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Rapid Detection and Biological Characteristic for an Important Food-borne Pathogen, Escherichia Coli O157

Author: ZhuQi
Tutor: FangWeiHuan；MengPing
School: Zhejiang University
Course: Veterinary
Keywords: E. coli O157:H7 SYBR Green standard curve Ct PCR biochemical identification
CLC: S852.61
Type: Master's thesis
Year: 2009
Downloads: 35
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Abstract


Escherichia coli 0157 is an important food-borne pathogen. Separation and identification of this bacterium and further research in molecular level could have important significance. The techniques of separation and identification of E. coli 0157 in laboratory, such as plate separation, immunomagnetic separation, and polymerase chain reaction were reviewed, and made a comparision of various methods.E. coli 0157 and Salmonella are the main pathogens in pork which caused diarrhoea, chills and fever disease, etc. for consumer. This study was aimed detect the comtamination by the two pathogens along the pork production. process. The samples of pork were daily collected from pig hair, body surface abdominal wall and the wall of slaughtering plant in Jiaxing for 20 days from March to April 2008. First, PCR was applied to detect E. coli O157 and Salmonella for 84 samples, the contaminated sample were identified through biochemistry technique. The results showed that E. coli 0157 contaminated rates were of approximately 32.1%(27/84), and Salmonella contaminated rates were of approximately 11.9%(10/84).As an important food-borne pathogens, E. coli 0157:H7 can lead to serious diarrhea and hemorrhagic colitis hemolytic uremic syndrome. Given that conventional detection of E. coli O157:H7 is ineffective, a sensitive and specific detection methad is necessary for detecting E. coli O157:H7 in foods. In this study, a rapid and accurate real-time quantitative polymerase chain reaction (Real-time PCR) method using SYBR Green I for detecting Escherichia coli O157:H7 was established. A pair of primers were designed to amplify rfbE gene. The melting curves showed that the amplified product was specific. The optimal reaction program and standard curve were founded, and the Ct value of each sample could get quantitative results according to the standard curve. The result indicated that the method was 1000 times more sensitive than regulor PCR, and can detect E. coli O157:H7 more quickly and effectively.

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CLC: > Agricultural Sciences > Livestock, animal medicine,hunting,silkworm,bee > Animal Medicine ( Veterinary Medicine) > Basic Veterinary Science > Animal Microbiology ( Veterinary Microbiology, ) > Pathogenic bacteria
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