Rapid detection kit and detection method for bombyx mori nuclear polyhydrosis virus

A karyotype polyhedron and detection kit technology, applied in biochemical equipment and methods, microbial determination/inspection, etc., can solve the problems of high cost, cumbersome operation, inability to detect latent virus samples, etc., and achieve detection consumption. The effect of short time and high detection sensitivity

Inactive Publication Date: 2015-03-25
JIANGSU UNIV
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Problems solved by technology

[0003] At present, the detection methods of the virus include electron microscope method, PCR detection method, etc., but in the actual detection work, the existing detection technology has the following problems: first, the sensitivity is not enough to detect latent virus-infected samples; second, the cost is too high, It is not suitable for promotion and use; the third is that the operation is cumbersome and requires high experimental conditions, which is not conducive to farmers' adoptio

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  • Rapid detection kit and detection method for bombyx mori nuclear polyhydrosis virus
  • Rapid detection kit and detection method for bombyx mori nuclear polyhydrosis virus
  • Rapid detection kit and detection method for bombyx mori nuclear polyhydrosis virus

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Embodiment 1

[0051] 1. Materials

[0052] The samples infected with silkworm nuclear polyhedrosis virus came from our laboratory; the primers and probes used were synthesized by Shanghai Sangon Biotechnology Company; the large fragment of Bst DNA polymerase was purchased from New England BIPolabs Company; dNTP was purchased from Promega Company; betaine, Magnesium sulfate, etc. were purchased from sigma company.

[0053] 2. Extraction of DNA from diseased silkworm tissues:

[0054] (1) Take 20mg of silkworm eggs, hemolymph or midgut tissue, add 200μL of lysate TL and grind with a disposable grinding rod. After thorough homogenization, add 20μL of proteinase K solution (20mg / ml), shake and mix immediately , and then placed in a 55°C water bath for 1 to 3 hours until the tissue is completely digested;

[0055] (2) Transfer the digestion solution to the adsorption column AC, place the adsorption column in the collection tube, centrifuge at 12000rpm for 1min, and discard the waste liquid in ...

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Abstract

The invention relates to a rapid detection kit and a detection method for bombyx mori nuclear polyhydrosis virus, specifically relates to a quick and convenient detection kit for detecting the bombyx mori nuclear polyhydrosis virus by combining the loop-mediated isothermal amplification technology and the nucleic acid detection lateral chromatography test paper, and a detection method of the detection kit, and belongs to the technical field of viral epidemic disease diagnosis. The detection kit is characterized by comprising a BmNPVLAMP pre-reaction liquid, wherein the desired primer and probe are designed according to the conserved region of the genomic sequence of the bombyx mori nuclear polyhydrosis virus published by GenBank; the kit is capable of effectively and quickly detecting the bombyx mori nuclear polyhydrosis virus; the steps of virus DNA extraction and BmNPVLAMP reaction system amplification are performed, and after the reaction ends, the result is judged by use of the quick nucleic acid detection test paper to obtain that the BmNPV virus DNA is efficiently and specifically amplified; the detection method is simple, convenient and quick, good in specificity, and high in sensitivity, and is suitable for farmers to use daily and advantageous for diagnosing and preventing the bombyx mori nuclear polyhydrosis virus disease.

Description

technical field [0001] The invention relates to a rapid detection kit and detection method for silkworm nuclear polyhedrosis virus, in particular to a loop-mediated isothermal amplification (Loop-mediated isothermal amplification, LAMP) technology and nucleic acid detection lateral flow chromatography test paper (nucleic acid Detection lateral flow dipstick (LFD) combined with a fast and convenient detection kit for detecting silkworm nuclear polyhedrosis virus and a detection method thereof, belong to the technical field of virus epidemic diagnosis. Background technique [0002] The silkworm is an important economic insect, and the silk it produces is the main source of silk raw materials, and plays an important role in human economic life and history and culture. However, the high incidence of silkworm disease directly affects the yield and quality of silkworm cocoons, restricting the healthy and sustainable development of sericulture. Silkworm diseases are divided into i...

Claims

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Application Information

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IPC IPC(8): C12Q1/70C12Q1/68
CPCC12Q1/6844C12Q1/70C12Q2531/119C12Q2565/625
Inventor 周阳施海峰高力陈亮刘海涛张春霞陈克平
Owner JIANGSU UNIV
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