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Journal of Clinical Microbiology, September 2001, p. 3072-3079, Vol. 39, No. 9
0095-1137/01/$04.00+0   DOI: 10.1128/JCM.39.9.3072-3079.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

PCR Detection and Molecular Identification of Chlamydiaceae Species

J. C. Hartley,1,* S. Kaye,2 S. Stevenson,1 J. Bennett,2 and G. Ridgway1

Department of Clinical Microbiology, University College London Hospital NHS Trust, London WC1E 6DB,1 and Royal Free and University College Medical School, Department of Virology, London W1T 4JF,2 United Kingdom

Received 16 November 2000/Returned for modification 18 December 2000/Accepted 21 May 2001

Recent taxonomic developments, based on 16s and 23s rRNA gene sequences, have divided the family Chlamydiaceae into two genera and nine species, of which five have been found to infect humans. Few simple methods are available to detect and identify all species sensitively and specifically. In this study the suitability of the omp2 gene as a target for molecular identification of Chlamydiaceae is demonstrated. Phylogenetic analysis of partial omp2 gene sequences from all nine species agrees with the recently published taxonomic changes based on the ribosomal genes. The use of a family-specific PCR primer pair, which is able to amplify the 5' end of the omp2 gene from all Chlamydiaceae except some Chlamydophila pecorum strains, is described. Identification of all nine species was achieved using restriction fragment length polymorphism analysis with a single enzyme, AluI, confirmed by DNA sequencing. A PCR enzyme-linked oligonucleotide assay was developed which can detect a single chlamydial genome and may be applied to DNA extracts from any specimen or culture for the detection of single or mixed human chlamydial infection.

* Corresponding author. Present address: Department of Microbiology, Level 4, Camelia Botnar Laboratories, Great Ormond Street Hospital for Children NHS Trust, Great Ormond St., London WC1N 3JH, United Kingdom. Phone: (0)20 7405 9200, ext. 5285. Fax: (0)20 7813 8268. E-mail: HartlJ3{at}gosh.nhs.uk.

Journal of Clinical Microbiology, September 2001, p. 3072-3079, Vol. 39, No. 9
0095-1137/01/$04.00+0   DOI: 10.1128/JCM.39.9.3072-3079.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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