Method of detecting nucleotides of Bacillus cereus, primers for detecting and probes

A technology for detecting primers for Bacillus cereus, applied in the direction of microorganism-based methods, biochemical equipment and methods, microorganisms, etc., to achieve high sensitivity, high accuracy, and good primer specificity

Active Publication Date: 2014-12-03
SOUTH CHINA UNIV OF TECH
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  • Abstract
  • Description
  • Claims
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Problems solved by technology

Spore surface antigens are the basis for rapid immunoassays, but serological tests are largely affected by cross-reactivity of polyclonal and monoclonal antibodies with other antigens

Method used

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  • Method of detecting nucleotides of Bacillus cereus, primers for detecting and probes
  • Method of detecting nucleotides of Bacillus cereus, primers for detecting and probes
  • Method of detecting nucleotides of Bacillus cereus, primers for detecting and probes

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Embodiment 1

[0030] A method for detecting bacillus cereus, carried out according to the following steps:

[0031] 1. Design and synthesis of primers and probes

[0032] Download all Bacillus cereus genome sequences from the NCBI database, and select a highly conserved segment SEQ ID NO.4 with no secondary structure through comparative analysis of the sequences, and use the biological software PrimerExpress3.0 to design primers and Taqman probe. And the Blast homology comparison verification was carried out through the NCBI database to ensure that the sequence has no crossover with other species. The designed primer sequences are:

[0033] Forward primer: 5'-CATTCGTTCGCTGTGTGAACA-3'

[0034] Reverse primer: 5'-CCAACTTCAACCACAGGACAAA-3'

[0035] The sequence of the probe is: 5'-AGCCACCTAGCACACACGCCCAGA-3'

[0036] The 5' end of the probe was labeled with the reporter fluorescent dye VIC, and the 3' end was labeled with the quencher fluorescent dye BHQ2.

[0037] 2. Extraction of DNA ...

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Abstract

The invention discloses a method of detecting nucleotides of Bacillus cereus, primers for detecting and probes, belonging to the technical field of biological detection. According to the invention, the primers designed according to genome sequence of the Bacillus cereus are good in singularity, and high in sensitivity if used for fluorescence PCR detection. The detection method provided by the invention is high in accuracy and sensitivity, can rapidly and simply determine whether samples contain the Bacillus cereus, and provides scientific basis for etiological diagnosis and differential diagnosis of the Bacillus cereus in various dairy products.

Description

technical field [0001] The invention belongs to the technical field of biological detection, in particular to a method for detecting nucleotides of bacillus cereus, primers and probes for detection. Background technique [0002] Bacillus cereus is an aerobic, mesophilic, spore-forming, Gram-positive bacillus widely distributed in soil, water and air dust. There are many types of strains, including probiotic strains, low-toxicity strains and a few high-virulence strains. High-virulence strains often lead to food poisoning outbreaks and are common food-contaminating bacteria. Bacterial cells are rod-shaped, with square ends, in short or long chains, 1.0-1.2×3.0-5.0 microns. Produce spores, spores are round or cylindrical, mesozoic or near mesozoic, and cysts have no obvious swelling. Acapsular, motile. The colonies are large, rough, flat, and irregular. Bacillus cereus, Bacillus thuringiensis and Bacillus anthracis form the group of Bacillus cereus. Their morphological cha...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68C12N15/11C12R1/085
Inventor 肖性龙余以刚吴晖
Owner SOUTH CHINA UNIV OF TECH
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