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Primer and probe sequence for human metapneumovirus nucleonic acid detection

A human metapneumovirus, primer sequence technology, applied in biochemical equipment and methods, microbial determination/inspection, DNA/RNA fragments, etc., can solve the problems of RT-PCR sensitivity, low sensitivity, etc.

Inactive Publication Date: 2011-04-27
中国疾病预防控制中心病毒病预防控制所
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Problems solved by technology

In addition, some people think that RT-PCR based on the N gene may be more sensitive
Therefore, Cotes et al. believe that RT-PCR based on the N gene and L gene is most suitable for the detection of hMPV, and Maertzdorf et al. prove that the primers based on the N gene may be less sensitive to B-type viruses due to mismatching

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  • Primer and probe sequence for human metapneumovirus nucleonic acid detection
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  • Primer and probe sequence for human metapneumovirus nucleonic acid detection

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Embodiment Construction

[0007] 1. Selection and optimization of primers and probes:

[0008] For the primer probe sequence of N gene, the sequence homologous to the 001 strain (its GenBank accession number is af371337) firstly reported by Dutch scholar Van den Hoogen was selected, and the probe used MGB probe technology to shorten the oligo The number of nucleotides increases the specificity of the probe and improves the Tm value, which is suitable for the characteristics of hMPV genome T / A abundance and low G / C content ( figure 1 ). Its combination is as follows:

[0009] Upstream primer: CATCAGGTAATATCCCACAAAATCAG

[0010] Downstream primer: GTGAATATTAAGGCACCTACACATAATAA

[0011] MGB probe: TCAGCACCAGACACAC

[0012] For the primer probe sequence of the L gene, the sequence homologous to the BJ1887 strain found in Beijing (its GenBank accession number is DQ843659) was selected. The original probe was a common TaqMan probe. When the method was established, it was found that the probe did not ampl...

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Abstract

The invention relates to the technical field of primer and probe sequences for detecting nucleonic acid of human metapneumovirus: a biotechnology and a virus detection technology and provides a primer and MGB (Myohaemoglobin) probe sequence for real-time RT-PCR (Reverse Transcription-Polymerase Chain Reaction) detection of human metapneumovirus nucleonic acid. The sequence is a primer and probe sequence which is designed and synthesized by carrying out sequence detection and analysis on an N gene and an L gene of known hMPV (human Metapneumovirus) and takes the L gene as a target. The sequence is researched and verified by 200 clinical samples detected with the real-time RT-PCR method, and the degenerate base group of the sequence includes the variation rule of 4 genetic subtypes of the hMPV, therefore, the sequence can be used for the research and the development of detection technology of nucleonic acid infected by the hMPV.

Description

technical field [0001] The invention relates to primers and probe sequences for real-time RT-PCR detection of human metapneumovirus nucleic acid. Background technique [0002] Human metapneumovirus (hMPV) is a single-stranded negative-sense RNA virus belonging to the subfamily Pneumoviridae of the family Paramyxoviridae. It is a new human respiratory virus discovered in 2001 by Dutch scholar Vanden Hoogen et al. Based on genome sequencing and systematic Evolutionary analysis shows that hMPV has two genotypes A and B, and the clinical symptoms and signs caused mainly include runny nose, cough, sore throat, fever, bronchitis, pneumonia, conjunctivitis, otitis media, etc., infants, the elderly and immunocompromised persons for susceptible populations. After the discovery of hMPV, there have been reports of hMPV infection in Europe, Australia, America, Asia and other countries, suggesting that the virus is a widespread human pathogen in the world. At the same time, the detecti...

Claims

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Application Information

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IPC IPC(8): C12Q1/70C12Q1/68C12N15/11
Inventor 崔丽瑾谭文杰陆柔剑张陵林周为民阮力
Owner 中国疾病预防控制中心病毒病预防控制所
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