Bifidobacterium breve and detection method of methamidopho pesticide residue in foodstuff

A technology of Bifidobacterium breve and a detection method, which is applied in the field of microbial detection of methamidophos pesticides, can solve problems such as the lack of sensitivity and specificity of methamidophos for screening, and achieves that the detection method is simple, easy, practical and stable. good effect

Inactive Publication Date: 2012-06-27
温州医科大学仁济学院
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  • Abstract
  • Description
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Problems solved by technology

So far, there is no relevant report on the screening of working strains of bifidobacteria with high sensitivity and specificity for methamidophos and its application in the detection of methamidophos residues in food

Method used

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  • Bifidobacterium breve and detection method of methamidopho pesticide residue in foodstuff
  • Bifidobacterium breve and detection method of methamidopho pesticide residue in foodstuff
  • Bifidobacterium breve and detection method of methamidopho pesticide residue in foodstuff

Examples

Experimental program
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Effect test

Embodiment 1

[0037] Example 1: Isolation and identification of Bifidobacterium breve LJM-006 strain

[0038] Taking the feces of a healthy young man in Zhejiang Province as an isolated sample, on MRS agar medium, under anaerobic conditions at 37°C, smear and isolate and culture for 48 hours to obtain Bifidobacterium breve LJM-006 according to the present invention, which was identified as Bifidobacterium breve, this strain has been stored in the General Microbiology Center of China Committee for Microorganism Culture Collection on October 28, 2011 (Address: Institute of Microbiology, Chinese Academy of Sciences, No. 3, No. 1, Beichen West Road, Chaoyang District, Beijing, postal code 100101) Preservation, classification name is Bifidobacterium breve (Bifidobacterium breve), preservation number is CGMCC No.5418.

[0039] The formula of MRS agar medium: peptone 10g, beef extract 10g, yeast extract 5g, glucose 20g, sodium acetate 5g, K 2 HPO 4 2g, MgSO 4 7 hours 2 O 0.5g, MnSO 4 4H 2 O...

Embodiment 2

[0051] Example 2: Detection of methamidophos residues in vegetable samples

[0052] (1) Strain activation: the above-mentioned Bifidobacterium breve LJM-006 strain was inoculated on a nutrient agar slant medium, and the pH value of the medium was adjusted to 6.8-7.2, and optimized culture was carried out at 35-37°C, and the subsequent transformation was carried out. subculture;

[0053] (2) Bacterial suspension preparation: inoculate the Bifidobacterium breve LJM-006 obtained in step (1) on a nutrient agar slant medium, culture anaerobically at 35±2° C. for 24 hours, wash the bacterial lawn with sterilized normal saline, Make a suspension and adjust its OD 600 When the value reaches 0.25~0.35, store it in a refrigerator at 2-8°C;

[0054] (3) Detection tube preparation: by weight, take peptone 10g, beef extract 10g, yeast extract 5g, glucose 20g, sodium acetate 5g, K 2 HPO 4 2g, MgSO 4 7 hours 2 O 0.5g, MnSO 4 4H 2 O 0.2g, fructo-oligosaccharide 3g, diammonium citrate...

Embodiment 3

[0079] Example 3: Detection of methamidophos residues in cucumber samples

[0080] The food sample that detects is cucumber, gets the cucumber homogenate tissue 5g to be tested, and methamidophos standard substance is added in 10 cucumber samples with final concentration as 100 mg / L, measures its content by above embodiment 2 steps, according to B / B of each sample 0 value on the standard curve to find its corresponding methamidophos mass concentration, or substitute it into the regression equation to calculate the logarithmic value X of the sample mass concentration, and find its antilog, which is the mass concentration of methamidophos contained in the sample, and obtain methylamine The measured value of phosphorus content was 117.26±0.79mg / L, and the recovery rate of standard addition was 82.3%~115.0%.

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Abstract

The invention relates to bifidobacterium breve and a method for detecting methamidopho pesticide residue in foodstuff by using bifidobacterium breve. According to the invention, a selected strain is an LJM-006 strain with a collection number of CGMCC No. 5418. The LJM-006 strain is hypersensitive to methamidopho. The methamidopho residue detection method established employing the strain as a working strain comprises the steps of: (1) strain activation, (2) strain suspension preparation, (3) detection tube preparation, (4) standard curve drawing and regression equation establishing, (5) sample pretreatment, (6) sample detection, and the like. The detection method provided by the invention is advantaged in simple operation, low cost, and high sensitivity. The result of detection is easy to determine. Compared to a high-efficiency liquid chromatography method, a result coincidence rate is high. Therefore, the method is suitable for sample detection. The method satisfies the requirement by the national pesticide residue detection department for large-batch sample screening.

Description

technical field [0001] The invention relates to the field of microbial detection of methamidophos pesticides, and relates to a bifidobacterium breve strain and a method for detecting methamidophos pesticide residues in food by using the strain as a working strain. Background technique [0002] Food is the material basis for the survival and development of human beings. Excessive pesticide residues have become one of the outstanding problems of food safety in my country. Since 2010 alone, a series of food safety incidents caused by pesticide residues have occurred in my country, such as "Poisonous cowpea in Hainan" and "1930 kg of leeks detected to exceed the standard in Qingdao". Therefore, the timely and accurate analysis and detection of pesticide residues in food is of great importance, and the establishment of fast, simple, safe and accurate pesticide residue detection technology is an inevitable demand for the development of food safety. [0003] Methamidophos is a broa...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12Q1/02G01N21/31C12R1/01
CPCC12N1/20C12R1/01C12P1/00G01N33/02C12Q1/10C12Q1/18C12N1/205C12R2001/01
Inventor 刘佳明杜季梅孙晶曹建明
Owner 温州医科大学仁济学院
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