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Specific primers, probes and kits for detecting echovirus30 virus

A kit and specific technology, applied in the determination/testing of microorganisms, biochemical equipment and methods, DNA/RNA fragments, etc., can solve the problems of long detection cycle, high detection technology requirements, poor sensitivity, etc., and shorten the detection time cycle effect

Active Publication Date: 2018-03-27
SHANGHAI BIOGERM MEDICAL TECH CO LTD
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  • Abstract
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  • Claims
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AI Technical Summary

Problems solved by technology

[0009] Based on this, in order to overcome the above-mentioned defective of prior art, the present invention provides a kind of specific primer, probe and fluorescent quantitative PCR detection kit of detection Echovirus30 virus, solved prior art to detection technology requirement is high, and detection period Shortcomings of long, poor sensitivity

Method used

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  • Specific primers, probes and kits for detecting echovirus30 virus
  • Specific primers, probes and kits for detecting echovirus30 virus
  • Specific primers, probes and kits for detecting echovirus30 virus

Examples

Experimental program
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Effect test

Embodiment 1

[0025] A kind of real-time quantitative fluorescent PCR detection kit of embodiment 1 Echovirus30 virus

[0026] The preparation of the detection kit described in this embodiment comprises the following steps:

[0027] 1. Determine the detection target and specific primers and probes

[0028] Sequencing a total of 26 strains of ECHO30 viruses preserved in the Guangdong Provincial Germ and Virus Stock Bank, combined with a total of 29 ECHO30 virus sequences in GenBank, and determining the relatively conserved fragment SEQ ID NO: 1 by sequence comparison of the target gene VP1 as a detection the target sequence;

[0029] Using Bioedit and PrimerPremier5.0 and other software for bioinformatics analysis, design qPCR primer pairs SEQ ID NO:2 and SEQ ID NO:3 and Taqman probe SEQ ID NO:4 for the sequence of SEQ ID NO:1. Primers and Taqman probes were sent to Biological Company for synthesis.

[0030] The specific sequence is as follows:

[0031] Detected target sequence SEQ ID ...

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Abstract

The invention discloses a specific primer, a probe and a kit for detecting the Echovirus 30 virus, and belongs to the technical field of biological detection. The specific primer is composed of basic groups shown in the SEQ ID NO:2 and the SEQ ID NO:3. The probe is composed of basic group shown in the SEQ ID NO:4. According to the kit for detecting the Echovirus 30 virus, the specific primer, the probe and an optimized detection system are adopted, sensitivity and specificity can both reach 100%, and a sample with the content as low as 27.7copy / ul can be detected; a PCR product does not need to be sequenced, the detection cycle of an existing method is greatly shortened, and meanwhile detection sensitivity is improved. By means of the kit, a patient infected with the Echovirus 30 virus can be diagnosed at the early stage, and the blank of the kit for the Echovirus 30 virus in China is filled.

Description

technical field [0001] The invention belongs to the field of biotechnology, in particular, the invention relates to a specific detection primer, a probe and a fluorescent quantitative PCR detection kit for viral encephalitis-associated virus Echovirus30. Background technique [0002] Human Echovirus 30 (ECHO30) belongs to type B enteroviruses and is one of the main pathogens causing aseptic meningitis in children and adults. Among the cases of viral encephalitis in children, the cases caused by ECHO30 virus accounted for 80-93%. The virus is a non-enveloped virus that can survive stably under harsh environmental conditions such as high salt, high pH and 37°C. Human enteroviruses can still excrete viruses from the respiratory tract and intestines within a few weeks after being infected with human enteroviruses, which can cause virus transmission, infection and even outbreaks. [0003] In recent years, outbreaks or epidemics of viral encephalitis caused by ECHO30 have been r...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/70C12Q1/6851C12N15/11
CPCC12Q1/6851C12Q1/70C12Q2561/113C12Q2563/107
Inventor 陆靖柯昌文李晖郑焕英
Owner SHANGHAI BIOGERM MEDICAL TECH CO LTD
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