Rapid detection method for bacillus cereus vomitoxin in food
A technology of Bacillus cereus and deoxynivalenol, which is applied in the field of microbial detection, can solve the problems of large matrix effect, complicated pretreatment, and inaccuracy, and achieve the effect of strong pertinence, high sensitivity, and strong specificity
Pending Publication Date: 2022-03-18
GUANGZHOU INST FOR FOOD INSPECTION(GUANGZHOU INSPECTION CENT FOR WINE & SPIRITS)
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[0003] At present, the detection methods of Bacillus cereus vomitoxin mainly include: 1. PCR method: distinguish by measuring the toxin-producing Ces gene, and it is impossible to determine whether there is toxin production; 2. Chemical detection method: mainly liquid chromatography-mass spectrometry and methods such as liquid chromatography ion trap mass spectrometry, the measurement of the production amount of Bacillus cereus toxin-producing strain Cereulide, the pretreatment is complicated, and the matrix effect is large; High; 4. Biological detection method: there are cell vacuolar response test method and in vitro sperm motility test method, only semi-quantitative detection of Cereulide, and it is difficult for general laboratories to carry out
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[0029] A rapid detection method for Bacillus cereus vomitoxin in food. In the present embodiment, artificially polluted rice with vomitoxin-producing Bacillus cereus is used as a sample for detection, which includes the following steps,
[0030] Step 1. Weigh an appropriate amount of sample, and obtain typical colonies after inoculation and cultivation;
[0031] Specifically: Weigh 25g of the sample, add 225mL of normal saline to homogenize, and use it as a 1:10 sample homogeneous solution; absorb 1mL of the 1:10 sample homogeneous solution, add 9mL of normal saline, and mix well to make a 1:100 sample homogeneous solution. According to the estimation of the contamination of the sample, according to the above operation, a ten-fold incremental serial dilution of the homogeneous sample was made sequentially; according to the estimation of the contamination of the sample, select 10 -1 、10 -2 、10 -3 These 3 sample homogeneous solutions with appropriate dilutions, each sample hom...
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The invention provides a method for rapidly detecting bacillus cereus vomitoxin in food. The method comprises the following steps: 1, weighing a sample to obtain a typical bacterial colony; 2, MALDI-TOF MS (Matrix Assisted Laser 2.1, sample application; respectively dispensing 1 microliter of typical bacterial colony, 1 microliter of PCS II standard solution and 1 microliter of Cereulide standard solution on the micropores of the target plate to form sample application micropores, drying liquid drops, and sequentially covering 1 microliter of 70% formic acid and 1 microliter of HCCA matrix solution for detection; 2.2, setting parameters; 2.3, correcting the instrument; 2.4, data acquisition and analysis; and analyzing by using flexanalysis software, and if two characteristic peaks of m / z1175 and m / z1191 exist, the result is positive, which indicates that the sample contains vomitoxin. Based on a matrix-assisted laser desorption / ionization time-of-flight mass spectrometry analysis method, the strain obtained after culture is directly analyzed, the vomitoxin is qualitatively identified, and the method has the characteristics of rapidness, high efficiency, high sensitivity and strong specificity.
Description
technical field [0001] The invention relates to the technical field of microbial detection, in particular to a rapid detection method for Bacillus cereus vomitoxin in food. Background technique [0002] Bacillus cereus vomitoxin (Cereulide, referred to as vomitoxin) is a circular small molecular peptide synthesized by the Ces gene encoding NRPS located on the giant plasmid, with a molecular weight of 1153.38Da, containing L-O-Val-L-Val-D-O- Leu-D-Ala three amino acid repeat sequence. The toxin has no antigenicity, is heat-resistant and acid-resistant, and is still active under the conditions of 126°C, 90min, and pH 2-12. Time exposure to high temperature and pressure environment can inactivate Cereulide, which is a common toxin that causes food poisoning. The toxin is pre-produced in food and is very stable. After entering the body, it binds to its receptors in the stomach, causing vomiting. The study found that there is a certain detection rate in ready-to-eat meat produ...
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CPCG01N27/64
Inventor 陈楷黄志深林秀敏宋安华肖剑雷燕劳嘉倩曹霞飞蒋佳希
Owner GUANGZHOU INST FOR FOOD INSPECTION(GUANGZHOU INSPECTION CENT FOR WINE & SPIRITS)