Sachdeva Poonam, Patel Achchhe Lal, Sachdev Divya, Ali Mashook, Mittal Aruna, Saluja Daman
Dr B. R. Ambedkar Center for Biomedical Research, University of Delhi, Delhi, India.
Institute of Pathology (ICMR), Safdarjung Hospital Campus, New Delhi, India.
J Med Microbiol. 2009 Jul;58(Pt 7):867-873. doi: 10.1099/jmm.0.008698-0. Epub 2009 Jun 5.
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.
为改善印度沙眼衣原体感染的控制情况，迫切需要一种快速、特异且经济高效的方法。我们通过靶向沙眼衣原体磷脂酶D核酸内切酶超家族中一个编码蛋白质的独特基因组序列，开发了一种内部聚合酶链反应（PCR）检测方法，该方法可产生一个368 bp的扩增片段。使用来自其他性传播疾病致病及相关微生物和人类的基因组DNA，确认了引物的特异性。该检测方法高度灵敏，能够检测低至10 fg的沙眼衣原体DNA。使用450份宫颈标本进行了内部开发的PCR的临床评估，这些标本被分为两组。在第一组（n = 274）中，将内部PCR与直接荧光检测法进行了比较。内部PCR方法的灵敏度为97.22%，而直接荧光抗体检测法为88%。在第二组（n = 176）中，将内部PCR与罗氏AMPLICOR MWP CT检测商业试剂盒进行了比较。本文报道的内部PCR检测方法的灵敏度为93.1%，特异性为97.46%，使其成为沙眼衣原体生殖器感染常规诊断的一种经济高效的替代方法。该方法应有助于早期检测，从而更好地预防和治疗印度的生殖器感染。
