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J Med Microbiol 58 (2009), 867-873; DOI: 10.1099/jmm.0.008698-0
© 2009 Society for General Microbiology
ISSN 0022-2615

Comparison of an in-house PCR assay, direct fluorescence assay and the Roche AMPLICOR Chlamydia trachomatis kit for detection of C. trachomatis

Poonam Sachdeva1, Achchhe Lal Patel1, Divya Sachdev1, Mashook Ali1, Aruna Mittal2 and Daman Saluja1

1 Dr B. R. Ambedkar Center for Biomedical Research, University of Delhi, Delhi, India

2 Institute of Pathology (ICMR), Safdarjung Hospital Campus, New Delhi, India

Correspondence
Daman Saluja
dsalujach{at}yahoo.com

Received December 11, 2008
Accepted March 9, 2009

To improve the control of Chlamydia trachomatis infection in India, a rapid, specific and cost-effective method is much needed. We developed an in-house PCR assay by targeting a unique genomic sequence encoding a protein from the C. trachomatis phospholipase D endonuclease superfamily that produces an amplified fragment of 368 bp. The specificity of the primers was confirmed using genomic DNA from other sexually transmitted disease-causing and related micro-organisms and from humans. The assay was highly sensitive and could detect as low as 10 fg C. trachomatis DNA. Clinical evaluation of the in-house-developed PCR was carried out using 450 endocervical specimens that were divided in two groups. In group I (n=274), in-house PCR was evaluated against the direct fluorescence assay. The resolved sensitivity of the in-house PCR method was 97.22 % compared with 88 % for the direct fluorescent antibody assay. In group II (n=176), the in-house PCR was compared with the commercial Roche AMPLICOR MWP CT detection kit. The resolved sensitivity of the in-house PCR assay reported here was 93.1 % and the specificity was 97.46 %, making it a cost-effective alternative for routine diagnosis of genital infection by C. trachomatis. The method should facilitate early detection leading to better prevention and treatment of genital infection in India.

Supplementary figures are available with the online version of this paper.






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