Method for simultaneously detecting twelve kinds of common respiratory viruses

A respiratory and virus technology, applied in the field of virus detection by molecular biology methods, can solve problems such as low diagnostic efficiency, high biosafety risk, and uncertain sensitivity, so as to save production costs and detection costs, improve detection efficiency, and reduce false positives. Positivity Effect

Inactive Publication Date: 2015-02-11
FUJIAN INT TRAVEL HEALTH CARE CENT +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The symptoms of infection caused by respiratory viruses are relatively similar, and it is difficult to make a definite diagnosis by clinical symptoms and conventional detection methods. Therefore, a highly sensitive and specific method for screening is urgently needed
[0003] At present, the commonly used detection methods for respiratory viruses in my country mainly include the following: 1. Electron microscope examination directly conducts morphological identification of viruses in samples. Disadvantages: ① The diagnostic efficiency is low, and some viruses are difficult to distinguish directly from the morphology; ② expensive equipment and experienced operators are required
2. Virus isolation and culture is the "gold standard" for virus diagnosis. Disadvantages: ① Time-consuming and labor-intensive, and require experienced operators; ② Biosafety risk is high
Disadvantages: ① prone to false positives; ② uncertain sensitivity; ③ false negative results when viral antigens mutate
However, there are also the following disadvantages: ①The throughput is low, generally only one virus can be detected at a time, and there is fluorescence interference affecting the detection results during multiple fluorescence detection; ②The detection cost is relatively high

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  • Method for simultaneously detecting twelve kinds of common respiratory viruses
  • Method for simultaneously detecting twelve kinds of common respiratory viruses
  • Method for simultaneously detecting twelve kinds of common respiratory viruses

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Embodiment Construction

[0058] In order to make the present invention easier to understand, the present invention will be further described below in conjunction with specific examples. It should be understood that these examples are only used to illustrate the present invention and are not intended to limit the scope of the present invention. The specific experimental methods not mentioned in the following examples are generally carried out according to conventional experimental methods.

[0059] Specific implementation one:

[0060] Sample nucleic acid extraction

[0061] Collect patient samples and extract total nucleic acid: collect nasopharyngeal swabs and isolated cultures of sputum from patients, and extract total nucleic acids from the isolated cultures. The invention designs specific nucleotide sequences of specific primers for specifically detecting 12 common respiratory viruses.

[0062] The reverse transcription primers for 12 kinds of respiratory viruses and 1 kind of internal control...

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Abstract

The invention discloses a method for simultaneously detecting twelve kinds of common respiratory viruses. According to the method, primers and probes are designed according to gene conservative areas of the twelve kinds of common respiratory viruses, namely influenza A virus, influenza B virus, influenza C virus, parainfluenza virus type 1, parainfluenza virus type 2, parainfluenza virus type 3, rhinovirus, Bocavirus, adenovirus, coronavirus, metapneumovirus and respiratory syncytial virus, nucleic acid fragments of samples to be measured are extracted for amplifying, and finally, the samples are separated by using a capillary electrophoresis method. The method disclosed by the invention has the advantages of low required sample size, high sensitivity and accuracy, good specificity and low cost; the defects that the conventional single tube multiplex fluorescence PCR (Polymerase Chain Reaction) detection primers are difficult to design, and multicolor fluorescence mutually intervenes and is not easy to part are overcome, the defects that a chip detection method is tedious in operation, high in detection cost and the like are also overcome, and a new method is provided for screening the respiratory viruses.

Description

technical field [0001] The invention belongs to the field of virus detection by molecular biological methods, and in particular relates to a method for detecting 12 kinds of respiratory viruses. Background technique [0002] Respiratory viruses are a large class of viruses that can invade the respiratory tract and mainly cause lesions in tissues and organs outside the respiratory tract. According to statistics, more than 90% of acute respiratory infections are caused by viruses, among which respiratory viruses are the most common, including influenza virus, rhinovirus, respiratory syncytial virus, parainfluenza virus, adenovirus, etc. Moreover, 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 infections have the characteristics of short incubation period, acute onset, strong infectivity, and rapid transmission, which...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/70C12Q1/68C12N15/11C12R1/93
CPCC12Q1/701C12Q2600/112C12Q2600/16C12Q2600/166
Inventor 王宇平王凌冰张建明黄野能高博何冰
Owner FUJIAN INT TRAVEL HEALTH CARE CENT
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