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Journal of Clinical Microbiology, January 2003, p. 197-204, Vol. 41, No. 1
0095-1137/03/$08.00+0     DOI: 10.1128/JCM.41.1.197-204.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Genetic Diversity of Neisseria gonorrhoeae Housekeeping Genes

Raphael P. Viscidi* and James C. Demma

Stanley Division of Developmental Neurovirology, Department of Pediatrics, Johns Hopkins University School of Medicine, Baltimore, Maryland 21287

Received 10 June 2002/ Returned for modification 26 July 2002/ Accepted 16 October 2002

Molecular typing of Neisseria gonorrhoeae strains is an important tool for epidemiological studies of gonococcal infection and transmission. The recently developed multilocus sequence typing (MLST) method is based on the genetic variation among housekeeping genes. As a preliminary investigation for the development of such a method, we characterized the genetic diversity at 18 gonococcal housekeeping gene loci. Approximately 17,500 nucleotides, spanning 18 loci, were sequenced from 24 isolates. Including strain FA 1090, which has been fully sequenced, and three unique glnA sequences from GenBank, the number of alleles identified for the 18 loci ranged from 2 to 18, with a mean of 8.3 alleles per locus. The majority of polymorphic sites were distributed randomly along the genes, consistent with evolution of DNA sequences by point mutation. In addition, several examples of clustered mutations and insertions or deletions were detected, which most likely occurred by recombinational events. While purifying selection is the dominant force driving the evolution of these housekeeping genes, positive selection also appeared to operate on the abcZ and gpdh loci. The 25 completely characterized strains each had a unique allelic profile with as few as three loci (pilA, abcZ, and pip or pgi2). Molecular typing based on the allelic profile of housekeeping genes resolved the isolates better than either porB nucleotide sequencing or typing of the opa gene. The allelic profiles for the pilA, abcZ, and serC loci of paired strains from 16 sexual contacts were identical. A potential MLST for N. gonorrhoeae, based on ~500- to 600-bp gene fragments of seven housekeeping gene loci, would include the pilA, abcZ, serC, glnA, gdh, gnd, and pip loci.

* Corresponding author. Mailing address: The Johns Hopkins Hospital, Blalock Building, Room 1105, 600 North Wolfe St., Baltimore, MD 21287. Phone: (410) 614-1494. Fax: (410) 955-3723. E-mail: rviscid1{at}jhem.jhmi.edu.

Journal of Clinical Microbiology, January 2003, p. 197-204, Vol. 41, No. 1
0095-1137/03/$08.00+0     DOI: 10.1128/JCM.41.1.197-204.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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