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Kit for detecting coronavirus RNA and method for detecting coronavirus RNA

A coronavirus and kit technology, applied in the field of molecular biology, can solve the problems of low specificity, false negatives, and inability to use rapid screening or detection, achieving high sensitivity, strong specificity, and satisfying clinical needs. The effect of rapid diagnosis and screening

Pending Publication Date: 2020-05-01
WUXI SHENRUI BIO PHARMA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the detection of new coronaviruses such as 2019-nCoV by qPCR requires a strict PCR amplification laboratory, and the qPCR method has certain false negative defects
In addition, although gene sequencing methods are more reliable, they cannot be used for rapid screening or detection
The antibody detection method needs to be detected after the patient has produced a sufficient immune response, and has the defect of low specificity of the detection result

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027]This embodiment discloses a kit for detecting coronavirus RNA. This kit is used for efficient, rapid and specific detection of SARS-CoV-1 and SARS-CoV-2 coronavirus RNA to quickly and effectively distinguish positive samples from negative samples.

[0028]Specifically, the kit includes: nuclease Cas13a, Cas13a guide RNA for specific detection of SARS-CoV-1 gene sequence and SARS-CoV-2 gene sequence, RNA fluorescent probe, reverse transcriptase, DNA polymerase , T7 RNA polymerase, and buffer. Among them, the nuclease Cas13a is selected from bacteria, yeast, insects, mammalian cell lines or other biological protein expression systems. More specifically, the nuclease Cas13a is selected from organisms of the following genera: Ciliates, Listeria, Corynebacterium, Sartorella, Legionella, Treponema, Nematogen, Eubacteria Genus, Streptococcus, Lactobacillus, Mycoplasma, Bacteroides, Flavivola, Flavobacterium, Sphaerochaeta, Azospirillum, Gluconobacter, Neisseria, Roche, Parvibaculum, Sta...

Embodiment 2

[0034]Based on the kit disclosed in Example 1, this example discloses a method for highly specific detection of the coronavirus RNA of targeted genes, namely SARS-CoV-1 and SARS-CoV-2, based on the kit. This method is used for specific detection of SARS-CoV-1 and SARS-CoV-2.

[0035]The method for detecting coronavirus RNA includes the following steps:

[0036]The RNA of the sample to be tested is extracted, and reverse transcriptase is added for in vitro reverse transcription to synthesize complementary strand cDNA. The sample to be tested is selected from swabs of alveolar lavage fluid, saliva, blood, plasma, serum, feces, urine, sputum, mucus, lymph, synovial fluid, spinal fluid, cerebrospinal fluid, skin or mucous membrane, or Any or several combinations.

[0037]T7 RNA polymerase is added for re-transcription. Bacteriophage T7 RNA polymerase is a DNA-dependent RNA polymerase, which has strict specificity for its respective double-stranded promoters. This enzyme catalyzes 5'→3' synthesis...

Embodiment 3

[0042]Based on the kit and method disclosed in Example 1, this example discloses an application based on the aforementioned kit and method. This method is based on a kit and is used for specific detection of SARS-CoV-1 and SARS-CoV-2.

[0043]For the technical solutions of the same part of the kit and method disclosed in this embodiment as those in the first or second embodiment, please refer to the above, and will not be repeated here.

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PUM

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Abstract

The invention provides a kit for detecting coronavirus RNA and a method for detecting the coronavirus RNA. The kit for detecting the coronavirus RNA comprises the following components: nuclease Cas13a, Cas13a guide RNA used for specifically detecting SARS-CoV-1 gene sequence and SARS-CoV-2 gene sequence, a RNA fluorescence probe, reverse transcriptase, DNA polymerase, T7RNA polymerase, and a buffer. The kit for detecting the coronavirus RNA and the method for detecting the coronavirus RNA disclosed by the invention are capable of realizing specific detection on coronaviruses with the SARS-CoV-1 gene sequence and the SARS-CoV-2 gene sequence, and have the advantages of being easy to operate, high in sensitivity, strong in specificity, free of PCR amplification conditions, good in stability,economical, practical and the like; and thus, needs of rapid clinical diagnosis and screening can be met.

Description

Technical field[0001]The present invention relates to the technical field of molecular biology, in particular to a kit and method for detecting coronavirus RNA.Background technique[0002]The 2019 Novel Coronavirus (2019-nCoV) was discovered due to a 2019 viral pneumonia case and was officially named SARS-CoV-2 by the World Health Organization on January 12, 2020. Coronavirus is a large virus family, known to cause colds and more serious diseases such as Middle East Respiratory Syndrome (MERS) and Severe Acute Respiratory Syndrome (SARS). It is very contagious and covers a wide range. Common signs of people infected with coronavirus include respiratory symptoms, fever, cough, shortness of breath, and difficulty breathing. In more severe cases, the infection can lead to pneumonia and severe acute respiratory syndrome. , Kidney failure, and even death. People of all ages can be infected, and the elderly and the weak and sick are more likely to be infected. Both the new coronavirus and t...

Claims

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Application Information

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IPC IPC(8): C12Q1/70C12Q1/6844C12R1/93
CPCC12Q1/6844C12Q1/701C12Q2521/107C12Q2563/107C12Q2521/327
Inventor 朱桂春盛青松
Owner WUXI SHENRUI BIO PHARMA