Fluorescent quantitative PCR (polymerase chain reaction) detection method for bee paenibacillus larvae TaqMan

Abstract

The invention belongs to the field of detection technologies for bee paenibacillus larvae, and discloses a fluorescent quantitative PCR (polymerase chain reaction) detection method for bee paenibacillus larvae TaqMan. The fluorescent quantitative PCR detection method has the advantages that a pair of specific primers and a TaqMan fluorescent probe are designed according to P. larvae 16S rRNA [ribosomal RNA (ribonucleic acid)] gene sequences, the fluorescent quantitative PCR detection method for bee American foulbrood pathogenic bacteria TaqMan is created, and accordingly the shortcomings in the prior art can be overcome; the created fluorescent quantitative PCR detection method is good in specificity and repeatability and high in sensitivity, and variation coefficients of Ct values of intra-assay and inter-assay repetitive tests are lower than 3%; the fluorescent quantitative PCR detection method is wide in application range, results obtained by the aid of the fluorescent quantitative PCR detection method for detecting laboratory simulated samples of adult bees, larvae, bee pupae, honey, propolis, royal jelly, pollen and the like are consistent with expected results, and accordingly the fluorescent quantitative PCR detection method can be used for early and high-throughput diagnosis on AFB (acid-fast bacilli) in honey and bee products and quantitative analysis on P. larvae.

Description

technical field [0001] The invention belongs to the technical field of honeybee larval bacillus detection, in particular to a TaqMan fluorescent quantitative PCR detection method for honeybee larval bacillus. Background technique [0002] American foulbrood (AFB) is a bacterial, acute, and devastating infectious disease of bee larvae and pupae caused by Paenibacillus larvae (P.larvae), and its typical symptoms are death after capping The larvae's nest cover becomes damp, sunken, darkened and oily, and most of the nest cover is perforated. Picking the larvae can pull out colloidal filaments with a length of 10cm to 15cm. The diseased larvae become soft and lose their normal White and glossy, gradually turning light brown to brownish black. This bacillus is surrounded by a 7-layer structure. The special structure makes it extremely viable and can survive for more than 35 years in harsh environments. Once AFB occurs in the bee farm, it is very easy to cause the death of larva...

AI Technical Summary

Problems solved by technology

[0004] 1. Microfluidic chip quantitative PCR detection speed is fast, but the detection cost is high, which is not conducive to popularization and application at present;

[0005] 2. The detection cost of SYBR Green fluorescent quantitative PCR is low, but the false positive rate of detection results is high and the specificity is poor

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