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Detection kit, detection method and application for bacillus cereus

A Bacillus cereus, detection kit technology, applied in biochemical equipment and methods, microbial determination/inspection and other directions, can solve the problems of cumbersome operation, long detection time, high cost, simple operation, short reaction time, Easy-to-use effects

Active Publication Date: 2020-04-14
INSPECTION & QUARANTINE TESTING CENT OF HEBEI ENTRY EXIT INSPECTION & QUARANTINE BUREAU
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  • Abstract
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  • Claims
  • Application Information

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Problems solved by technology

[0005] Aiming at the problems of cumbersome operation, long detection time and high cost of the existing detection methods for Bacillus cereus, the present invention provides a detection kit for Bacillus cereus

Method used

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  • Detection kit, detection method and application for bacillus cereus
  • Detection kit, detection method and application for bacillus cereus
  • Detection kit, detection method and application for bacillus cereus

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Embodiment 1

[0046] Example 1 Detection method of Bacillus cereus-establishment of real-time fluorescent RPA method

[0047] 1. Design and preparation of primers and probe sequences

[0048] The inventors of the present invention analyzed the 16sRNA gene sequence (accession number: KY224970.1) of Bacillus cereus in GeneBank, selected the conserved region, and designed real-time fluorescent RPA, that is, real-time RPA primers and exo probes. The size of the target fragment is 297 bp.

[0049] The present invention designed and synthesized a pair of primers and a probe, as shown in Table 1.

[0050] Table 1 Bacillus cereus real-time RPA primers and probes designed by the present invention

[0051]

[0052] 2. Extraction of Bacillus cereus strains and bacterial genomic DNA

[0053] All Bacillus cereus used in this study were preserved by our laboratory.

[0054] Inoculate the pure culture of Bacillus cereus (CICC63301) into 10 mL of nutrient broth and incubate at 37°C for 16 hours; tak...

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Abstract

The invention relates to the field of biotechnology, and concretely discloses a detection kit as well as a detection method and application for bacillus cereus. The detection kit is used for detecting the bacillus cereus through real-time fluorescence RPA, and comprises a pair of primers based on a bacillus cereus 16sRNA gene sequence and a probe. The detection method has excellent specificity and sensitivity, the detection limit is 1.0 x 10<-3>ng / muL, and when the pollution load is larger than or equal to 1.5 x 10<4>CFU / g, the bacillus cereus can be detected through the real-time fluorescence RPA, and only 6 to 13min is required. Therefore, the real-time fluorescence RPA established by the invention can be used for quickly, efficiently and sensitively detecting the bacillus cereus, and provides a referential technological means for detection of the bacillus cereus in food processing quality control, food-borne epidemic diseases and other public health events.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a detection kit, detection method and application for bacillus cereus. Background technique [0002] As an opportunistic pathogen, Bacillus cereus is one of the main causes of food poisoning and can cause clinical symptoms characterized by nausea, vomiting, abdominal pain and diarrhea. The bacteria are widely distributed in soil, sewage and air, and are also common in various raw or cooked animal and plant foods. Bacillus cereus has the characteristics of diversified metabolic pathways, and has ecological adaptability and strong survival ability in different environments, which can resist adverse factors and spread better. Food is easily contaminated by Bacillus cereus due to irregular processing or improper storage. After the contaminated Bacillus cereus or remaining spores grow and multiply, the contaminated food has no obvious change in appearance and properties and is eaten by m...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6844C12Q1/04
CPCC12Q1/6844C12Q2531/119C12Q2521/507C12Q2537/1376C12Q2522/101C12Q2561/113
Inventor 王建昌刘立兵孙晓霞娄巧哲南汇珠
Owner INSPECTION & QUARANTINE TESTING CENT OF HEBEI ENTRY EXIT INSPECTION & QUARANTINE BUREAU
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