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Quadruple fluorescent quantitative detection kit capable of simultaneously detecting four human coronaviruses including 2019-nCoV

A 2019-ncov, fluorescence quantitative detection technology, applied in the field of detection, can solve the problems of poor detection timeliness, low detection efficiency, and high detection cost, and achieve the effect of multiple rapid detection, convenient application, and high detection sensitivity

Pending Publication Date: 2020-05-05
中华人民共和国无锡海关 +2
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] In order to solve the defects of long time-consuming, low detection efficiency, high detection cost and poor detection timeliness in the detection process of hCoVs, the present invention provides a four-way detection method for simultaneously detecting four kinds of human coronaviruses including 2019-nCoV. Heavy fluorescence quantitative detection kit, its detection is fast and specific

Method used

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  • Quadruple fluorescent quantitative detection kit capable of simultaneously detecting four human coronaviruses including 2019-nCoV
  • Quadruple fluorescent quantitative detection kit capable of simultaneously detecting four human coronaviruses including 2019-nCoV
  • Quadruple fluorescent quantitative detection kit capable of simultaneously detecting four human coronaviruses including 2019-nCoV

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0062] The sample to be tested is: nasopharyngeal swab.

[0063] Interpret the results corresponding to Table 3, the specific amplification curve is as follows figure 1 shown.

[0064] 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, it is judged that the sample to be tested is positive for MERS-CoV.

Embodiment 2

[0066] The sample to be tested is: nasopharyngeal swab.

[0067] Interpret the results corresponding to Table 3, the specific amplification curve is as follows figure 2 shown.

[0068] 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. If the Ct (or Cq) value is less than or equal to 38, it is judged that the sample to be tested is positive for 2019-nCoV.

Embodiment 3

[0070] The sample to be tested is: nasopharyngeal swab.

[0071] Interpret the results corresponding to Table 3, the specific amplification curve is as follows image 3 shown.

[0072] 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 fluorescent quantitative detection kit capable of simultaneously detecting four human coronaviruses including 2019-nCoV, and belongs to the technical field of detection. The detection kit is applied to RT-PCR / qRT-PCR detection. The detection kit specifically comprises four specific primer pairs, four probes and a positive quality control product. Each of four human coronaviruses correspond to one pair of specific primers and one probe sequence; and the positive quality control product is specifically an artificially synthesized target sequence. Sterile water or ultrapure water is taken as a negative quality control product. The quadruple fluorescent quantitative detection kit can be used for rapidly and simultaneously screening four human coronaviruses2019-nCoV / MERS-CoV / HCoV-OC43 / HCoV-HKUI and has the advantages of high detection sensitivity, strong specificity, simplicity in operation, convenience in application and the like, and the multiple rapid detection can be realized.

Description

technical field [0001] The invention relates to a quadruple fluorescence quantitative detection kit for simultaneously detecting four human coronaviruses including 2019-nCoV, in particular to four human A test kit for coronavirus belongs to the technical field of detection. Background technique [0002] Human coronavirus (Human Coronaviruses, hCoVs) infection is one of the important pathogens causing 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 seven known human coronaviruses, namely human coronavirus 229E (Humancoronavirus 229E, HCoV-229E) and HCoV-OC43 discovered in the 1860s, and severe acute respiratory syndrome (Severe acute respiratory syndrome) that appeared in 2003. syndrome, SARS) coronavirus SARS-CoV, a novel human coronavirus NL63 (HCoV-NL63) isol...

Claims

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Application Information

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IPC IPC(8): C12Q1/70C12Q1/6851C12N15/11
CPCC12Q1/701C12Q1/6851C12Q2600/16C12Q2600/166C12Q2531/113C12Q2537/143C12Q2563/107C12Q2545/113Y02A50/30
Inventor 耿合员田锋王凌冰朱娜孙悦汪圣强谭文杰
Owner 中华人民共和国无锡海关
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