LAMP primer combination for detecting Bombyx mori nuclear polyhedrosis virus and kit

A nuclear polyhedron and kit technology, applied in the direction of recombinant DNA technology, microbial measurement/inspection, biochemical equipment and methods, etc., can solve the problems that do not reflect the improvement of amplification efficiency, and do not involve primer screening, etc., to achieve The effect of simple early diagnosis and avoidance of pollution

Inactive Publication Date: 2016-07-27
GUANGXI UNIV
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Problems solved by technology

Although the LAMP detection technology of BmNPV has also been reported, it does not involve the screening of primers and does not reflect the improvement of amplification efficiency

Method used

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  • LAMP primer combination for detecting Bombyx mori nuclear polyhedrosis virus and kit
  • LAMP primer combination for detecting Bombyx mori nuclear polyhedrosis virus and kit
  • LAMP primer combination for detecting Bombyx mori nuclear polyhedrosis virus and kit

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Embodiment Construction

[0030] The polh gene of the silkworm nuclear polyhedrosis virus is a structural protein gene overexpressed in the very late stage of the virus, and the encoded polyhedrosis protein assembles to form the polyhedron crystal structure of the virus. The inventor compared the polh gene sequence information of 45 Guangxi BmNPV strains in the early stage, and the results showed that the homology was greater than 99.5%, showing high conservation, so the polh gene was selected as the target gene to design LAMP primers. Taking the polh gene of the silkworm nuclear polyhedrosis virus as the detection target, according to the complete open reading frame sequence of the gene, the optimized primers were designed, synthesized and screened, and the corresponding LAMP detection kit was prepared. The specific research experimental steps are as follows:

[0031] 1. Material preparation

[0032]Bombyx mori nuclear polyhedrosis virus, silkworm cytoplasmic polyhedrosis virus, silkworm microsporidi...

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Abstract

The invention discloses an LAMP primer combination for detecting Bombyx mori nuclear polyhedrosis virus and a kit. The primer combination comprises external primers, internal primers and a loop primer, wherein the external primers F3 and B3, the internal primers FIP and BIP and the loop primer LF have base sequences of a sequence list from SEQ.ID.No.1 to SEQ.ID.No.5. The primer combination is contained in the kit. By means of the LAMP primer combination for detecting Bombyx mori nuclear polyhedrosis virus and the kit, Bombyx mori nuclear polyhedrosis virus can be detected only by performing DNA extraction and loop-mediated isothermal amplification on a tissue sample, thus early diagnosis of Bombyx mori nuclear polyhedrosis virus in a production field can be carried out quickly, accurately and easily. Compared with the prior art, the LAMP primer combination for detecting Bombyx mori nuclear polyhedrosis virus and the kit have the advantages of high specificity, high sensitivity, quick result obtaining and no pollution.

Description

technical field [0001] The invention belongs to the technical field of silkworm nuclear polyhedrosis virus detection, in particular to a LAMP primer set and a kit for detecting the silkworm nuclear polyhedrosis virus. Background technique [0002] Bombyx mori blood type pus disease is the most common and most harmful viral infectious disease in sericulture. Due to strong infectivity, high case fatality rate, and currently no effective treatment, the epidemic of the disease causes huge economic losses to silkworm cocoon production every year. The pathogen of the disease is Bombyxmorinucleopolyhedrovirus (BmNPV), which is characterized in that the virus particles can preserve the infection activity for a long time under the protection of the polyhedron structure. The polyhedron is ingested by silkworms and causes primary infection, and then spreads to other healthy silkworms in the silkworm room. Under the joint action of human factors such as trade, traffic, and natural fact...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/70C12Q1/68C12N15/11
CPCC12Q1/6844C12Q1/701C12Q2531/119C12Q2563/107
Inventor 梁湘屈达才李俊陈慧珠卓秋红龙羽燕
Owner GUANGXI UNIV
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