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Primer combination and method for detecting common respiratory viruses

A primer combination and detection method technology, applied in biochemical equipment and methods, recombinant DNA technology, and microbial assay/inspection, etc., can solve problems such as disappearance of fluorescent signal, inability to obtain virus sequence, similarity, etc.

Pending Publication Date: 2021-07-30
广东科蓝生物技术有限公司
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the symptoms of infection caused by respiratory viruses are often similar, and it is difficult to make a definite diagnosis based on clinical symptoms and routine detection methods.
[0004] At present, the methods commonly used to detect respiratory viruses are fluorescent quantitative PCR, etc. These methods cannot obtain the sequence of the virus. If the virus undergoes a mutation of a few bases, it may lead to missed detection such as the disappearance of the fluorescent signal.

Method used

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  • Primer combination and method for detecting common respiratory viruses
  • Primer combination and method for detecting common respiratory viruses
  • Primer combination and method for detecting common respiratory viruses

Examples

Experimental program
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Embodiment 1

[0049] The detection method of embodiment 1 common respiratory virus

[0050] Please refer to figure 1 , is a flow chart of the method for detecting common respiratory viruses in this embodiment, comprising the following steps:

[0051] 1. Sample pretreatment

[0052] In this embodiment, a total of 253 throat test sub-samples with known clinical results were selected for testing, 233 of which were samples of known infection types, and the other 20 were negative control samples (Table 1).

[0053] Table 1 The type and number of samples tested

[0054]

[0055] Take 300ul of sputum from each sample into a new 1.5ml centrifuge tube, add 300ul of sputum digestion liquid WT, vortex and mix, incubate at 37°C for 30min, vortex every 5-10min during digestion, and finally make the sputum sample homogeneous qualitative.

[0056] 2. RNA extraction

[0057] A common commercial RNA extraction kit (Tiangen or Qiagen) was used to extract nucleic acid from each sample.

[0058] (...

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Abstract

The invention discloses a primer combination and a method for detecting common respiratory viruses. The primer combination comprises two or more of 12 pairs of primers for detecting 15 common respiratory viruses. The detection method comprises the following steps: performing high-throughput sequencing on a captured sequence of a to-be-detected sample, and comparing the sequence with characteristic sequences of specific region nucleic acids of 15 common respiratory viruses by using a bioinformatics analysis method, so that whether the sequence of the to-be-detected sample is matched with the 15 common viruses or not can be very accurately judged. The primer combination has the advantages of high accuracy and high flux.

Description

technical field [0001] The invention belongs to the technical field of gene detection, and more specifically, the invention relates to a combination of primers and a method for detecting 15 common respiratory viruses. Background technique [0002] Respiratory viruses refer to viruses that mainly invade the respiratory tract, proliferate in the epithelial cells of the respiratory tract mucosa, and cause local infection of the respiratory tract or lesions of tissues and organs other than the respiratory tract, including common influenza viruses, human metapneumoviruses, syncytial viruses, parainfluenza viruses, and coronaviruses Wait. According to statistics, clinically about 90% to 95% of acute respiratory infections are caused by such respiratory viruses. [0003] In recent years, new pathogenic respiratory viruses have been continuously discovered, such as metapneumovirus, coronavirus, boca virus, etc., which pose a great threat to human health. Most acute respiratory inf...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/70C12Q1/6869C12Q1/686C12N15/11
CPCC12Q1/701C12Q1/686C12Q1/6869C12Q2600/16C12Q2535/122Y02A50/30
Inventor 韩凤章李雪多
Owner 广东科蓝生物技术有限公司
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