WO2023273623A1 - 检测血浆中piR-hsa-120522的试剂在制备肺癌筛查或诊断试剂盒中的用途 - Google Patents

检测血浆中piR-hsa-120522的试剂在制备肺癌筛查或诊断试剂盒中的用途 Download PDF

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WO2023273623A1
WO2023273623A1 PCT/CN2022/091993 CN2022091993W WO2023273623A1 WO 2023273623 A1 WO2023273623 A1 WO 2023273623A1 CN 2022091993 W CN2022091993 W CN 2022091993W WO 2023273623 A1 WO2023273623 A1 WO 2023273623A1
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lung cancer
reagent
pir
hsa
plasma
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董静思
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四川大学华西医院
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    • C12Q2600/00—Oligonucleotides characterized by their use
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  • the invention belongs to the field of in vitro diagnostic reagents, and specifically relates to the use of a reagent for detecting piR-hsa-120522 in blood plasma in the preparation of a screening or diagnostic kit for lung cancer and/or its precancerous lesions.
  • Lung cancer is one of the most common malignant tumors, and the mortality rate of lung cancer ranks first among malignant tumors worldwide. Lung cancer is one of the malignant tumors with the fastest increasing morbidity and mortality and the greatest threat to the health and life of the population. In the past 50 years, many countries have reported that the morbidity and mortality of lung cancer have increased significantly. The incidence and mortality of lung cancer in men rank first among all malignant tumors, and the incidence and mortality of lung cancer occupy the second place in women. The pathogenesis of lung cancer has not yet been fully elucidated. A large number of research data show that smoking, air pollution, chronic inflammation of the lungs, and long-term exposure to asbestos, dust, nickel, chromium and other metal substances may promote the occurrence of lung cancer. At the same time, lung cancer is also related to genetic factors, the body's autoimmune status, and metabolism.
  • lung cancer is currently divided into Small cell lung cancer (SCLC) and Non small cell lung cancer (NSCLC).
  • SCLC Small cell lung cancer
  • NSCLC Non small cell lung cancer
  • Non-small cell lung cancer accounts for more than 85% of lung cancer, and more than 85% of non-small cell lung cancer are advanced lung cancer. Therefore, the diagnosis of lung cancer, especially the diagnosis of non-small cell lung cancer has very important clinical significance.
  • non-small cell lung cancer includes: X-ray examination, bronchoscopy, cancer anti-blood index screening, lavage fluid/sputum cytology examination, and low-dose spiral CT (LDCT) screening that has been widely used in recent years .
  • these screening or diagnostic methods often have many problems, including low specificity and sensitivity, invasive screening or diagnosis, certain radiation damage, and low diagnostic compliance. Therefore, it is necessary to develop an efficient, accurate, and non-invasive method for large-scale non-small cell lung cancer screening or diagnosis for large-scale high-risk groups to help reduce the incidence of non-small cell lung cancer and improve the early screening of non-small cell lung cancer. Proportion.
  • tumor-specific molecular markers such as carcinoembryonic antigen (CEA), squamous cell carcinoma-associated antigen (SCC-Ag) and circulating microRNA (microRNA) ) and other detection plus auxiliary imaging examination, can improve the sensitivity of diagnosis.
  • CCA carcinoembryonic antigen
  • SCC-Ag squamous cell carcinoma-associated antigen
  • microRNA circulating microRNA
  • these molecular markers can be directly detected in blood and other body fluids. They have the advantages of simplicity, quickness, non-invasiveness, cheapness, and reproducible detection.
  • studies have reported some molecular markers that can be used for the diagnosis of lung cancer, in general, these markers have poor specificity and low reliability. Therefore, it is still necessary to explore efficient and specific new lung cancer molecular markers for early diagnosis of non-small cell lung cancer.
  • One object of the present invention is to provide a reagent for detecting piR-hsa-120522 in plasma used in the preparation of a screening or diagnostic kit for lung cancer and/or its precancerous lesions.
  • the invention provides a use of a reagent for detecting piR-hsa-120522 in plasma in the preparation of a screening or diagnostic kit for lung cancer and/or its precancerous lesions.
  • the lung cancer is non-small cell lung cancer.
  • the reagents are PCR reagents, nucleic acid constant temperature amplification reaction reagents, nucleic acid imprinting detection reagents, colloidal gold detection reagents, nucleic acid sequencing reagents or gene chip detection reagents.
  • the PCR reagent is a fluorescent quantitative PCR reagent.
  • the PCR reagent includes primers for PCR amplification of piR-hsa-120522, and the primer sequences are shown in SEQ ID NO.5 and SEQ ID NO.6.
  • the PCR reagent also includes a primer for reverse transcription of piR-hsa-120522, and the primer sequence is shown in SEQ ID NO.4.
  • the present invention also provides a screening or diagnostic kit for lung cancer and/or its precancerous lesions, which includes a reagent for detecting piR-hsa-120522 in plasma.
  • the reagents are PCR reagents, nucleic acid constant temperature amplification reaction reagents, nucleic acid imprinting detection reagents, colloidal gold detection reagents, nucleic acid sequencing reagents or gene chip detection reagents; preferably, the PCR reagents are fluorescent quantitative PCR reagents.
  • the PCR reagent includes primers for PCR amplification of piR-hsa-120522, and the primer sequences are shown in SEQ ID NO.5 and SEQ ID NO.6.
  • the PCR reagent also includes a primer for reverse transcription of piR-hsa-120522, and the primer sequence is shown in SEQ ID NO.4.
  • GGCGGCCCGGGTTCGACTCCCGGTGTGGGAAC SEQ ID NO. 1.
  • the present invention also provides a method for screening lung cancer and/or its precancerous lesions, the method comprising the following steps:
  • the lung cancer is non-small cell lung cancer.
  • the expression level of -hsa-120522 can achieve the purpose of screening or diagnosing patients with non-small cell lung cancer and precancerous lesions, and is beneficial to the early detection of non-small cell lung cancer.
  • the expression level of -120522 can achieve the purpose of screening or diagnosing patients with non-small cell lung cancer and its pre-lesions, and can effectively distinguish benign nodules from malignant tumor nodules.
  • the expression level of can achieve the purpose of screening or diagnosing patients with non-small cell lung cancer, which is of great significance for the effective screening or diagnosis of non-small cell lung cancer.
  • the screening or diagnostic kit provided by the present invention can quickly, efficiently and accurately screen or diagnose patients with lung cancer (especially non-small cell lung cancer), and provide a screening or diagnostic kit for lung cancer (especially non-small cell lung cancer). A new option.
  • Figure 1 is the ROC curve for detecting the expression level of piR-hsa-120522 in the plasma of patients with non-small cell lung cancer and precancerous lesions, benign nodules, and healthy people.
  • Figure 2 is the ROC curve for detecting the expression level of piR-hsa-120522 in the plasma of patients with non-small cell lung cancer and precancerous lesions and benign nodules.
  • Figure 3 is a ROC curve for detecting the expression level of piR-hsa-120522 in the plasma of patients with non-small cell lung cancer and benign nodules and healthy people.
  • the raw materials and equipment used in the present invention are known products obtained by purchasing commercially available products.
  • Example 1 The composition of non-small cell lung cancer screening or diagnosis kit
  • the non-small cell lung cancer screening or diagnosis kit provided by the invention consists of a fluorescent quantitative PCR reagent for detecting piR-hsa-120522 in human plasma.
  • the fluorescent quantitative PCR reagent includes primers for PCR amplification of piR-hsa-120522, and the primer sequences are shown in SEQ ID NO.5 and SEQ ID NO.6; it also includes reverse transcription of piR-hsa-120522 The primer, the primer sequence is shown in SEQ ID NO.4.
  • Example 2 Expression test of piR-hsa-120522 in plasma of patients with non-small cell lung cancer
  • a total of 73 patients with non-small cell lung cancer, 19 patients with precancerous lesions of non-small cell lung cancer, 49 healthy people, and 29 patients with benign pulmonary nodules were selected.
  • Plasma samples were taken from patients with non-small cell lung cancer, patients with precancerous lesions of non-small cell lung cancer, healthy people, and people with benign pulmonary nodules.
  • U6 was used as an internal reference gene to detect the target gene (see Table 1 for primer information).
  • GREEN I detects the expression of target genes in 168 plasma samples submitted for inspection. Specific steps are as follows:
  • scandrop100 ultra-trace nucleic acid protein analyzer
  • Use Aidlab s reverse transcription kit (TUREscript 1st Stand cDNA SYNTHESIS Kit) for reverse transcription, using a 20uL reaction system (different reaction systems are used according to the number of genes detected, the more genes, the larger the reaction system):
  • the reverse transcription reaction conditions are as follows:
  • the reverse transcription reaction conditions are as follows:
  • SPSS version 20.0 software was used for processing. The data were expressed as mean ⁇ standard deviation, the t test was used for comparison between two groups, and the one-way analysis of variance was used for comparison among multiple groups. P ⁇ 0.05 was regarded as a statistically significant difference.
  • Table 4 The plasma levels of patients with non-small cell lung cancer and precancerous lesions, benign nodules, and healthy people
  • the area under the ROC curve is 0.619, indicating that the method for screening or diagnosing patients with non-small cell lung cancer and precancerous lesions by detecting the expression level of piR-hsa-120522 in plasma has excellent Accuracy, Sensitivity and Specificity.
  • the area under the ROC curve is 0.705, indicating that the method for screening or diagnosing patients with non-small cell lung cancer and precancerous lesions by detecting the expression level of piR-hsa-120522 in plasma has excellent Accuracy, Sensitivity and Specificity.
  • the area under the ROC curve is 0.608, indicating that the method for screening or diagnosing patients with non-small cell lung cancer by detecting the expression level of piR-hsa-120522 in plasma has excellent accuracy and sensitivity. degree and specificity.
  • the present invention provides the use of a reagent for detecting piR-hsa-120522 in plasma in the preparation of a screening or diagnostic kit for non-small cell lung cancer.
  • the present invention found for the first time that compared with benign nodules and healthy people, the expression level of piR-hsa-120522 in the plasma of patients with non-small cell lung cancer and patients with precancerous lesions was significantly higher (P ⁇ 0.05), indicating that by detecting The expression level of piR-hsa-120522 can achieve the purpose of screening or diagnosing patients with non-small cell lung cancer, and is of great significance for the effective screening or diagnosis of non-small cell lung cancer.
  • the reagent for detecting piR-hsa-120522 in plasma can be used to prepare a non-small cell lung cancer screening or diagnosis kit, so as to realize effective screening or diagnosis of non-small cell lung cancer.
  • the screening or diagnostic kit provided by the present invention can quickly, efficiently and accurately screen or diagnose patients with lung cancer (especially non-small cell lung cancer), and provide a screening or diagnostic kit for lung cancer (especially non-small cell lung cancer). A new option.

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Abstract

本发明提供了检测血浆中piR-hsa-120522的试剂在制备非小细胞肺癌筛查或诊断试剂盒中的用途。本发明首次发现，与良性结节及健康人群相比，piR-hsa-120522在非小细胞肺癌和癌前病变患者血浆中的表达水平明显更高(P＜0.05)，说明通过检测血浆中piR-hsa-120522的表达水平能够达到筛查或诊断非小细胞肺癌患者的目的，对于实现非小细胞肺癌的有效筛查或诊断具有重要意义。

Description

检测血浆中piR-hsa-120522的试剂在制备肺癌筛查或诊断试剂盒中的用途 技术领域
本发明属于体外诊断试剂领域，具体涉及检测血浆中piR-hsa-120522的试剂在制备肺癌和/或其癌前病变筛查或诊断试剂盒中的用途。
背景技术
肺癌是最常见的恶性肿瘤之一，在全球范围内肺癌死亡率都属恶性肿瘤之首。肺癌是发病率和死亡率增长最快，对人群健康和生命威胁最大的恶性肿瘤之一。近50年来许多国家都报道肺癌的发病率和死亡率均明显增高，男性肺癌发病率和死亡率均占所有恶性肿瘤的第一位，女性发病率占第二位，死亡率占第二位。肺癌的发病机制至今尚未完全阐明，大量的研究资料表明吸烟、空气污染、肺部慢性炎症以及长期接触石棉、粉尘、镍、铬等金属物质都可能促进肺癌的发生。同时，肺癌也与遗传因素、机体自身免疫状态以及代谢相关。
从临床特征的角度看，目前国际上将肺癌分为小细胞肺癌Small cell lung cancer(SCLC)和非小细胞肺癌Non small cell lung cancer(NSCLC)。非小细胞肺癌占肺癌的85％以上，而非小细胞肺癌中85％以上都属中晚期肺癌。因此对肺癌的诊断，尤其是对非小细胞肺癌的诊断具有十分重要的临床意义。
临床上非小细胞肺癌的检查包含：X射线检查、支气管镜检查、癌抗血液指标筛查、灌洗液/痰液细胞学检查以及近几年大量使用的低剂量螺旋CT(LDCT)筛查。但这些筛查或诊断方法往往存在很多的问题，包含不高的特异性和灵敏度，有创侵入式的筛查或诊断，一定的辐射伤害以及不高的诊断依从性。因此，需要开发一种高效、准确、无创的方式对大规模的高危人群进行大范围的非小细胞肺癌筛查或诊断，帮助降低非小细胞肺癌的发病率，提高非小细胞肺癌早期筛出比例。
近年来，基于分子诊断的液体活检技术在肿瘤诊断中受到广泛关注，一些肿瘤特异的分子标志物如癌胚抗原(CEA)、鳞状细胞癌相关抗原(SCC-Ag)及循环微小RNA(microRNA)等的检测加上辅助的影像学检查，能提高诊断的灵敏度。这些分子标志物可在血液等体液中直接检测，具有简便、快捷、无创、廉价、可重复检测等优点，容易为病人接受，对肿瘤的临床诊断具有重要意义，有很好的临床应用前景。尽管研究已报道一些可用于肺癌诊断的分子标志物，但总体而言，这些标志物的特异性较差，可靠性较低。因此，仍需探索高效、特异的新肺癌分子标志物，用于非小细胞肺癌的早期诊断。
目前，还未见通过检测血浆中的piR-hsa-120522水平来筛查或诊断肺癌的报道。
发明内容
本发明的一个目的在于提供检测血浆中piR-hsa-120522的试剂在制备肺癌和/或其癌前病变筛查或诊断试剂盒中的用途。
本发明提供了一种检测血浆中piR-hsa-120522的试剂在制备肺癌和/或其癌前病变筛查或诊断试剂盒中的用途。
进一步地，所述肺癌为非小细胞肺癌。
进一步地，所述试剂为PCR试剂、核酸恒温扩增反应试剂、核酸印迹检测试剂、胶体金检测试剂、核酸测序试剂或基因芯片检测试剂。
进一步地，所述PCR试剂为荧光定量PCR试剂。
进一步地，所述PCR试剂中包括对piR-hsa-120522进行PCR扩增的引物，该引物序列如SEQ ID NO.5和SEQ ID NO.6所示。
进一步地，所述PCR试剂中还包括对piR-hsa-120522进行反转录的引物，该引物序列如SEQ ID NO.4所示。
本发明还提供了一种肺癌和/或其癌前病变筛查或诊断试剂盒，它包括用于检测血浆中piR-hsa-120522的试剂。
进一步地，所述试剂为PCR试剂、核酸恒温扩增反应试剂、核酸印迹检测试剂、胶体金检测试剂、核酸测序试剂或基因芯片检测试剂；优选的，所述PCR试剂为荧光定量PCR试剂。
进一步地，所述PCR试剂中包括对piR-hsa-120522进行PCR扩增的引物，该引物序列如SEQ ID NO.5和SEQ ID NO.6所示。
进一步地，所述PCR试剂中还包括对piR-hsa-120522进行反转录的引物，该引物序列如SEQ ID NO.4所示。
piR-hsa-120522，序列号：
GGCGGCCCGGGTTCGACTCCCGGTGTGGGAAC(SEQ ID NO.1)。
本发明还提供了一种筛查肺癌和/或其癌前病变的方法，所述方法包括以下步骤：
(1)利用上述的试剂盒检测受试者血浆中piR-hsa-120522；
(2)比较血浆中piR-hsa-120522：若受试者血浆中piR-hsa-120522表达水平显著高于良性结节和/或健康人群，则受试者为肺癌和/或其癌前病变患者。
进一步地，所述肺癌为非小细胞肺癌。
本发明中，“显著高于”表示差异具有统计学意义。
本发明首次发现，与良性结节及健康人群相比，piR-hsa-120522在非小细胞肺癌及癌前病变患者血浆中的表达水平明显更高(P＝0.032)，说明通过检测血浆中piR-hsa-120522的表达水平能够达到筛查或诊断非小细胞肺癌及癌前病变患者的目的，有利于非小细胞肺癌的早期发现。
本发明首次发现，与良性结节人群相比，piR-hsa-120522在非小细胞肺癌及癌前病变患者血浆中的表达水平明显更高(P＝0.017)，说明通过检测血浆中piR-hsa-120522的表达水平能够达到筛查或诊断非小细胞肺癌及前病变患者的目的，能够有效区分良性结节和恶性肿瘤结节。
本发明首次发现，与良性结节及健康人群相比，piR-hsa-120522在非小细胞肺癌患者血浆中的表达水平明显更高(P＝0.048)，说明通过检测血浆中piR-hsa-120522的表达水平能够达到筛查或诊断非小细胞肺癌患者的目的，对于实现非小细胞肺癌的有效筛查或诊断具有重要意义。
本发明提供的筛查或诊断试剂盒能够快速、高效、准确的对肺癌(特别是非小细胞肺癌)患者进行筛查或诊断，为肺癌(特别是非小细胞肺癌)筛查或诊断试剂盒提供了一种新的选择。
显然，根据本发明的上述内容，按照本领域的普通技术知识和惯用手段，在不脱离本发明上述基本技术思想前提下，还可以做出其它多种形式的修改、替换或变更。
以下通过实施例形式的具体实施方式，对本发明的上述内容再作进一步的详细说明。但不应将此理解为本发明上述主题的范围仅限于以下的实例。凡基于本发明上述内容所实现的技术均属于本发明的范围。
附图说明
图1为检测非小细胞肺癌及癌前病变患者与良性结节及健康人群血浆中piR-hsa-120522表达水平的ROC曲线。
图2为检测非小细胞肺癌及癌前病变患者与良性结节人群血浆中piR-hsa-120522表达水平的ROC曲线。
图3为检测非小细胞肺癌患者与良性结节及健康人群血浆中piR-hsa-120522表达水平的ROC曲线。
具体实施方式
本发明所用原料与设备均为已知产品，通过购买市售产品所得。
实施例1：非小细胞肺癌筛查或诊断试剂盒的组成
本发明提供的非小细胞肺癌筛查或诊断试剂盒由检测人血浆中的piR-hsa-120522的荧光定量PCR试剂组成。该荧光定量PCR试剂中包括对piR-hsa-120522进行PCR扩增的引物，该引物序列如SEQ ID NO.5和SEQ ID NO.6所示；还包括对piR-hsa-120522进行反转录的引物，该引物序列如SEQ ID NO.4所示。
实施例2：piR-hsa-120522在非小细胞肺癌患者血浆中的表达测试
1、临床资料
选取非小细胞肺癌患者73例，非小细胞肺癌癌前病变患者19例，健康人群49例，肺部良性结节人群29例。
2、仪器与试剂
主要仪器：analytikjena-qTOWER2.2型荧光定量PCR仪(德国)、SCILOGEX D3024R离心机(美国)、普通PCR仪analytikjena-Easycycler(德国)；超微量核酸蛋白测定仪scandrop100(德国)；移液器(美国bio-rad)。
主要试剂耗材：反转录试剂盒(TUREscript 1st Stand cDNA SYNTHESIS Kit)(艾德来)；
Figure PCTCN2022091993-appb-000001
预混液(中国DF)。10μL枪头(美国GCS)， 200μL枪头(美国GCS)，1ml枪头(美国GCS),200μL无RNA酶PCR反应管(AXGEN)；1.5Ml无RNA酶EP管(GCS)，低位白色PCR反应管(bio-rad)，光学封膜(bio-rad)；(枪头及EP管在使用前都进行灭菌处理并烘干备用)。
3、检测方法
分别取非小细胞肺癌患者、非小细胞肺癌癌前病变患者、健康人群、肺部良性结节人群的血浆样品，以U6为内参基因，检测目的基因(引物信息详见表1)，采用SYBR GREEN I检测送检168个血浆样品中目的基因的表达量。具体步骤如下：
(1)样本中总RNA的提取
(2)RNA检测结果
本实验用超微量核酸蛋白测定仪(scandrop100)检测RNA OD值，使用的是A260/A280比值。
(3)cDNA第一条链的合成
采用Aidlab公司反转录试剂盒(TUREscript 1st Stand cDNA SYNTHESIS Kit)进行反转录操作，采用20uL反应体系(根据检测基因多少而使用不同的反应体系，基因越多反应体系就越大)：
目的基因反转录(采用10uL反应体系)：
Figure PCTCN2022091993-appb-000002
反转录反应条件如下：
25℃       10min
42℃       50min
65℃       15min
反应结束后，得到cDNA，-80℃保存。
内参基因反转录(采取10ul体系)：
Figure PCTCN2022091993-appb-000003
反转录反应条件如下：
42℃           45min
65℃           15min
反应结束后，得到cDNA，-80℃保存。
(4)引物合成信息
表1.引物信息
Figure PCTCN2022091993-appb-000004
Figure PCTCN2022091993-appb-000005
(5)实时定量PCR(QPCR)反应
按照表2所示成分配置QPCR反应液，在PCR板离心机上4℃6000rpm离心30s。再置于定量PCR仪按照以下程序进行扩增。
荧光定量PCR程序：
步骤1、95℃-3分钟，
步骤2、95℃-10分钟，
步骤3、60℃-30s+读板，
转到步骤2，进行39个循环，
熔体曲线分析(60℃～95℃，+1℃/循环，保持时间4s)。
表2.QPCR反应液成分
Figure PCTCN2022091993-appb-000006
(6)目的基因相对表达量的计算
各个样品中目的基因相对表达量的计算公式如下：
Figure PCTCN2022091993-appb-000007
采用SPSS 20.0版软件进行处理。数据采用均数±标准差表示，两组较采用t检验，多组间比较采用单因素方差分析。以P<0.05为差异具有统计学意义。
4、实验结果
以U6为内参，利用2^-△△CT法计算各组人群血浆样品中的目标基因的表达量，结果如表3所示。
表3不同人群血浆中piR-hsa-120522表达水平测定结果
Figure PCTCN2022091993-appb-000008
Figure PCTCN2022091993-appb-000009
进一步分析非小细胞肺癌及癌前病变患者与良性结节及健康人群之间血浆样品中的目标基因的表达量，结果如表4所示。
表4非小细胞肺癌及癌前病变患者与良性结节及健康人群血浆中
piR-hsa-120522表达水平测定结果
Figure PCTCN2022091993-appb-000010
根据表4可以看出，与良性结节及健康人群相比，piR-hsa-120522在非小细胞肺癌及癌前病变患者血浆中的表达水平明显更高(P＝0.032)，说明通过检测血浆中piR-hsa-120522的表达水平能够达到筛查或诊断非小细胞肺癌及癌前病变患者的目的，有利于非小细胞肺癌的早期发现。
根据ROC曲线(图1)可以看出，ROC曲线下面积为0.619，说明通过检测血浆中piR-hsa-120522的表达水平来筛查或诊断非小细胞肺癌及癌前病变患者的方法具有优良的准确度、敏感度和特异性。
进一步分析非小细胞肺癌及癌前病变患者与良性结节人群之间血浆样品中的目标基因的表达量，结果如表5所示。
表5非小细胞肺癌及癌前病变患者与良性结节人群血浆中piR-hsa-120522表达水平测定结果
Figure PCTCN2022091993-appb-000011
根据表5可以看出，与良性结节人群相比，piR-hsa-120522在非小细胞肺癌及癌前病变患者血浆中的表达水平明显更高(P＝0.017)，说明通过检测血浆中piR-hsa-120522的表达水平能够达到筛查或诊断非小细胞肺癌及前病变患者的目的，能够有效区分良性结节和恶性肿瘤结节。
根据ROC曲线(图2)可以看出，ROC曲线下面积为0.705，说明通过检测血浆中piR-hsa-120522的表达水平来筛查或诊断非小细胞肺癌及癌前病变患者的方法具有优良的准确度、敏感度和特异性。
进一步分析非小细胞肺癌患者与良性结节及健康人群之间血浆样品中的目标基因的表达量，结果如表6所示。
表6非小细胞肺癌患者与良性结节及健康人群血浆中piR-hsa-120522表达水平测定结果
Figure PCTCN2022091993-appb-000012
根据表6可以看出，与良性结节及健康人群相比，piR-hsa-120522在非小细胞肺癌患者血浆中的表达水平明显更高(P＝0.048)，说明通过检测血浆中piR-hsa-120522的表达水平能够达到筛查或诊断非小细胞肺癌患者的目的，对于非小细胞肺癌的诊断具有重要意义。
根据ROC曲线(图3)可以看出，ROC曲线下面积为0.608，说明通过检测血浆中piR-hsa-120522的表达水平来筛查或诊断非小细胞肺癌患者的方法具有优良的准确度、敏感度和特异性。
综上，本发明提供了检测血浆中piR-hsa-120522的试剂在制备非小细胞肺癌筛查或诊断试剂盒中的用途。本发明首次发现，与良性结节及健康人群相比，piR-hsa-120522在非小细胞肺癌患者和癌前病变患者血浆中的表达水平明显更高(P<0.05)，说明通过检测血浆中piR-hsa-120522的表达水平能够达到筛查或诊断非小细胞肺癌患者的目的，对于实现非小细胞肺癌的有效筛查或诊断具有重要意义。检测血浆中的piR-hsa-120522的试剂能够用来制备非小细胞肺癌筛查或诊断试剂盒，实现非小细胞肺癌的有效筛查或诊断。本发明提供的筛查或诊断试剂盒能够快速、高效、准确的对肺癌(特别是非小细胞肺癌)患者进行筛查或诊断，为肺癌(特别是非小细胞肺癌)筛查或诊断试剂盒提供了一种新的选择。

Claims (12)

  1. 检测血浆中piR-hsa-120522的试剂在制备肺癌和/或其癌前病变筛查或诊断试剂盒中的用途。
  2. 根据权利要求1所述的用途，其特征在于：所述肺癌为非小细胞肺癌。
  3. 根据权利要求1或2所述的用途，其特征在于：所述试剂为PCR试剂、核酸恒温扩增反应试剂、核酸印迹检测试剂、胶体金检测试剂、核酸测序试剂或基因芯片检测试剂。
  4. 根据权利要求3所述的用途，其特征在于：所述PCR试剂为荧光定量PCR试剂。
  5. 根据权利要求3或4所述的用途，其特征在于：所述PCR试剂中包括对piR-hsa-120522进行PCR扩增的引物，该引物序列如SEQ ID NO.5和SEQ ID NO.6所示。
  6. 根据权利要求5所述的用途，其特征在于：所述PCR试剂中还包括对piR-hsa-120522进行反转录的引物，该引物序列如SEQ ID NO.4所示。
  7. 一种肺癌和/或其癌前病变筛查或诊断试剂盒，其特征在于：它包括用于检测血浆中piR-hsa-120522的试剂。
  8. 根据权利要求7所述的筛查或诊断试剂盒，其特征在于：所述试剂为PCR试剂、核酸恒温扩增反应试剂、核酸印迹检测试剂、胶体金检测试剂、核酸测序试剂或基因芯片检测试剂；优选的，所述PCR试剂为荧光定量PCR试剂。
  9. 根据权利要求8所述的筛查或诊断试剂盒，其特征在于：所述PCR试剂中包括对piR-hsa-120522进行PCR扩增的引物，该引物序列如SEQ ID NO.5和SEQ ID NO.6所示。
  10. 根据权利要求9所述的筛查或诊断试剂盒，其特征在于：所述PCR试剂中还包括对piR-hsa-120522进行反转录的引物，该引物序列如SEQ ID NO.4所示。
  11. 一种筛查肺癌和/或其癌前病变的方法，其特征在于：所述方法包括以下步骤：
    (1)利用权利要求7-10任一项所述的试剂盒检测受试者血浆中piR-hsa-120522；
    (2)比较血浆中piR-hsa-120522：若受试者血浆中piR-hsa-120522表达水平显著高于良性结节和/或健康人群，则受试者为肺癌和/或其癌前病变患者。
  12. 根据权利要求11所述的方法，其特征在于：所述肺癌为非小细胞肺癌。
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