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Journal of Clinical Microbiology, March 1999, p. 575-580, Vol. 37, No. 3
0095-1137/99/$00.00+0

Rapid Detection of the Chlamydiaceae and Other Families in the Order Chlamydiales: Three PCR Tests

Karin D. E. Everett,* Linda J. Hornung, and Arthur A. Andersen

Avian and Swine Respiratory Diseases Research Unit, USDA Agricultural Research Service, National Animal Disease Center, Ames, Iowa 50010

Received 6 October 1998/Accepted 8 December 1998

Few identification methods will rapidly or specifically detect all bacteria in the order Chlamydiales, family Chlamydiaceae. In this study, three PCR tests based on sequence data from over 48 chlamydial strains were developed for identification of these bacteria. Two tests exclusively recognized the Chlamydiaceae: a multiplex test targeting the ompA gene and the rRNA intergenic spacer and a TaqMan test targeting the 23S ribosomal DNA. The multiplex test was able to detect as few as 200 inclusion-forming units (IFU), while the TaqMan test could detect 2 IFU. The amplicons produced in these tests ranged from 132 to 320 bp in length. The third test, targeting the 23S rRNA gene, produced a 600-bp amplicon from strains belonging to several families in the order Chlamydiales. Direct sequence analysis of this amplicon has facilitated the identification of new chlamydial strains. These three tests permit ready identification of chlamydiae for diagnostic and epidemiologic study. The specificity of these tests indicates that they might also be used to identify chlamydiae without culture or isolation.

* Corresponding author. Present address: Department of Medical Microbiology/Parasitology, College of Veterinary Medicine, University of Georgia, Athens, GA 30602-7371. Phone: (706) 542-5823. Fax: (706) 542-5771. E-mail: keverett{at}calc.vet.uga.edu or kdeeverett{at}hotmail.com.

Journal of Clinical Microbiology, March 1999, p. 575-580, Vol. 37, No. 3
0095-1137/99/$00.00+0


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