Comparison of an in-house PCR assay, direct fluorescence assay and the Roche AMPLICOR Chlamydia trachomatis kit for detection of C. trachomatis.
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.
Sachdeva
Poonam
P
Dr B. R. Ambedkar Center for Biomedical Research, University of Delhi, Delhi, India.
Patel
Achchhe Lal
AL
Dr B. R. Ambedkar Center for Biomedical Research, University of Delhi, Delhi, India.
Sachdev
Divya
D
Dr B. R. Ambedkar Center for Biomedical Research, University of Delhi, Delhi, India.
Ali
Mashook
M
Dr B. R. Ambedkar Center for Biomedical Research, University of Delhi, Delhi, India.
Mittal
Aruna
A
Institute of Pathology (ICMR), Safdarjung Hospital Campus, New Delhi, India.
Saluja
Daman
D
Dr B. R. Ambedkar Center for Biomedical Research, University of Delhi, Delhi, India.
D000328:Adult; D002690:Chlamydia Infections / Q000175:diagnosis*; D002692:Chlamydia trachomatis / Q000302:isolation & purification*; D017931:DNA Primers / Q000592:standards; D005260:Female; D019085:Fluorescent Antibody Technique, Direct / Q000379:methods*; D006801:Humans; D016133:Polymerase Chain Reaction / Q000379:methods*; D011933:Reagent Kits, Diagnostic; D012680:Sensitivity and Specificity; D013997:Time Factors
D017931:DNA Primers; D011933:Reagent Kits, Diagnostic