Biomarker for saliva test of 2019-nCoV and application of biomarker

A 2019-ncov and 2019ncov technology, applied in the field of molecular biology, can solve problems such as difficult constant standardization, false negative detection, and operators' susceptibility to infection, and achieve simple sampling, high sensitivity and specificity, and good reliability and stability effects

Inactive Publication Date: 2020-07-17
上海力拜生物科技有限公司 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Especially at the current urgent moment when lung lavage fluid cannot be carried out on a large scale, almost all virus samples from patients are collected through nasopharyngeal swabs
The quality of nasopharyngeal swab sampling varies with the operation of the experimenter, and it is difficult to be standardized constantly, which is one of the important reasons for false negative detection
At present, the positive rate of new coronavirus nucleic acid detection is only about 30%-40%.
In addition, during nasopharyngeal swab sampling operations, operators are vulnerable to infection

Method used

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  • Biomarker for saliva test of 2019-nCoV and application of biomarker
  • Biomarker for saliva test of 2019-nCoV and application of biomarker
  • Biomarker for saliva test of 2019-nCoV and application of biomarker

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0093] The purification of trace nucleic acid in the saliva of embodiment 1

[0094] 1. Sample source:

[0095] Shanghai Public Health Clinical Center

[0096] Collection method: before collection, the patient rinses mouth with water 3-4 times to remove oral mucus and food residue. After gargling, stand still for 3-5 minutes, secrete 1.5-2 ml of saliva under the tongue or from both cheeks, and spit it into a saliva collection tube with a cover. The collected samples should be placed in a refrigerator at 4 degrees Celsius immediately.

[0097] A total of 25 cases were collected.

[0098] 2. Use the following steps to purify trace amounts of nucleic acid in saliva:

[0099] 1) Rinse mouth with water 3-4 times to remove food residue and mucus in the mouth; stand still for 3 minutes.

[0100] 2) Spit 1.5-2 ml of saliva into a capped experimental tube treated with high temperature and high pressure.

[0101] 3) Transfer the sample to a 1.5 ml high temperature and high pressure...

Embodiment 2

[0112] Embodiment 2 purification result analysis

[0113] The purified saliva supernatant nucleic acid generates cDNA (Qiagen reverse transcription kit) after reverse transcription, and then uses the specific amplification primer of coronavirus gene S protein to the amplification of the nucleic acid sample PCR that obtains in embodiment 1 (PCR), the sequence of the specific amplification primer with coronavirus gene S protein is:

[0114] Upstream primer: (SEQ ID NO: 1): Downstream primer: (SEQ ID NO: 2):

[0115] Table 1 Amplification reaction system

[0116] dNTP (2.5μM) 160μL 10X EX buffer 2.0 μL specific primer 200μL template DNA 1.0 μL Teq DNA polymerase 10μL PCR buffer (10 times working concentration) 200μL

[0117] (2) The results of PCR were carried out by agarose gel electrophoresis, and the results were as follows: figure 1 As shown, the result proves that:

[0118] According to the method of the present inventio...

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Abstract

The invention provides a biomarker for saliva test of 2019-nCoV and an application of the biomarker, and provides an application of a coronavirus S protein gene as the biomarker in preparation of a 2019-nCoV saliva test product or a 2019-nCoV coronavirus pneumonia auxiliary saliva diagnostic product. The inventors find for the first time the role of the 2019-nCoV gene as the biomarker in the preparation of the 2019-nCoV saliva test product. The product can quickly detect the 2019-nCoV infection, samples are easy to obtain, the detection efficiency is high, the speed is fast, the risk of infection of operators is reduced, and the product has very high sensitivity and specificity.

Description

technical field [0001] The invention relates to the technical field of molecular biology, in particular to a 2019-nCoV novel coronavirus saliva detection biomarker and its application. Background technique [0002] Due to the long incubation period of the coronavirus and the large differences in the performance of infected individuals, it has brought great challenges to rapid / accurate diagnosis. From the "Diagnosis and Treatment Plan for Novel Coronavirus Infected Pneumonia (Trial Version 6)" published by the National Health and Medical Commission, we can see the sum of multiple detection methods, which are: [0003] 1) Epidemiological history. It is mainly to determine whether there has been contact with a suspected person or place. [0004] 2) Personal physiological condition. It mainly includes whether there is fever or respiratory symptoms. [0005] 3) Blood biochemical testing. It is mainly tested by drawing blood, such as the total number of white blood cells or l...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/70C12Q1/686C12Q1/6806C12R1/93
CPCC12Q1/6806C12Q1/686C12Q1/701C12Q2600/118C12Q2521/107C12Q2565/125
Inventor 唐欣朱民雷芙蓉
Owner 上海力拜生物科技有限公司
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