reagents is a kind of monoclonal antibody.

6, according to the goods of claim 1 or 2, wherein the moulding porous body is the object of a kind of polymeric material, glass or inorganic fibre.

7, according to the goods of claim 6, wherein polymeric material is to select from nylon, polyester, polyolefin, polystyrene or acetate fiber.

8, according to the article of claim 7, wherein soluble reagents is to select from polyclonal antibody, monoclonal antibody, antigen or nucleic acid oligomer preparation.

9, article according to Claim 8, wherein soluble reagents is a kind of monoclonal antibody.

10, in a sample diagnostic check, use the soluble reagents of measured quantity, improvements comprise by allowing a solvent of this reagent be dispersed in moulding porous body in the hole by a soluble reagents, thereby soluble reagents is dissolved in this solvent, and wash out, and this soluble reagents is added in this check from this formed body.

11, according to the method for inspection of claim 10, soluble reagents is to select from polyclonal antibody preparation, monoclonal antibody, antigen or nucleic acid oligomer therein.

12, according to the method for inspection of claim 10 or 11, wherein soluble reagents is labeled, so that can check.

13, according to the method for inspection of claim 12, wherein this label is to select from enzyme, fluorescent material or radioactive nuclide.

14, according to the method for inspection of claim 10 or 11, wherein the moulding porous body is that polymeric material, glass or inorganic fibre are made.

15, according to the method for inspection of claim 14, wherein polymeric material is to select from nylon, polyolefin, polyester, polystyrene or acetate fiber.

16, according to the diagnostic check method of claim 10 or 11, wherein the moulding porous body is used for from sample filter fiber material.

17, according to the diagnostic check method of claim 10 or 11, sample is to select from urine, serum, blood plasma or saliva therein.

18, according to the diagnostic check method of claim 16, sample is to select from urine, serum, blood plasma or saliva therein.

19, according to the diagnostic check method of claim 10 or 11, wherein solvent is a sample to be tested.

20, according to the diagnostic check method of claim 19, sample is a urine therein.

21, according to the diagnostic check method of claim 19, soluble reagents is reacted with sample component in the hole of the hole body of porous body therein.

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