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Journal of Clinical Microbiology, December 1998, p. 3567-3573, Vol. 36, No. 12
0095-1137/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.

Differentiation of Campylobacter jejuni Serotype O19 Strains from Non-O19 Strains by PCR

Naoaki Misawa,1 Ban Mishu Allos,1 and Martin J. Blaser1,2,*

Division of Infectious Diseases, Vanderbilt University School of Medicine, Nashville, Tennessee 37232-2605,1 and Department of Veterans Affairs Medical Center, Nashville, Tennessee 372122

Received 7 May 1998/Returned for modification 17 August 1998/Accepted 23 September 1998

Guillain-Barré syndrome (GBS), a neurologic disease characterized by acute paralysis, is frequently preceded by Campylobacter jejuni infection. Serotype O19 strains are overrepresented among GBS-associated C. jejuni isolates. We previously showed that all O19 strains tested were closely related to one another by randomly amplified polymorphic DNA (RAPD) and restriction fragment length polymorphism analyses. RAPD analysis demonstrated a 1.4-kb band in all O19 strains tested but in no non-O19 strains. We cloned this O19-specific band; nucleotide sequence analysis revealed a truncated open reading frame with significant homology to DNA gyrase subunit B (gyrB) of Helicobacter pylori. PCR using the random primer and a primer specific for gyrB showed that in non-O19 strains, the random primer did not recognize the downstream gyrB binding site. The regions flanking each of the random primer binding sites were amplified by degenerate PCR for further sequencing. Although the random primer had several mismatches with the downstream gyrB binding site, a single nucleotide polymorphism 6 bp upstream from the 3' terminus was found to distinguish O19 and non-O19 strains. PCR using 3'-mismatched primers based on this polymorphism was designed to differentiate O19 strains from non-O19 strains. When a total of 42 (18 O19 and 24 non-O19) strains from five different countries were examined, O19 strains were distinguishable from non-O19 strains in each case. This PCR method should permit identification of O19 C. jejuni strains.

* Corresponding author. Mailing address: Division of Infectious Diseases, Vanderbilt University School of Medicine, A-3310 Medical Center North, Nashville, TN 37232-2605. Phone: (615) 322-2035. Fax: (615) 343-6160. E-mail: Martin.Blaser{at}mcmail.vanderbilt.edu.

Journal of Clinical Microbiology, December 1998, p. 3567-3573, Vol. 36, No. 12
0095-1137/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.


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