技术领域technical field
本发明属于生物医学技术领域，具体涉及一种以HOXB9和PBX1作为生物标志物的胃癌检测试剂盒及应用。The invention belongs to the technical field of biomedicine, and in particular relates to a gastric cancer detection kit and application using HOXB9 and PBX1 as biomarkers.
背景技术Background technique
胃癌是消化道常见的恶性肿瘤，其发病率占全球每年新发肿瘤第四位，死亡率高居第二位。胃癌诊断和治疗方案的制定有赖于病理和分子病理诊断结果。分子病理可借助免疫组化检测技术，为胃癌的诊断、鉴别诊断、分型分期、化疗、靶向治疗和预后提供依据。免疫组化技术是利用标记的特异性抗体对组织细胞内肿瘤相关抗原的表达量和分布进行原位检测的技术。目前，已知有10多种胃癌相关标志物免疫组化指标应用于临床。Gastric cancer is a common malignant tumor of the digestive tract. Its incidence rate accounts for the fourth of new tumors every year in the world, and its mortality rate ranks the second. The formulation of diagnosis and treatment plan for gastric cancer depends on the results of pathological and molecular pathological diagnosis. Molecular pathology can provide evidence for the diagnosis, differential diagnosis, classification and staging, chemotherapy, targeted therapy and prognosis of gastric cancer with the help of immunohistochemical detection technology. Immunohistochemistry is a technique for in situ detection of the expression and distribution of tumor-associated antigens in tissue cells using labeled specific antibodies. At present, more than 10 immunohistochemical indicators related to gastric cancer markers are known to be used clinically.
由于胃癌的发病机制不明，现有的胃癌相关标志物临床判断胃癌的生长及预后效果欠佳。每个胃癌标志物检测需要至少一张切片，对组织消耗量大，无法判断标志物间彼此相关性，准确度不高。Due to the unknown pathogenesis of gastric cancer, the existing gastric cancer-related markers are not effective in clinically judging the growth and prognosis of gastric cancer. The detection of each gastric cancer marker requires at least one slice, which consumes a lot of tissue, and the correlation between markers cannot be judged, and the accuracy is not high.
发明内容Contents of the invention
为了克服现有技术中的缺陷，本发明的目的在于提供HOXB9和PBX1联合作为胃癌生物标志物在制备或筛选胃癌检测试剂中的用途。In order to overcome the defects in the prior art, the purpose of the present invention is to provide the combination of HOXB9 and PBX1 as gastric cancer biomarkers in the preparation or screening of gastric cancer detection reagents.
本发明的另一目的在于提供一种以HOXB9和PBX1联合作为生物标志物的胃癌检测试剂盒。Another object of the present invention is to provide a gastric cancer detection kit using the combination of HOXB9 and PBX1 as biomarkers.
本发明的另一目的在于提供一种检测胃癌的方法。Another object of the present invention is to provide a method for detecting gastric cancer.
本发明是通过以下技术方案实现的：The present invention is achieved through the following technical solutions:
本发明的第一方面，提供了HOXB9和PBX1联合作为胃癌生物标志物在制备或筛选胃癌检测试剂中的用途。The first aspect of the present invention provides the combination of HOXB9 and PBX1 as gastric cancer biomarkers in the preparation or screening of gastric cancer detection reagents.
优选地，所述HOXB9和PBX1联合作为胃癌组织生物标志物。Preferably, the combination of HOXB9 and PBX1 is used as a gastric cancer tissue biomarker.
优选地，所述胃癌检测试剂以HOXB9和PBX1联合作为检测对象。Preferably, the gastric cancer detection reagent uses the combination of HOXB9 and PBX1 as the detection object.
优选地，所述胃癌检测试剂中，含有HOXB9和PBX1联合作为标准品或阳性对照。Preferably, the gastric cancer detection reagent contains a combination of HOXB9 and PBX1 as a standard or a positive control.
所述标准品或阳性对照，用于检测样本组织中的HOXB9和PBX1含量。The standard or positive control is used to detect the contents of HOXB9 and PBX1 in sample tissues.
将HOXB9和PBX1联合作为胃癌生物标志物用于筛选胃癌检测试剂，是指将HOXB9与PBX1作为靶标应用于胃癌检测试剂的筛选。在一优选例中，基于所述的HOXB9与PBX1，筛选特异性识别HOXB9的试剂和特异性识别PBX1的试剂，从而作为检测胃癌的试剂，来检测样本组织中的HOXB9与PBX1水平。Combining HOXB9 and PBX1 as gastric cancer biomarkers for screening gastric cancer detection reagents refers to using HOXB9 and PBX1 as targets for screening gastric cancer detection reagents. In a preferred example, based on the HOXB9 and PBX1, a reagent that specifically recognizes HOXB9 and a reagent that specifically recognizes PBX1 are screened, so as to detect the levels of HOXB9 and PBX1 in the sample tissue as a reagent for detecting gastric cancer.
在一优选例中，基于所述的HOXB9与PBX1，筛选特异性识别HOXB9的抗体和特异性识别PBX1的抗体，从而作为从而作为检测胃癌的试剂。In a preferred example, based on the HOXB9 and PBX1, an antibody that specifically recognizes HOXB9 and an antibody that specifically recognizes PBX1 is screened to serve as a reagent for detecting gastric cancer.
优选地，所述胃癌检测试剂中，含有特异性识别HOXB9的试剂和特异性识别PBX1的试剂。Preferably, the gastric cancer detection reagent contains a reagent that specifically recognizes HOXB9 and a reagent that specifically recognizes PBX1.
优选地，所述胃癌检测试剂用于胃癌的判断、治疗方案的选择、和/或预后评估。Preferably, the gastric cancer detection reagent is used for the judgment of gastric cancer, the selection of treatment plan, and/or the assessment of prognosis.
本发明的第二方面，提供了特异性识别HOXB9的试剂和特异性识别PBX1的试剂在制备胃癌检测试剂盒中的用途。The second aspect of the present invention provides the use of the reagent specifically recognizing HOXB9 and the reagent specifically recognizing PBX1 in the preparation of a gastric cancer detection kit.
在一优选例中，特异性识别HOXB9的试剂选自特异性识别HOXB9的抗体。特异性识别PBX1的试剂选自特异性识别PBX1的抗体。In a preferred example, the reagent specifically recognizing HOXB9 is selected from antibodies specifically recognizing HOXB9. The reagent that specifically recognizes PBX1 is selected from antibodies that specifically recognize PBX1.
所述抗体包括单克隆抗体和多克隆抗体。The antibodies include monoclonal antibodies and polyclonal antibodies.
本发明的第三方面，提供一种胃癌检测试剂盒，包括特异性识别HOXB9的试剂和特异性识别PBX1的试剂。The third aspect of the present invention provides a gastric cancer detection kit, including a reagent that specifically recognizes HOXB9 and a reagent that specifically recognizes PBX1.
优选地，所述胃癌检测试剂盒以HOXB9和PBX1作为胃癌生物标志物。Preferably, the gastric cancer detection kit uses HOXB9 and PBX1 as gastric cancer biomarkers.
优选地，所述胃癌检测试剂盒以HOXB9和PBX1作为检测对象。Preferably, the gastric cancer detection kit uses HOXB9 and PBX1 as detection objects.
在一优选例中，特异性识别HOXB9的试剂选自特异性识别HOXB9的抗体。特异性识别PBX1的试剂选自特异性识别PBX1的抗体。In a preferred example, the reagent specifically recognizing HOXB9 is selected from antibodies specifically recognizing HOXB9. The reagent that specifically recognizes PBX1 is selected from antibodies that specifically recognize PBX1.
所述抗体包括单克隆抗体和多克隆抗体。The antibodies include monoclonal antibodies and polyclonal antibodies.
在一优选例中，特异性识别HOXB9的抗体包括用于检测HOXB9的第一抗体和用于检测所述第一抗体的第二抗体。特异性识别PBX1的抗体包括用于检测PBX1的第一抗体和和用于检测所述第一抗体的第二抗体。In a preferred example, the antibody specifically recognizing HOXB9 includes a first antibody for detecting HOXB9 and a second antibody for detecting the first antibody. Antibodies that specifically recognize PBX1 include a primary antibody for detecting PBX1 and a secondary antibody for detecting the primary antibody.
进一步地，所述第二抗体为荧光分子标记的第二抗体。Further, the second antibody is a fluorescent molecule-labeled second antibody.
在一优选例中，所述的试剂盒中还包括免疫荧光染色技术常用的试剂，免疫荧光染色技术常用的试剂包括：福尔马林固定液、石蜡、二甲苯、柠檬酸钠、封闭液中的一种或多种。In a preferred example, the kit also includes reagents commonly used in immunofluorescence staining techniques, which include: formalin fixative, paraffin, xylene, sodium citrate, blocking solution one or more of .
在一优选例中，所述试剂盒还包括阳性对照、阴性对照。所述阳性对照包括HOXB9和PBX1。所述阴性对照包括ddH2O。In a preferred example, the kit further includes a positive control and a negative control. The positive controls include HOXB9 and PBX1.The negative control included ddH2O.
优选地，所述胃癌检测试剂盒用于胃癌的判断、治疗方案的选择、和/或预后评估。Preferably, the gastric cancer detection kit is used for the judgment of gastric cancer, the selection of treatment plan, and/or the assessment of prognosis.
在本发明的第四方面，提供了一种检测胃癌的方法，包括检测样本组织中HOXB9和PBX1的水平。In the fourth aspect of the present invention, a method for detecting gastric cancer is provided, comprising detecting the levels of HOXB9 and PBX1 in sample tissue.
与现有技术相比，本发明的有益效果在于：Compared with prior art, the beneficial effect of the present invention is:
本发明基于发明人研究成果，通过检测两个新发现且彼此关联的胃癌标志物，更准确的判断胃癌的生长及预后；通过免疫荧光双染法，精确判断标志物间彼此关系，节约组织消耗量。HOXB9和PBX1同时作为新的胃癌标志物，可为胃癌的分子病理诊断提供依据。在胃癌组织细胞原位同时观察胃癌标志物HOXB9和PBX1的表达，易于观察评价两者之间的相关性。一张切片上同时检测两个相关的胃癌标志物，有助于节约标本用量，可应用于胃镜等取材较少的检测。Based on the research results of the inventor, the present invention can more accurately judge the growth and prognosis of gastric cancer by detecting two newly discovered and correlated gastric cancer markers; through the immunofluorescent double staining method, the relationship between the markers can be accurately judged to save tissue consumption quantity. HOXB9 and PBX1 are simultaneously new markers of gastric cancer, which can provide evidence for molecular pathological diagnosis of gastric cancer. The expression of gastric cancer markers HOXB9 and PBX1 was simultaneously observed in situ in gastric cancer tissue cells, which is easy to observe and evaluate the correlation between the two. Simultaneous detection of two related gastric cancer markers on one slice helps to save the amount of specimens, and can be applied to the detection of less samples such as gastroscopy.
采用本发明的检测试剂盒对组织样本中的HOXB9和PBX1进行联合检测，从而诊断和检测胃癌，检测结果灵敏度好，特异性高。可有效用于胃癌的早期筛查和/或预后评估，具有良好的临床应用前景。The detection kit of the invention is used to jointly detect HOXB9 and PBX1 in tissue samples, thereby diagnosing and detecting gastric cancer, and the detection result has good sensitivity and high specificity. It can be effectively used for early screening and/or prognosis evaluation of gastric cancer, and has good clinical application prospects.
附图说明Description of drawings
图1：左图表示HOXB9在正常组织中高表达，右图表示HOXB9在胃癌组织中低表达。Figure 1: The left panel shows the high expression of HOXB9 in normal tissues, and the right panel shows the low expression of HOXB9 in gastric cancer tissues.
图2：左图表示PBX1在正常组织中低表达，右图表示PBX1在胃癌组织中高表达。Figure 2: The left figure shows the low expression of PBX1 in normal tissues, and the right figure shows the high expression of PBX1 in gastric cancer tissues.
图3：HOXB9和PBX1的表达在胃癌组织中的相关性。Figure 3: The correlation between the expression of HOXB9 and PBX1 in gastric cancer tissues.
具体实施方式Detailed ways
以下通过特定的具体实例说明本发明的实施方式，本领域技术人员可由本说明书所揭露的内容轻易地了解本发明的其它优点与功效。本发明还可以通过另外不同的具体实施方式加以实施或应用，本说明书中的各项细节也可以基于不同观点与应用，在没有背离本发明的精神下进行各种修饰或改变。Embodiments of the present invention are described below through specific examples, and those skilled in the art can easily understand other advantages and effects of the present invention from the contents disclosed in this specification. The present invention can also be implemented or applied through other different specific implementation modes, and various modifications or changes can be made to the details in this specification based on different viewpoints and applications without departing from the spirit of the present invention.
在进一步描述本发明具体实施方式之前，应理解，本发明的保护范围不局限于下述特定的具体实施方案；还应当理解，本发明实施例中使用的术语是为了描述特定的具体实施方案，而不是为了限制本发明的保护范围；在本发明说明书和权利要求书中，除非文中另外明确指出，单数形式“一个”、“一”和“这个”包括复数形式。Before further describing the specific embodiments of the present invention, it should be understood that the protection scope of the present invention is not limited to the following specific specific embodiments; it should also be understood that the terms used in the examples of the present invention are to describe specific specific embodiments, It is not intended to limit the protection scope of the present invention; in the description and claims of the present invention, unless the context clearly indicates otherwise, the singular forms "a", "an" and "the" include plural forms.
当实施例给出数值范围时，应理解，除非本发明另有说明，每个数值范围的两个端点以及两个端点之间任何一个数值均可选用。除非另外定义，本发明中使用的所有技术和科学术语与本技术领域技术人员通常理解的意义相同。除实施例中使用的具体方法、设备、材料外，根据本技术领域的技术人员对现有技术的掌握及本发明的记载，还可以使用与本发明实施例中所述的方法、设备、材料相似或等同的现有技术的任何方法、设备和材料来实现本发明。When the examples give numerical ranges, it should be understood that, unless otherwise stated in the present invention, the two endpoints of each numerical range and any value between the two endpoints can be selected. Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art. In addition to the specific methods, equipment, and materials used in the embodiments, according to those skilled in the art's grasp of the prior art and the description of the present invention, the methods, equipment, and materials described in the embodiments of the present invention can also be used Any methods, apparatus and materials of the prior art similar or equivalent to the practice of the present invention.
除非另外说明，本发明中所公开的实验方法、检测方法、制备方法均采用本技术领域常规的分子生物学、生物化学、染色质结构和分析、分析化学、细胞培养、重组DNA技术及相关领域的常规技术。这些技术在现有文献中已有完善说明，具体可参见Sambrook等MOLECμLAR CLONING：A LABORATORY MANUAL，Second edition，Cold Spring Harbor LaboratoryPress，1989and Third edition，2001；Ausubel等，CURRENT PROTOCOLS IN MOLECμLARBIOLOGY，John Wiley&Sons，New York，1987and periodic updates；the series METHODSIN ENZYMOLOGY，Academic Press，San Diego；Wolffe，CHROMATIN STRUCTURE ANDFUNCTION，Third edition，Academic Press，San Diego，1998；METHODS IN ENZYMOLOGY，Vol.304，Chromatin(P.M.Wassarman and A.P.Wolffe，eds.)，Academic Press，SanDiego，1999；和METHODS IN MOLECμLAR BIOLOGY，Vol.119，Chromatin Protocols(P.B.Becker，ed.)Humana Press，Totowa，1999等。Unless otherwise stated, the experimental methods, detection methods, and preparation methods disclosed in the present invention all adopt conventional molecular biology, biochemistry, chromatin structure and analysis, analytical chemistry, cell culture, recombinant DNA technology and related fields in the technical field conventional technology. These techniques have been fully described in the existing literature. For details, see Sambrook et al. MOLEC μ LAR CLONING: A LABORATORY MANUAL, Second edition, Cold Spring Harbor Laboratory Press, 1989 and Third edition, 2001; Ausubel et al., CURRENT PROTOCOLS IN MOLEC μ LAR BIOLOGY, John Wiley & Sons, New York, 1987 and periodic updates; the series METHODSIN ENZYMOLOGY, Academic Press, San Diego; Wolffe, CHROMATIN STRUCTURE ANDFUNCTION, Third edition, Academic Press, San Diego, 1998; METHODS IN ENZYMOLOGY, Vol.304, Chromatinass (Plf.Wo.M.W. eds.), Academic Press, San Diego, 1999; and METHODS IN MOLEC μLAR BIOLOGY, Vol. 119, Chromatin Protocols (P.B. Becker, ed.) Humana Press, Totowa, 1999, etc.
术语、缩略语terms, abbreviations
qRT-PCR：quantitative real-time polymerase chain reaction，实时荧光定量PCRqRT-PCR: quantitative real-time polymerase chain reaction, real-time fluorescent quantitative PCR
灵敏度：即真阳性率，真阳性与真阳性+假阴性的比例。Sensitivity: the true positive rate, the ratio of true positives to true positives + false negatives.
特异性：即真阴性率，真阴性与真阴性+假阳性的比例。Specificity: the true negative rate, the ratio of true negatives to true negatives + false positives.
实施例1.通过免疫荧光双染法对胃癌组织内HOXB9和PBX1的表达水平和部位进行检测Example 1. Detection of the expression level and location of HOXB9 and PBX1 in gastric cancer tissue by immunofluorescence double staining
一、本实施例通过免疫荧光双染法对胃癌组织内HOXB9和PBX1的表达水平和部位进行检测，步骤如下：1. In this example, the expression level and location of HOXB9 and PBX1 in gastric cancer tissue were detected by immunofluorescence double staining, and the steps were as follows:
(1)胃镜取材或手术切除胃癌组织，常规10％福尔马林固定，石蜡包埋，5mm切片。(1) Gastric cancer tissues were collected by endoscopy or surgically removed, fixed in 10% formalin, embedded in paraffin, and sectioned in 5 mm.
(2)烤片：60摄氏度，1小时。(2) Baked slices: 60 degrees Celsius, 1 hour.
(3)脱腊：二甲苯溶液I中15分钟，二甲苯溶液II中15分钟，无水乙醇I中5分钟，无水乙醇II中5分钟，90％乙醇I中5分钟，90％乙醇II中5分钟，70％乙醇中5分钟，双蒸水洗2次。(3) Dewaxing: 15 minutes in xylene solution I, 15 minutes in xylene solution II, 5 minutes in absolute ethanol I, 5 minutes in absolute ethanol II, 5 minutes in 90% ethanol I, 90% ethanol II 5 minutes in 70% ethanol, 5 minutes in double distilled water, and washed twice.
(4)抗原修复：高压修复法，加入0.01mol/L柠檬酸钠(Ph 6.0)，加热10分钟后缓慢冷却至室温。(4) Antigen retrieval: high-pressure retrieval method, adding 0.01mol/L sodium citrate (Ph 6.0), heating for 10 minutes and then slowly cooling to room temperature.
(5)封闭：加10％正常驴血清，室温孵育封闭30分钟。(5) Blocking: add 10% normal donkey serum, incubate at room temperature for 30 minutes to block.
(6)加一抗：滴加含小鼠抗HOXB9和大鼠抗PBX1的一抗抗体，4度孵育过夜，PBS震荡洗涤3遍。(6) Add primary antibody: add primary antibody containing mouse anti-HOXB9 and rat anti-PBX1 dropwise, incubate overnight at 4°C, wash with PBS shaking 3 times.
(7)加二抗：滴加FITC和APC标记的兔血清来源的二抗，室温孵育1小时，PBS震荡洗涤3遍。(7) Add secondary antibody: add FITC- and APC-labeled secondary antibody derived from rabbit serum dropwise, incubate at room temperature for 1 hour, wash with PBS shaking 3 times.
(8)将盖玻片放于纸巾上，滴加Gelvatol一滴于盖玻片中央。翻过载玻片于盖玻片上，用铝箔避光放置30分钟。(8) Put the coverslip on a paper towel, and add one drop of Gelvatol to the center of the coverslip. Turn over the slide on the coverslip and place it in the dark for 30 minutes with aluminum foil.
(9)显微镜下观察。(9) Observe under a microscope.
二、本实施例首次发现：在胃癌组织中，HOXB9和PBX1的表达水平与胃癌生长和淋巴转移密切相关；HOXB9和PBX1之间有负相关性：2. This example found for the first time that in gastric cancer tissues, the expression levels of HOXB9 and PBX1 are closely related to gastric cancer growth and lymphatic metastasis; there is a negative correlation between HOXB9 and PBX1:
(1)如表1和图1所示，HOXB9在正常胃粘膜组织中表达较高，而胃癌组织中表达水平降低。(1) As shown in Table 1 and Figure 1, HOXB9 is highly expressed in normal gastric mucosal tissues, while its expression level is decreased in gastric cancer tissues.
表1：HOXB9表达水平降低可导致胃癌生长及淋巴转移密切Table 1: Decreased expression of HOXB9 can lead to close relationship between gastric cancer growth and lymphatic metastasis
(2)如图2和表2所示，PBX1在正常胃粘膜组织中表达较低，而胃癌组织中表达水平升高：(2) As shown in Figure 2 and Table 2, the expression level of PBX1 in normal gastric mucosal tissue is low, while the expression level in gastric cancer tissue is increased:
表2：PBX1表达水平升高可导致胃癌生长及淋巴转移密切Table 2: Increased expression of PBX1 can lead to gastric cancer growth and lymphatic metastasis
(3)如图3所示，HOXB9和PBX1的表达在胃癌组织中的相关性：HOXB9表达降低时，PBX1的表达水平会升高，两者之间为负相关性。(3) As shown in Figure 3, the correlation between the expression of HOXB9 and PBX1 in gastric cancer tissues: when the expression of HOXB9 decreases, the expression level of PBX1 will increase, and there is a negative correlation between the two.
实施例2.以HOXB9和PBX1作为生物标志物的胃癌检测试剂盒的构建及应用Example 2. Construction and application of a gastric cancer detection kit using HOXB9 and PBX1 as biomarkers
本实施例构建以HOXB9和PBX1作为生物标志物的胃癌检测试剂盒，所述试剂盒包括：用于检测HOXB9的试剂和用于检测PBX1的试剂。用于检测HOXB9的试剂包括用于检测HOXB9的第一抗体和用于检测所述第一抗体的第二抗体。用于检测PBX1的试剂包括用于检测PBX1的第一抗体和用于检测所述第一抗体的第二抗体。In this example, a gastric cancer detection kit using HOXB9 and PBX1 as biomarkers was constructed, and the kit included: a reagent for detecting HOXB9 and a reagent for detecting PBX1. A reagent for detecting HOXB9 includes a primary antibody for detecting HOXB9 and a secondary antibody for detecting the primary antibody. A reagent for detecting PBX1 includes a primary antibody for detecting PBX1 and a secondary antibody for detecting the primary antibody.
具体地，本实施例的胃癌检测试剂盒通过免疫荧光双染法对胃癌组织内HOXB9和PBX1的表达水平进行检测。用于检测HOXB9的第一抗体采用小鼠源HOXB9抗体，用于检测所述第一抗体的第二抗体采用FITC标记的兔源抗体。用于检测PBX1的第一抗体采用大鼠源PBX1抗体，用于检测所述第一抗体的第二抗体采用APC标记的兔源抗体。Specifically, the gastric cancer detection kit of this embodiment detects the expression levels of HOXB9 and PBX1 in gastric cancer tissue by immunofluorescence double staining. The primary antibody used to detect HOXB9 was a mouse-derived HOXB9 antibody, and the second antibody used to detect the first antibody was a FITC-labeled rabbit-derived antibody. The primary antibody used to detect PBX1 is a rat-derived PBX1 antibody, and the second antibody used to detect the first antibody is an APC-labeled rabbit-derived antibody.
采用构建好的以HOXB9和PBX1作为生物标志物的胃癌检测试剂盒，参照实施例1的方法，检测100个临床样本，以判断是否为胃癌样本以及是否有淋巴转移。Using the constructed gastric cancer detection kit using HOXB9 and PBX1 as biomarkers, referring to the method in Example 1, 100 clinical samples were detected to determine whether they were gastric cancer samples and whether there was lymphatic metastasis.
以样本中：HOXB9表达量和PBX1表达量的比值作为诊断指标，当HOXB9表达量和PBX1表达量的比值<1(荧光信号以PBX1占主导)算为胃癌阳性样本；当HOXB9表达量和PBX1表达量的比值>1(荧光信号以HOXB9占主导)作为胃癌阴性样本。In the sample, the ratio of HOXB9 expression to PBX1 expression was used as a diagnostic index. When the ratio of HOXB9 expression to PBX1 expression was <1 (the fluorescent signal was dominated by PBX1), it was regarded as a positive gastric cancer sample; when the ratio of HOXB9 expression to PBX1 expression The ratio of the amount > 1 (the fluorescent signal is dominated by HOXB9) was regarded as a gastric cancer negative sample.
其中，43个样本算为胃癌阳性样本；57个样本算为阴性样本。Among them, 43 samples were counted as gastric cancer positive samples; 57 samples were counted as negative samples.
本实例首次发现，在胃癌组织中同时检测HOXB9和PBX1，并以HOXB9表达量和PBX1表达量的比值作为诊断指标，在没有明显降低灵敏度的同时显著增加了特异性，见表3：This example found for the first time that simultaneous detection of HOXB9 and PBX1 in gastric cancer tissue, and using the ratio of HOXB9 expression to PBX1 expression as a diagnostic index, significantly increased specificity without significantly reducing sensitivity, as shown in Table 3:
表3：HOXB9和PBX1三种检测方式灵敏度和特异性的比较Table 3: Comparison of the sensitivity and specificity of the three detection methods of HOXB9 and PBX1
综上所述，采用本发明的检测试剂盒对组织样本中的HOXB9和PBX1进行联合检测，从而诊断和检测胃癌，检测结果灵敏度好，特异性高。可有效用于胃癌的早期筛查和/或预后评估，具有良好的临床应用前景。In summary, the detection kit of the present invention is used to jointly detect HOXB9 and PBX1 in tissue samples, thereby diagnosing and detecting gastric cancer, and the detection result has good sensitivity and high specificity. It can be effectively used for early screening and/or prognosis evaluation of gastric cancer, and has good clinical application prospects.
以上的实施例是为了说明本发明公开的实施方案，并不能理解为对本发明的限制。此外，本文所列出的各种修改以及发明中方法、组合物的变化，在不脱离本发明的范围和精神的前提下对本领域内的技术人员来说是显而易见的。虽然已结合本发明的多种具体优选实施例对本发明进行了具体的描述，但应当理解，本发明不应仅限于这些具体实施例。事实上，各种如上所述的对本领域内的技术人员来说显而易见的修改来获取发明都应包括在本发明的范围内。The above examples are intended to illustrate the disclosed embodiments of the present invention, and should not be construed as limiting the present invention. In addition, various modifications set forth herein, as well as changes in the method and composition of the invention, will be apparent to those skilled in the art without departing from the scope and spirit of the invention. Although the invention has been specifically described in connection with various specific preferred embodiments of the invention, it should be understood that the invention should not be limited to these specific embodiments. In fact, various modifications as mentioned above which are obvious to those skilled in the art to obtain the invention should be included in the scope of the present invention.
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| CN201610662720.3ACN106124764B (en) | 2016-08-12 | 2016-08-12 | Stomach cancer detection kit and application using HOXB9 and PBX1 as biomarker |
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| CN201610662720.3ACN106124764B (en) | 2016-08-12 | 2016-08-12 | Stomach cancer detection kit and application using HOXB9 and PBX1 as biomarker |
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| CN111518893A (en)* | 2020-05-11 | 2020-08-11 | 深圳市人民医院 | Uremia markers and their applications |
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