ARS | Publication request: ISOLATION OF CAMPYLOBACTER AND IDENTIFICATION BY PCR

Submitted to: Methods in Molecular Biology
Publication Type: Book / Chapter
Publication Acceptance Date: December 20, 2001
Publication Date: November 20, 2002
Citation: Englen, M.D., Ladely, S.R., Cray, P.J. 2002. Isolation of campylobacter and identification by pcr. Methods in Molecular Biology. Vol. 7., P. 109-121.

Interpretive Summary: The volume "PCR Detection of Microbial Pathogens" is designed to show the practical possibilities and problems connected with the use of PCR- based detection of important animal pathogens, particularly zoonotic agents. It intends to give microbiologists, veterinarians, molecular biologists and biochemists the opportunity to extend their knowledge on state-of-the-art detection procedures, as well as pre-PCR sample processing and general aspects of PCR. Our chapter, "Isolation of Campylobacter and Identification by PCR", provides workers unfamiliar with Campylobacter culture and identification a useful set of methods to serve as a practical starting point. The chapter includes methods for isolation of Campylobacter from feces, extraction of DNA for use as template in the PCR reaction, and PCR assays specific for C. jejuni and C. coli. A brief section on agarose gel electrophoresis of the amplification products using ethidium bromide staining for detection has also been included.

Technical Abstract: Campylobacter species are common inhabitants of the intestinal tracts of poultry and livestock and have been associated with food products of animal origin. Campylobacter is now recognized as a major food borne pathogen, with C. jejuni and C. coli accounting for the majority of human infections. Consequently, more and more researchers and public health personnel are now working with these organisms. The following protocols are routinely used in our laboratory, and are intended to provide workers unfamiliar with Campylobacter culture and identification a useful set of methods to serve as a practical starting point. This chapter outlines methods for isolation of Campylobacter from feces, extraction of DNA for use as template in the PCR reaction, and PCR assays specific for C. jejuni and C. coli. A brief section on agarose gel electrophoresis of the amplification products using ethidium bromide staining for detection has also been included.