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Quadruple fluorescence quantitative detection kit for simultaneously detecting four human coronaviruses

A fluorescent quantitative detection and human coronavirus technology, applied in recombinant DNA technology, microorganism-based methods, and microbial determination/inspection, etc., can solve the problems of poor detection timeliness, high detection cost, and low detection efficiency. Realize the effects of multiple rapid detection, high detection sensitivity and convenient application

Active Publication Date: 2019-08-23
中华人民共和国无锡海关 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The purpose of the present invention is to solve the defects such as long time-consuming, low detection efficiency, high detection cost, and weak detection timeliness in the hCoVs detection process, and provide a quadruple fluorescence quantitative detection reagent for simultaneously detecting four kinds of human coronaviruses Box; it can simultaneously detect HCoV-229E, HCoV-NL63, HCoV-OC43, HCoV-HKUI four human coronaviruses in a single tube

Method used

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  • Quadruple fluorescence quantitative detection kit for simultaneously detecting four human coronaviruses
  • Quadruple fluorescence quantitative detection kit for simultaneously detecting four human coronaviruses
  • Quadruple fluorescence quantitative detection kit for simultaneously detecting four human coronaviruses

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0064] The sample to be tested is: throat swab.

[0065] Interpret the results corresponding to Table 3, the specific amplification curve is as follows figure 1 shown.

[0066] Such as figure 1 , the negative quality control or the blank control has no amplification curve, and the positive quality control has S-type amplification curves in the four detection channels of FAM, VIC, ROX and Cy5, and the samples to be tested only show S-type amplification curves in the FAM fluorescence channel. increase curve, and the Ct (or Cq) value ≤ 38, the sample to be tested is judged to be HCoV-229E positive.

Embodiment 2

[0068] The sample to be tested is: throat swab.

[0069] Interpret the results corresponding to Table 3, the specific amplification curve is as follows figure 2 shown.

[0070] Such as figure 2 , the negative quality control or the blank control has no amplification curve, and the positive quality control has S-type amplification curves in the four detection channels of FAM, VIC, ROX and Cy5, and the samples to be tested only show S-type amplification curves in the VIC fluorescence channel. increase curve, and the Ct (or Cq) value ≤ 38, the sample to be tested is judged to be HCoV-NL63 positive.

Embodiment 3

[0072] The sample to be tested is: throat swab.

[0073] Interpret the results corresponding to Table 3, the specific amplification curve is as follows image 3 shown.

[0074] Such as image 3 , the negative quality control product or the blank control has no amplification curve, and the positive quality control product has S-type amplification curves in the four detection channels of FAM, VIC, ROX and Cy5, and the samples to be tested only show S-type amplification curves in the ROX fluorescence channel. Increment curve, and Ct (or Cq) value ≤ 38, the sample to be tested is judged to be HCoV-OC43 positive.

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Abstract

The invention relates to a quadruple fluorescence quantitative detection kit for simultaneously detecting four human coronaviruses, and belongs to the technical field of human coronavirus detection. The human coronaviruses specifically include HCoV-229E / HCoV-NL63 / HCoV-OC43 / HCoV-HKUI. The detection kit specifically includes four specific primer pair and probe groups, a positive quality control product and a negative quality control product. Each specific primer pair and probe group comprises a specific primer pair and a probe sequence. The positive quality control product is specifically an artificially synthesized target sequence; the negative quality control product is deionized water or sterile water. According to the kit, the four human coronaviruses can be simultaneously detected by asingle tube, so that the detection specificity is high, the detection sensitivity is high, the detection time is short, and the defects of long consumed time, high detection cost, easy cross contamination and the like caused by one-by-one detection are avoided. At present, no similar related products are reported in the market. The kit has wide application prospects in the aspects of disease detection, infectious disease prevention and control, clinical treatment guidance and the like.

Description

technical field [0001] The invention relates to a quadruple fluorescence quantitative detection kit for simultaneously detecting four kinds of human coronaviruses, belonging to the technical field of detection of human coronaviruses. Background technique [0002] Human coronavirus (Human Coronaviruses, hCoVs) infection is one of the important pathogens that cause human respiratory diseases. The detection rate of coronaviruses in common respiratory pathogens is about 3%-10%. In adults, it is an important pathogen that causes clinical symptoms such as cough, fever, laryngitis, bronchitis and pneumonia. At present, there are six known human coronaviruses, human coronavirus 229E (Humancoronavirus 229E, HCoV-229E) and HCoV-OC43 discovered in the 1860s, and severe acute respiratory syndrome that appeared in Guangdong, China in 2003 (Severe acute respiratory syndrome, SARS) coronavirus SARS-CoV, novel human coronavirus NL63 (HCoV-NL63) isolated and identified in the Netherlands in...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/70C12Q1/686C12N15/11C12R1/93
CPCC12Q1/701C12Q1/686C12Q2600/16C12Q2563/107C12Q2545/113C12Q2537/143Y02A50/30
Inventor 耿合员朱娜蔡丽萍孙悦徐聪灵杨志俊汪圣强吴海磊谭文杰
Owner 中华人民共和国无锡海关
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