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Journal of Clinical Microbiology, January 2009, p. 175-181, Vol. 47, No. 1
0095-1137/09/$08.00+0     doi:10.1128/JCM.01851-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.

Genotyping of Chlamydophila psittaci by Real-Time PCR and High-Resolution Melt Analysis{triangledown}

Stephanie L. Mitchell,1 Bernard J. Wolff,1 W. Lanier Thacker,1 Paula G. Ciembor,2 Christopher R. Gregory,2 Karin D. E. Everett,3 Branson W. Ritchie,2 and Jonas M. Winchell1*

Respiratory Disease Branch, Centers for Disease Control and Prevention, Atlanta, Georgia,1 Infectious Diseases Laboratory, College of Veterinary Medicine, University of Georgia, Athens, Georgia,2 Ameritek USA, Everett, Washington3

Received 24 September 2008/ Returned for modification 29 October 2008/ Accepted 31 October 2008

Human infection with Chlamydophila (Chlamydia) psittaci can lead to psittacosis, a disease that occasionally results in severe pneumonia and other medical complications. C. psittaci is currently grouped into seven avian genotypes: A through F and E/B. Serological testing, outer membrane protein A (ompA) gene sequencing, and restriction fragment length polymorphism analysis are currently used for distinguishing these genotypes. Although accurate, these methods are time-consuming and require multiple confirmatory tests. By targeting the ompA gene, a real-time PCR assay has been developed to rapidly detect and genotype C. psittaci by light-upon-extension chemistry and high-resolution melt analysis. Using this assay, we screened 169 animal specimens; 98 were positive for C. psittaci (71.4% genotype A, 3.1% genotype B, 4.1% genotype E, and 21.4% unable to be typed). This test may provide insight into the distribution of each genotype among specific hosts and provide epidemiological and epizootiological data in human and mammalian/avian cases. This diagnostic assay may also have veterinary applications during chlamydial outbreaks, particularly with respect to identifying the sources and tracking the movements of a particular genotype when multiple animal facilities are affected.

* Corresponding author. Mailing address: Centers for Disease Control and Prevention, 1600 Clifton Rd. NE, MS G-03, Atlanta, GA 30333. Phone: (404) 639-4921. Fax: (404) 639-4215. E-mail: jwinchell{at}cdc.gov

{triangledown} Published ahead of print on 12 November 2008.

Journal of Clinical Microbiology, January 2009, p. 175-181, Vol. 47, No. 1
0095-1137/09/$08.00+0     doi:10.1128/JCM.01851-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.





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