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A nucleic acid detection method for rapid identification of Bacillus cereus and Bacillus thuringiensis

A technology of Bacillus cereus and Bacillus cereus, applied in the field of microbial detection, can solve the problems of Bacillus thuringiensis and Bacillus cereus detection and failure to provide, and achieve fast detection speed, improved specificity, and wide credible range Effect

Active Publication Date: 2021-12-21
GUANGDONG INST OF MICROBIOLOGY GUANGDONG DETECTION CENT OF MICROBIOLOGY
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, this target failed to detect Bacillus thuringiensis and Bacillus cereus at the same time, and failed to provide a more intuitive and visual method

Method used

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  • A nucleic acid detection method for rapid identification of Bacillus cereus and Bacillus thuringiensis
  • A nucleic acid detection method for rapid identification of Bacillus cereus and Bacillus thuringiensis
  • A nucleic acid detection method for rapid identification of Bacillus cereus and Bacillus thuringiensis

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Embodiment 1

[0032] 1. Based on the genomes of the Bacillus cereus group or related species in GenBank and the existing Bacillus cereus and Bacillus thuringiensis, download the genome, remove redundant genome data that does not meet the analysis requirements (contigs ≥ 200), and modify the genome Name, using Prokka software for genome annotation; confirmatory analysis of the authenticity of genome taxonomic information of Bacillus cereus strains: average nucleotide homology analysis (ANI), based on existing international current Bacillus cereus or related species 25 reference bacterial strains (table 1) are used as the standard bacterial strain of Bacillus cereus group, and the similarity of the whole genome of the unknown bacterial strain and the standard bacterial strain is compared and analyzed, and the one with the highest similarity with the above-mentioned standard bacterial strain is selected, and the unknown bacterial strain is preliminarily considered to be this species; application...

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Abstract

The invention discloses a nucleic acid detection method for rapidly identifying bacillus cereus and bacillus thuringiensis. The present invention collects as many public databases as possible and the genomes of the Bacillus cereus group strains in the previous research, conducts authenticity analysis on the existing strain classification information, compares genomes and pan-genome methods to mine Bacillus cereus (Bacillus cereus) in the public database ) and Bacillus thuringiensis (Bacillus thuringiensis) can distinguish targets, using this group of experimental strains to verify and screen the characteristic molecular targets that distinguish Bacillus cereus and Bacillus thuringiensis, including selecting different SNP site designs in the molecular target ispD gene sequence For the screening of primers and primer pairs, HRM experiments were carried out with Bacillus cereus and Bacillus thuringiensis as templates respectively, and the characteristic melting curve information of the corresponding strains was obtained to realize the accurate identification of Bacillus cereus and Bacillus thuringiensis.

Description

Technical field: [0001] The invention belongs to the field of microorganism detection, and in particular relates to a nucleic acid detection method for rapidly identifying Bacillus cereus and Bacillus thuringiensis through high-resolution melting curves. Background technique: [0002] Bacillus cereus (Bacillus cereus) and Bacillus thuringiensis (Bacillus thuringiensis) belong to the Bacillus cereus group. Among them, Bacillus cereus is a common foodborne opportunistic pathogen in my country, which can produce diarrhea (enterotoxin) toxin and vomitoxin. From 2008 to 2015, Bacillus cereus ranked third in the prevalence of foodborne pathogens in my country, and it is highly harmful. Bacillus thuringiensis is a class of insect pathogenic bacteria. When it forms spores, it will form one or more parasporal crystals, and its main component is insecticidal crystal protein, so it is widely used as a biopesticide. The physiological and biochemical characteristics of Bacillus cereus...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/689C12Q1/686C12Q1/04C12N15/11C12R1/085C12R1/07
CPCC12Q1/689C12Q1/686C12Q2600/156C12Q2527/107
Inventor 丁郁张俊卉周桓吴清平王涓朱振军张菊梅叶青华陈谋通薛亮吴诗曾海燕庞锐张淑红杨小鹃
Owner GUANGDONG INST OF MICROBIOLOGY GUANGDONG DETECTION CENT OF MICROBIOLOGY