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Detection of enterotoxigenicEscherichia coliin water by polymerase chain reaction amplification and hybridization

 

作者: Z. Tamanai-Shacoori,   A. Jolivet-Gougeon,   M. Pommepuy,   M. Cormier,   R. R. Colwell,  

 

期刊: Canadian Journal of Microbiology  (NRC Available online 1994)
卷期: Volume 40, issue 4  

页码: 243-249

 

ISSN:0008-4166

 

年代: 1994

 

DOI:10.1139/m94-040

 

出版商: NRC Research Press

 

数据来源: NRC

 

摘要:

EnterotoxigenicEscherichia coliwas studied in waste water, river water, and seawater from six locations along the west coast of Normandy by using the polymerase chain reaction (PCR) to amplify the heat labile (LT) gene. Cellular DNA was extracted from centrifugation pellets and amplified using PCR. The PCR products were detected by gel electrophoresis and confirmed by hybridization assay, using an 850 base pairHindIII DNA fragment probe from pEWD299 conjugated to digoxigenin and specific for the LT gene. Results of the PCR amplification were compared with those of GM1 enzyme-linked immunosorbent assay, latex agglutination, and colony hybridization. The PCR method was found to be more precise and less time consuming, especially when compared with methods requiring culture of isolates for enumeration of enterotoxigenicE.coliin water.Key words: enterotoxigenicEscherichia coli, PCR, environmental water, digoxigenin.

 

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