Direct PCR (Polymerase Chain Reaction) detection method of Bombyx mori nuclear polyhedrosis pathogeny BmNPV

Abstract

The invention discloses a direct PCR detection method of Bombyx mori nuclear polyhedrosis pathogeny BmNPV, comprising the following steps of: firstly, treating the blood of infected Bombyx mori by ethanol precipitation to remove a PCR reaction inhibitor; designing the PCR primer of a polyhedrin gene according to the genome analysis of the Bombyx mori nuclear polyhedrosis pathogeny BmNPV; carrying out PCR amplification on the polyhedrin gene, adding an anti-PCR reaction inhibitor BSA (Bull Serum Albumin) to a reaction system and simultaneously carrying out PCR reaction by the primer of a Bombyx mori chondriosome CO I gene to make positive control; respectively getting the blood of normal Bombyx mori and the inflected midguts of the Bombyx mori infected with white muscardine, green muscardine, a bacterial disease and a microsporidia disease and then carrying out PCR reaction specification analysis; and detecting the PCR reaction result by agar gel electrophoresis. The whole detection process has simple operation and can be completed only by 4 hours. The disease can be diagnosed at the early stage of BmNPV infection by adopting the direct PCR detection method to carry out periodical sampling detection so as to adopt a measure in time to prevent the disease from happening.

Description

[0001] technical field [0002] The invention relates to early molecular biological diagnosis technology of silkworm blood type pus disease caused by silkworm nucleopolyhedrosis virus BmNPV, and belongs to the biological technology field of silkworm disease prevention and treatment. Background technique [0003] The silkworm is a Lepidoptera insect with important economic value. In the international market, my country's raw silk exports account for more than 90% of the world's total raw silk exports, silk exports account for more than 50%, and silk clothing exports also account for a considerable proportion. [0004] Bombyx mori nuclear polyhedrosis virus (BmNPV) is a circular double-stranded DNA. The silkworm nuclear polyhedrosis caused by it is the most important disease in cocoon silk production. The disease is highly contagious and difficult to control. Every year, the loss of sericulture production in China due to silkworm disease accounts for 30% of the total output, o...

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Problems solved by technology

Hybridization can detect 20-5000 PIBs (0.1ngDNA), but the technical operation of this method is very complicated, which seriously limits its application in disease diagnosis

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