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Kit and method for detecting 25 RNA viruses in respiratory tract by NGS targeted probe capture method

An RNA virus and respiratory tract technology, applied in the field of kits for detecting 25 respiratory tract RNA viruses by NGS targeted probe capture method, can solve the problems of high cost and low false positive sensitivity, and achieve low cost, strong specificity and coverage. wide effect

Inactive Publication Date: 2020-08-25
南京实践医学检验有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, mNGS technology also has certain limitations in the detection of pathogenic microorganisms, such as high cost, false positives and low sensitivity.

Method used

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  • Kit and method for detecting 25 RNA viruses in respiratory tract by NGS targeted probe capture method
  • Kit and method for detecting 25 RNA viruses in respiratory tract by NGS targeted probe capture method
  • Kit and method for detecting 25 RNA viruses in respiratory tract by NGS targeted probe capture method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0049] Example 1 Preparation of 25 Respiratory Virus Detection Kits

[0050] The kit consists of a probe combination working solution, the sequence of which is shown in SEQ ID NO.1 to SEQ ID NO.50, and a probe preservation solution TE buffer.

Embodiment 2

[0051] Clinical detection and result analysis of embodiment 2 virus

[0052] Virus detection is performed in the following steps:

[0053] 1) Extraction of total RNA: Use QIAamp Viral RNA Mini Kit to extract RNA from nasopharyngeal swab or alveolar lavage fluid of the sample to be tested. In order to meet the library construction, the concentration of RNA required for extraction is not less than 5ng / ul And the purity is qualified.

[0054] 2) Removal of viral rRNA: using a ribosomal RNA removal kit. First add rRNA probes for hybridization and capture, then add RNase and DNase to digest rRNA and probes to obtain mRNA. The specific operation is as follows: Take an appropriate amount of 0.05-1 μg total RNA into a Nuclease-free microcentrifuge tube and dilute with Nuclease-free H2O to 11 μL (can be adjusted according to the actual situation), take out the rRNA Probe (H / M / R) and Probe Buffer from -20°C, thaw in an ice bath, vortex and mix well, add 1ul of rRNA Probe and 3ul of ...

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Abstract

The invention relates to the technical field of genetic engineering, and discloses a kit and a method for detecting 25 RNA viruses in the respiratory tract. The method is based on an NGS targeted probe capture technology, and a capture probe combination with strong specificity is designed for the 25 RNA viruses in the human respiratory tract. The kit and the technology for detecting the 25 RNA viruses in the respiratory tract have the advantages of targeted specificity, high sensitivity and wide coverage, and provide an important detection means for respiratory virus infection.

Description

technical field [0001] The invention relates to the technical field of genetic engineering, in particular to a kit and a method for detecting 25 kinds of RNA viruses in the respiratory tract by an NGS targeted probe capture method. Background technique [0002] Adults and children are disproportionately affected by sudden death. About 15-86% of children under the age of 5 died suddenly due to infection, of which 20-48% were caused by respiratory virus infection. Thus, viral infections of the respiratory tract are one of the leading causes of sudden death in children well before heart attack, and long before central nervous system involvement. These are the same viruses that cause upper or lower respiratory infections in the general population, such as influenza A and B (FluA or FluB), respiratory syncytial virus (RSV), parainfluenza, metapneumovirus, adenovirus (ADV) and Rhinoenterovirus, etc. The epidemiology associated with respiratory viruses is markedly seasonal with ...

Claims

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Application Information

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IPC IPC(8): C12Q1/70C12Q1/6813C12N15/11
CPCC12Q1/6813C12Q1/701C12Q2531/113Y02A50/30
Inventor 何贵伦张鹏唐春花邢宽何志健谢珍
Owner 南京实践医学检验有限公司