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Journal of Clinical Microbiology, April 2008, p. 1220-1225, Vol. 46, No. 4
0095-1137/08/$08.00+0     doi:10.1128/JCM.02158-07
Copyright © 2008, American Society for Microbiology. All Rights Reserved.

High-Resolution Genotyping of Campylobacter Species by Use of PCR and High-Throughput Mass Spectrometry{triangledown}

James C. Hannis,1 Sheri M. Manalili,1 Thomas A. Hall,1 Raymond Ranken,1 Neill White,1 Rangarajan Sampath,1 Lawrence B. Blyn,1 David J. Ecker,1 Robert E. Mandrell,2 Clifton K. Fagerquist,2 Anna H. Bates,2 William G. Miller,2 and Steven A. Hofstadler1*

Ibis Biosciences, a Subsidiary of ISIS Pharmaceuticals Inc., 1891 Rutherford Road, Carlsbad, California 92008,1 USDA, ARS, WRRC, Produce Safety and Microbiology Research Unit, Albany, California 947102

Received 7 November 2007/ Returned for modification 20 December 2007/ Accepted 5 February 2008

In this work we report on a high-throughput mass spectrometry-based technique for the rapid high-resolution identification of Campylobacter jejuni strain types. This method readily distinguishes C. jejuni from C. coli, has a resolving power comparable to that of multilocus sequence typing (MLST), is applicable to mixtures, and is highly automated. The strain typing approach is based on high-performance mass spectrometry, which "weighs" PCR amplicons with enough mass accuracy to unambiguously determine the base composition of each amplicon (i.e., the numbers of A's, G's, C's, and T's). Amplicons are derived from PCR primers which amplify short (<140-bp) regions of the housekeeping genes used by conventional MLST strategies. The results obtained with a challenge panel that comprised 25 strain types of C. jejuni and 25 strain types of C. coli are presented. These samples were parsed and resolved with demonstrated sensitivity down to 10 genomes/PCR from pure isolates.

* Corresponding author. Mailing address: Ibis Biosciences, Inc., 1891 Rutherford Road, Carlsbad, CA 92008. Phone: (760) 603-2599. Fax: (760) 603-4653. E-mail: shofstad{at}ibisbio.com

{triangledown} Published ahead of print on 13 February 2008.

Journal of Clinical Microbiology, April 2008, p. 1220-1225, Vol. 46, No. 4
0095-1137/08/$08.00+0     doi:10.1128/JCM.02158-07
Copyright © 2008, American Society for Microbiology. All Rights Reserved.





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