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Method for detecting and identifying human polyomavirus with high sensitivity

A polyomavirus and virus technology, applied in the field of high-sensitivity detection and identification of human polyomavirus, can solve the problems that affect the in-depth study of the relationship between human polyomavirus and human diseases, low detection throughput, and retention

Inactive Publication Date: 2013-09-04
INST OF PATHOGEN BIOLOGY CHINESE ACADEMY OF MEDICAL SCI +1
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Problems solved by technology

However, the current detection method of human polyomavirus DNA is still at the stage of single virus-specific PCR or fluorescent quantitative PCR detection, and there is no report on a method covering multiple human polyomavirus detection.
Due to the low detection throughput, the in-depth study of the relationship between human polyomavirus and human diseases has been seriously affected

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  • Method for detecting and identifying human polyomavirus with high sensitivity

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Embodiment Construction

[0040] The invention includes simultaneous detection of one or more of 12 known human polyomaviruses, each genotype adopts VP1 gene region for detection. The specific embodiments described here are only used to explain the present invention, not to limit the present invention.

[0041] In the method for highly sensitive detection and / or identification of human polyomaviruses provided by the embodiments of the present invention, 12 kinds of human polyomaviruses to be detected include: BKPyV, JCPyV, KIPyV, WUPyV, MCPyV, HPyV6, HPyV7, TSPyV, HPyV9, MWPyV, STLPyV and HPyV12. Including the following steps:

[0042] (1) According to the full sequence of the human polyomavirus to be tested, obtain the VP1 gene region, select the specific conserved sequence of each type, and design a pair of PCR primers with 10 bases (ACGTTGGATG) at the 5' end of the primers Any combination of tag sequences can make the total length more than 30 bases, which is used to distinguish primers from probe...

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Abstract

The invention provides a method for detecting and identifying human polyomavirus with the high sensitivity. According to the method, elimination of PCR (polymerase chain reaction) amplification product pollution is used as a precondition, and multiplex PCR-mass spectrometry is used as a platform to detect and / or identify the human polyomavirus with the high sensitivity. 12 kinds of human polyomaviruses detected by the method include BKPyV (BK polyomavirus), JCPyV (JC polyomavirus), KIPyV (Ki polyomavirus), WUPyV (WU polyomavirus), MCPyV (merkel cell polyomavirus), HPyV6 (human polyomavirus6), HPyV7 (human polyomavirus7), TSPyV (trichodysplasia spinulosa-associated polyomavirus), HPyV9 (human polyomavirus9), MWPyV (MW polyomavirus), STLPyV (STL polyomavirus) and HPyV12 (human polyomavirus12), and a detection part is a late coding region VP1 of coding virus capsid protein in a virus genome.

Description

technical field [0001] The invention relates to a method for detecting pathogenic microorganisms, in particular to a method for detecting and identifying human polyomaviruses with high sensitivity. Background technique [0002] Polyomavirus is a double-stranded DNA virus with a genome of about 4700-5400 base pairs, encoding structural proteins and antigens. Since Gross discovered in 1953 that murine polyomaviruses can cause tumors in mice, polyomaviruses have been suspected to be the causative factors of human tumors. However, although polyomavirus infection can induce tumors in animal models, no definite evidence of causing human tumors has been found. In animals, polyomavirus DNA integration into the host genome often precedes tumor formation. [0003] As of March 2013, 12 human polyomaviruses have been discovered. BK polyomavirus (BKPyV) and JC polyomavirus (JCPyV), reported in 1971, are regarded as the earliest human polyomaviruses discovered. BKPyV is thought to be ...

Claims

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Application Information

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IPC IPC(8): C12Q1/70C12Q1/68C12R1/93
Inventor 彭俊平郭军华金奇
Owner INST OF PATHOGEN BIOLOGY CHINESE ACADEMY OF MEDICAL SCI
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