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Real-time Fluorescent Quantitative PCR for Detecting CA6 Virus

A real-time fluorescence quantitative and viral technology, applied in the field of detection, to achieve high specificity

Inactive Publication Date: 2017-02-08
SHANGHAI PUBLIC HEALTH CLINICAL CENT
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

So far, there have been several outbreaks of HFMD caused by CA6 around the world, but there are few reports on rapid and sensitive methods for specific detection of CA6, especially no specific real-time PCR (real-time PCR) detection Report on the method of CA6 virus

Method used

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  • Real-time Fluorescent Quantitative PCR for Detecting CA6 Virus
  • Real-time Fluorescent Quantitative PCR for Detecting CA6 Virus
  • Real-time Fluorescent Quantitative PCR for Detecting CA6 Virus

Examples

Experimental program
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Effect test

Embodiment 1

[0036] 1. The method for detecting CA6 virus by real-time fluorescence quantitative PCR, the steps are:

[0037] (1) Clinically collected specimens

[0038] From January to December 2012, a total of 605 patients with HFMD were collected from inpatients and outpatients of four hospitals: Shanghai Public Health Clinical Center, Xinhua Hospital Affiliated to Shanghai Jiaotong University School of Medicine, Shanghai Dongfang Hospital and Shanghai Changning District Central Hospital A total of 682 clinical specimens were collected, including 601 throat swabs and 81 stools; the hand, foot, and mouth diagnosis was based on the "Guidelines for Diagnosis and Treatment of Hand, Foot, and Mouth Disease 2010 Edition" issued by the Ministry of Health of China;

[0039] (2) Design primers

[0040] According to the capsid protein (VP1) coding sequence of CA6 included in the Genbank library and combined with the popular VP1 sequence of CA6 in Shanghai, primers and probes for CA6-specific rea...

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Abstract

The invention belongs to the field of nucleic detection and relates to a method for detecting CA6 virus with real-time fluorescent quantitative PCR. According to the method, a specific primer and a Taqman probe are designed on the basis of a CA6VP1 sequence and are used for detecting the CA6 virus rapidly, specifically and high-efficiently. By means of the method, gene segment of CA6 can be stably and effectively amplified and a highest detection sensitivity can reach 10 copius / [mu]l. In addition, the method is high in specificity and no false positive result is detected. The method for detecting the CA6 is quick, sensitive and reliable, can be further used for laboratory detection of a large-scale sample and an epidemiological inspection operation of the CA6 and is helpful to early-stage quick diagnosis in clinical practice.

Description

technical field [0001] The invention belongs to the detection field and relates to a nucleic acid detection method, in particular to a method for real-time fluorescent quantitative PCR detection of CA6 virus. Background technique [0002] The prior art discloses that hand, foot and mouth disease (HFMD) is a common infectious disease caused by human enterovirus (HEV); statistics show that the disease can occur all year round, 5 From July to July is the high-incidence period, and it is more common in children under 5 years old. Most of them have mild clinical symptoms, mainly fever and rash or herpes on the hands, feet, mouth and other parts. A small number of patients may have aseptic meningitis, encephalitis, Inflammation, acute flaccid paralysis (AFP), neurogenic pulmonary edema, and myocarditis, etc., the disease progresses rapidly in some severely ill children and can lead to death. Since 1997, there have been several outbreaks of HFMD in Malaysia, Singapore, Australia, ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/70C12Q1/68
CPCC12Q1/6851C12Q1/70C12Q2531/113C12Q2561/101C12Q2561/113
Inventor 胡芸文管文彩张晓玲宋志刚
Owner SHANGHAI PUBLIC HEALTH CLINICAL CENT
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