Non-invasive marker and kit for diagnosis of lung squamous cell carcinoma patients among non-smoking or mild-smoking people

A non-invasive, non-smoking technology, applied to the determination/inspection of microorganisms, biochemical equipment and methods, DNA/RNA fragments, etc., which can solve the problems of unable to cover the population

Active Publication Date: 2017-05-10
MIRXES HANGZHOU BIOTECHNOLOGY CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Finally, the applicant grouped the population samples according to whether they smoked or not and the smoking index, and found two groups of microRNAs markers for the diagnosis of lung squamous cell carcinoma, but the smoking index of clinical samples needs to be limited, and these two groups of microRNAs markers cannot cover all people

Method used

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  • Non-invasive marker and kit for diagnosis of lung squamous cell carcinoma patients among non-smoking or mild-smoking people
  • Non-invasive marker and kit for diagnosis of lung squamous cell carcinoma patients among non-smoking or mild-smoking people
  • Non-invasive marker and kit for diagnosis of lung squamous cell carcinoma patients among non-smoking or mild-smoking people

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Example 1 Discovery and quantitative confirmation of plasma differential microRNA based on microRNA chip

[0030] 1. Experimental materials

[0031] 1. Instrument reagents

[0032] High-speed centrifuges, high-speed refrigerated centrifuges, and UV spectrophotometers were purchased from Eppendorf, Germany; NanoDrop 1000 spectrophotometers were purchased from Thermo Scientific, USA; Agilent2100Bioanalyzer System was purchased from Agilent, USA; ordinary PCR reaction instruments and real-time Fluorescence PCR instrument was purchased from American PE Company; DYCP-31B electrophoresis instrument was purchased from Beijing Liuyi Biotechnology Co., Ltd.; BIO-RAD gel imaging analysis was purchased from American Bio-Rad; ultra-low temperature refrigerator was purchased from Haier Group. mirvana TM paris TM The kit was purchased from Ambion Company of the United States; Agilent RNA 6000Pico Kit was purchased from Agilent Company of the United States; Trizol Total RNA Extra...

Embodiment 2

[0059] Example 2: ROC curve evaluation of diagnostic efficiency of plasma differential microRNA

[0060] The principle of ROC curve evaluation method:

[0061]The basic evaluation indicators of diagnostic tests include sensitivity, specificity, etc., and the comprehensive evaluation indicators include Youden index, ROC, AUC, etc. For the evaluation of diagnostic tests, one should first know the true category of the subjects, that is, which ones belong to the healthy group and which ones belong to the disease group. The standard for dividing the healthy group and the disease group is the gold standard (such as the histopathological diagnosis method recognized in this application). For the disease group and healthy group determined by the gold standard, the results of diagnostic tests can be divided into the following situations:

[0062] Positive (True Positive, TP); the diagnostic test is positive (consistent with the gold standard result);

[0063] Negative (True Negative,...

Embodiment 3

[0084] Example 3: Further verifying the accuracy of combined diagnosis of plasma differential microRNAs in the verification set

[0085] 1. Experimental materials

[0086] With embodiment 1.

[0087] 2. Experimental methods and results

[0088] 1. The extraction of plasma total RNA and the method of real-time fluorescent quantitative PCR are the same as in Example 1.

[0089] 2. In the verification set, based on the above binary logistic regression equation, the 4 plasma differential microRNAs (miR-432-3p, miR-371a-3p, miR-588, miR-676-5p) in the samples of the heavy smoking population in the verification set The standardized value of the content was transformed by binary logistic regression, and the logistic regression value of the four plasma differential microRNAs contents in the samples of heavy smoking population was calculated. Those who are lower than the optimal cut-off value of 0.496 are predicted to be healthy controls, and those who are higher than the optimal cu...

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Abstract

The invention discloses a non-invasive marker and a kit for diagnosis of lung squamous cell carcinoma patients among non-smoking or mild-smoking people, wherein the mild-smoking people refers to people with smoking index being lower than 200. The non-invasive marker is composed of the following microRNAs: miR-644a, miR-619-5p, miR-222-3p and miR-575, wherein the nucleotide sequence of the miR-644a is represented as the SEQ ID No.5; the nucleotide sequence of the miR-619-5p is represented as the SEQ ID No.6; the nucleotide sequence of the miR-222-3p is represented as the SEQ ID No.7; and the nucleotide sequence of the miR-575 is represented as the SEQ ID No.8. The microRNAs non-invasive marker is high in diagnosis accuracy and has high sensitivity and strong specificity.

Description

technical field [0001] The invention belongs to the field of molecular diagnosis, relates to non-invasive screening and diagnosis of lung squamous cell carcinoma, in particular to a non-invasive marker and a kit for diagnosing lung squamous cell carcinoma patients among non-smoking or light smokers. Background technique [0002] Lung cancer is the most common malignant tumor in the world, and its mortality rate ranks first among malignant tumors. The incidence of lung cancer in my country has been increasing year by year, with an average annual increase of nearly 2%. Lung cancer is divided into multiple tissue types according to different pathological characteristics, and the treatment measures are also different for different types of lung cancer tissue. According to the 2004 edition of the WHO classification, common histopathological types of lung cancer are divided into non-small cell carcinoma (NSCLC) and small cell carcinoma (SCLC). Non-small cell carcinomas are subdi...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/113C12N15/11
CPCC12Q1/6886C12Q2600/158C12Q2600/178
Inventor 郭守河郭帅其他发明人请求不公开姓名
Owner MIRXES HANGZHOU BIOTECHNOLOGY CO LTD
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