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Bacillus cereus detection method and kit

A technology for the detection of Bacillus cereus, which is applied in the field of rapid detection of Bacillus cereus, can solve the problems of low specificity, cumbersome operation, and difficulty in identification, and achieve the effect of rapid detection method, high sensitivity, and strong specificity

Active Publication Date: 2014-06-11
SHANGHAI JIAOTONG UNIV +1
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Problems solved by technology

[0005] However, traditional detection methods are time-consuming and cumbersome to operate, while products based on immunological principles have problems such as high detection limits and low specificity
Especially the 6 species of Bacillus cereus, including B.cereus, B.anthracis, B.thuringiensis, B.mycoides , B.pseudomycoides and B.weihenstephanensis, the phenotype and biochemical characteristics are very similar, and there are only differences in the types of enterotoxins and the presence or absence of parasporal crystals. In particular, the existence of some mutant strains that do not produce enterotoxins or carry parasporal crystals makes the identification of this group very difficult

Method used

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  • Bacillus cereus detection method and kit
  • Bacillus cereus detection method and kit
  • Bacillus cereus detection method and kit

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Embodiment Construction

[0026] A preferred embodiment of the present invention is described in detail below with reference to the accompanying drawings.

[0027] Hereinafter, the invention will be described through specific examples. If not otherwise specified, according to the experimental manuals such as "Molecular Cloning Experiment Guide" (Third Edition) (Science Press, Beijing, China, 2005) familiar to those skilled in the art and the references cited herein The methods listed are implemented.

[0028] 1.1 Main reagents and instruments

[0029] The QIAamp DNA Mini Kit kit mentioned in the following description was purchased from Qiagen, USA (Cat.No.51304); Taqman Universal PCR Master Mix II with UNG fluorescent quantitative PCR Kit was purchased from Invitrogen, USA (Cat.No.4440038) ; ABI7500 fluorescent quantitative PCR instrument, purchased from ABI company.

[0030] 1.2 Extraction of strains and DNA

[0031] The strains can be purchased from the China Agricultural Microorganism Culture Coll...

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Abstract

The present invention provides a new Bacillus cereus detection method, which comprises: 1) extracting DNA from a sample requiring detection; and 2) adopting the DNA obtained from the step 1) as a template to carry out real-time fluorescence quantitative PCR detection, wherein a target detected by the real-time fluorescence quantitative PCR detection is Bacillus cereus murB gene. The present invention further provides a Bacillus cereus detection kit, which comprises primers for Bacillus cereus murB gene amplification.

Description

technical field [0001] The invention belongs to nucleic acid detection in the field of food safety. Specifically, the present invention relates to a method and a kit for rapid detection of Bacillus cereus. Background technique [0002] Bacillus cereus, as one of the aerobic bacillus group, is a non-capsulated, motile, Gram-positive bacillus, and its spores can withstand 100°C for 30 minutes. The bacterium is widely distributed in nature and is commonly found in soil, sewage and dust; it can grow and reproduce at 16-45°C, and the optimum growth temperature is 35°C. Food poisoning is often caused by the bacteria contaminating foods such as carbohydrate-rich starch products and protein-rich dairy products and growing and multiplying in them. Bacillus cereus is also an opportunistic pathogen of various non-intestinal infectious diseases in humans and animals, which can cause conjunctivitis, respiratory system, cardiovascular system, central nervous system, wound infection and ...

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68C12R1/085
Inventor 施春雷杨捷琳史贤明刘斌潘良文袁辰刚
Owner SHANGHAI JIAOTONG UNIV
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