Primer and method for detecting GI type and GII type norovirus by utilizing primer

A virus and primer sequence technology, applied in the direction of microorganism-based methods, biochemical equipment and methods, microorganism measurement/testing, etc., can solve the problem of low detection sensitivity and achieve simplified operation steps, shortened detection time, and high specificity Effect

Inactive Publication Date: 2012-01-04
湖州市疾病预防控制中心
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Problems solved by technology

Electron microscopy has the advantages of directness and reliability, but the detection sensitivity is low, and the virus titer of stool samples usually reaches 10 6 / g to meet the detection requirements

Method used

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  • Primer and method for detecting GI type and GII type norovirus by utilizing primer
  • Primer and method for detecting GI type and GII type norovirus by utilizing primer
  • Primer and method for detecting GI type and GII type norovirus by utilizing primer

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Embodiment

[0027] 1 Materials and methods

[0028] 1.1 Viruses

[0029] Rotavirus, adenovirus, and hepatitis A virus nucleic acids were donated to the Institute of Virology, Zhejiang Provincial Center for Disease Control and Prevention. Known Norovirus GⅠ and GⅡ positive strains are preserved in our laboratory. The clinical samples were fecal samples collected from patients during the 2008-2010 outbreak of non-bacterial acute gastroenteritis in Huzhou area.

[0030] 1.2 Primer design

[0031] The complete gene sequences of 60 strains of GⅠ and GⅡ norovirus strains from different years and regions were retrieved from the GenBank gene bank, and the homology comparison was carried out with biological software, and specific primers were designed in the conserved region of the capsid protein. for:

[0032] Nov-F: 5-GTGAATGAWGATGGCGTCKA-3 (nt5353-nt5372)

[0033] Nov-R: 5-GTHAGWATCCAGGGGTCAAT-3 (nt5528-nt5510)

[0034] The primers were synthesized by Shanghai Yingjun Biotechnology Co., L...

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Abstract

The invention relates to a primer and a method for detecting GI type and GII type norovirus by utilizing the primer, belonging to the technical field of biological detection. The sequence of the primer is as follows: Nov-F: 5-GTGAATGAWGATGGCGTCKA-3, Nov-R: 5-GTHAGWATCCAGGGGTCAAT-3. The invention focuses on the design of the sequence of the primer, the composition of an RT-PCR (reverse transcription-polymerase chain reaction) system, the selection of reaction conditions and the judgment of reaction results can be performed according to the conventional methods in the field. By adopting the primer and the method provided by the invention, a pair of primers can simultaneously detect and differentiate the GI type and the GII type norovirus in a same reaction tube, thereby not only simplifying the operation steps, but also shortening the detection time.

Description

technical field [0001] The invention relates to a primer and a method for detecting GⅠ and GⅡ norovirus using the primer, which belongs to the technical field of biological detection. Background technique [0002] Noroviruses (NVs), belonging to the Norovirus genus of the Caliciviridae family, are important pathogens that cause outbreaks and sporadic nonbacterial gastroenteritis worldwide. At present, norovirus can be divided into five genetic groups (GenogroupⅠ~Ⅴ, GⅠ~Ⅴ) according to the nucleotide sequence differences of the viral gene RNA polymerase (RNA dependent RNA polymerase, RdRp) and the main capsid protein VP1. Humans are mainly infected with GⅠ and GⅡ noroviruses, and each genetic group can be further divided into several genotypes. Because the virus has strong resistance, low pathogenic dose, and people of all ages are generally susceptible to it, it can often cause large-scale outbreaks in some public places such as schools, kindergartens, hospitals, etc. throug...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/11C12Q1/70C12Q1/68C12R1/93
Inventor 韩建康张红河吴晓芳纪蕾徐德顺汤仁树
Owner 湖州市疾病预防控制中心
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