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Primer for coxsackie virus A16 nucleic acid detection, probe and kit

A coxsackie virus, A16 technology, applied in the field of probes and kits, coxsackie virus A16 nucleic acid detection primers, can solve the problems of long detection cycle, prone to missed detection, cumbersome steps, etc., and achieve high accuracy , high sensitivity and good primer specificity

Inactive Publication Date: 2010-03-24
何雅青 +1
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  • Application Information

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Problems solved by technology

The classic methods for detecting enteroviruses mostly use cell culture and specific antibody detection. This method has cumbersome steps and a long detection cycle, and some enteroviruses are difficult to proliferate in cells, and are prone to missed detection; traditional RT-PCR is also used Used to detect EV71 and CA16, but the detection time takes 6 hours

Method used

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  • Primer for coxsackie virus A16 nucleic acid detection, probe and kit
  • Primer for coxsackie virus A16 nucleic acid detection, probe and kit
  • Primer for coxsackie virus A16 nucleic acid detection, probe and kit

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Embodiment Construction

[0020] The following examples are used to further illustrate the present invention, but are not intended to limit the scope of the present invention.

[0021] Primer and probe design and synthesis :

[0022] According to the CA16 VP1 gene sequence in GenBank, primers and Taqman probes were designed at its conserved sites using the biological software Primer Express V2.0, and Blast homology comparison verification was carried out through the NCBI database. The primer sequences are:

[0023] CA16Pf (Forward): 5'-GAACCATCACTCCACACAGGAG-3'

[0024] CA16Pr (reverse): 5'-GTACCCGTGGTGGGCATTG-3'

[0025] The sequence of the probe is: 5'-CAGCCATTGGGAATTTCTTTAGCCGTG-3'

[0026] The 5' end of the probe was labeled with the reporter fluorescent dye VIC, and the 3' end was labeled with the quencher fluorescent dye BHQ2.

[0027] RNA extraction :

[0028] Extract RNA from virus samples with Roche High Pure viral RNA kit, take about 0.5 g of feces samples, add 500 μl of TE to make a...

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Abstract

The invention relates to a primer for coxsackie virus A16 nucleic acid detection, probe and kit, wherein the nucleotide sequence of the primer is that: forward primer 5'-GAACCATCACTCCACACAGGAG-3'; backward primer 5'-GTACCCGTGGTGGGCATTG-3' and the nucleotide sequence of the probe is 5'-CAGCCATTGGGAATTTCTTTAGCCGTG-3'. The invention also provides a method for detecting coxsackie virus A16 nucleic acid. Sample RNA is used as template and the above primer and probe are subjected to real-time fluorescence RT-PCR amplification and the result is predicated based on the amplification curve after each circulation. The primer specificity is good, the detection method is quick and simple, the accuracy and the sensitivity is high and the invention provides scientific reference for etiologic diagnosis and differential diagnosis of hand-foot-mouth disease and relative diseases.

Description

technical field [0001] The invention relates to biological detection technology, in particular to primers, probes and kits for coxsackievirus A16 nucleic acid detection. Background technique [0002] Coxsackievirus A16 (CA16 for short) is a single-stranded positive-sense RNA virus belonging to the Picoranviridae family. CA16 is an important pathogen that causes hand, foot and mouth disease in children. The clinical symptoms caused by it are difficult to distinguish from hand, foot and mouth disease caused by enterovirus 71 infection. However, under normal circumstances, CA16 infection generally does not cause serious neurological related diseases. , is often overlooked due to its subclinical infection. Many countries and regions in the world have reported the prevalence of hand, foot and mouth disease caused by CA16, and there is a pandemic every 3-5 years. In recent years, CA16 has been paid more and more attention due to its association with myocarditis, pericarditis and...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/70C12Q1/68C12N15/11
Inventor 何雅青肖性龙
Owner 何雅青
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