1887

Journal of Medical Microbiology

Volume 58, Issue 7

Comparison of an in-house PCR assay, direct fluorescence assay and the Roche AMPLICOR kit for detection of Free

Abstract

To improve the control of 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 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 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 (=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 (=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 . The method should facilitate early detection leading to better prevention and treatment of genital infection in India.

  • Received:
  • Accepted:
  • Published Online:
Keyword(s): DFA, direct fluorescence assay , EBs, elementary bodies , FISH, fluorescence in situ hybridization , NAAT, nucleic acid amplification test , NPV, negative predictive value , PPV, positive predictive value and STD, sexually transmitted disease
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2009-07-01
2021-10-26
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Comparison of an in-house PCR assay, direct fluorescence assay and the Roche AMPLICOR Chlamydia trachomatis kit for detection of C. trachomatis
J. Med. Microbiol. 58, 867 (2009); https://doi.org/10.1099/jmm.0.008698-0
/content/journal/jmm/10.1099/jmm.0.008698-0
/content/journal/jmm/10.1099/jmm.0.008698-0
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