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Extraction-free SARS-CoV-2 nucleic acid detection method and kit

A coronavirus and kit technology, applied in the fields of biotechnology and medicine, can solve problems such as time-consuming

Inactive Publication Date: 2020-05-19
杭州丹威生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The processing of a single sample often takes tens of minutes, and when a large number of samples need to be processed, it takes even longer than the PCR detection reaction

Method used

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  • Extraction-free SARS-CoV-2 nucleic acid detection method and kit
  • Extraction-free SARS-CoV-2 nucleic acid detection method and kit
  • Extraction-free SARS-CoV-2 nucleic acid detection method and kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] Example 1. Development of an extraction-free fluorescent PCR detection kit for novel coronavirus

[0036] 1. Design of primers and probes

[0037] Through the use of primer probe online design tools for the new coronavirus genome sequence published by the National Bioinformatics Center / National Genome Science Data Center http: / / www.yeastgenome.org / cgi-bin / web-primer Design specific primers and probes for ORF1ab gene and N gene. Considering specificity and amplification efficiency, the final selection of two sets of primer probe sequences is as follows:

[0038] The sequences of the ORF1ab gene-specific forward and reverse primers are 5'-GGCTTCACATATGTATTGTTC-3' and 5'-GCTCAAACTCTTCTTCTTCAC-3'; the sequence of the ORF1ab gene-specific probe is 5'-TCACCTTCTTCTTCATCCTCATCTGG-3', In addition, the two ends of the probe are respectively labeled with a fluorescence generating group FAM and a fluorescence quenching group BHQ1;

[0039] The sequences of the N gene-specific forward an...

Embodiment 2

[0051] Example 2. Detection of actual clinical samples

[0052] 1. Types of clinical samples The clinical samples used in this example come from throat swab samples collected by the Zhejiang Provincial Center for Disease Control and Prevention. The swabs collected are stored in normal saline or virus preservation solution. There are 301 cases, of which 105 are positive samples. 106 negative samples.

[0053] 2. The reference kit uses the kit used by the Zhejiang Provincial Center for Disease Control and Prevention as the reference reagent.

[0054] 3. Test results

[0055] 301 cases were selected for testing, of which 2 samples were thick sputum and the type of samples required by the kit were inconsistent and therefore were eliminated. 4 cases were excluded because the internal standard was not found and the test results were invalid. A total of 295 valid cases were excluded. All valid results are included in statistical analysis.

[0056]

[0057] 4. Result analysis

[0058] The resu...

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Abstract

The invention provides an extraction-free method and a kit for qualitatively detecting SARS-CoV-2 by fluorescence polymerase chain reaction (fluorescence PCR), and belongs to the field of biotechnology and medicine. The method integrates sample nucleic acid release and amplification. The kit comprises a lysis solution and a rapid enzyme, can directly lyse and amplify respiratory secretions of a patient, make a whole detection process fast and simple, reduce chance of mutual contamination, obtain accurate and reliable PCR test results, can diagnose presence of SARS-CoV-2 in patient secretions as soon as possible, and has important application value in the fields of disease monitoring and clinical diagnosis.

Description

Technical field: [0001] The present invention relates to the fields of biotechnology and medicine, in particular to a method and a kit for detecting a novel coronavirus. It is suitable for the detection and clinical auxiliary diagnosis of new coronavirus infection. Background technique: [0002] Novel coronavirus pneumonia is a disease caused by a new type of coronavirus (Severe acute respiratory syndromecoronavirus 2, SARS-CoV-2). In December 2019, patients with unexplained pneumonia appeared in some medical institutions in Wuhan. Wuhan continued to monitor influenza and related diseases. On January 7, 2020, a new type of coronavirus was detected in the laboratory and the entire genome of the virus was obtained. sequence. At present, the source of infection is mainly patients infected by the new type of coronavirus. Invisible infections may also become the source of infection. The spread is mainly through respiratory droplets and contact. The population is generally susceptibl...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/70C12Q1/686C12R1/93
CPCC12Q1/686C12Q1/701C12Q2600/166C12Q2521/107C12Q2545/101C12Q2561/101
Inventor 赵丹蕊郭兴中陈文方剑秋
Owner 杭州丹威生物科技有限公司
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