Winstanley Catherine E, Ramsey Kyle H, Marsh Peter, Clarke Ian N
Molecular Microbiology Group, Faculty of Medicine, University of Southampton, Southampton General Hospital, Southampton, Hampshire, United Kingdom.
Department of Microbiology & Immunology, Chicago College of Osteopathic Medicine, Midwestern University, Downers Grove, IL, USA.
J Immunol Methods. 2017 Jun;445:23-30. doi: 10.1016/j.jim.2017.03.002. Epub 2017 Mar 7.
Urogenital infection with Chlamydia trachomatis is the most commonly diagnosed sexually transmitted infection in the developed world. Accurate measurement and therefore understanding the seroprevalence of urogenital C. trachomatis infections requires a rigorously optimised and validated ELISA. Previous ELISAs based on the C. trachomatis plasmid-encoded protein, PGP3, have been described but lack standardisation and critical controls or use a less common PGP3 as the capture antigen.
METHODOLOGY/PRINCIPAL FINDINGS: A sensitive and specific indirect ELISA was developed based on recombinant PGP3 derived from a urogenital strain of C. trachomatis, serovar E (pSW2), using a rigorous validation protocol. Serum samples were collected from 166 genitourinary medicine (GUM) clinic patients diagnosed as positive or negative for urogenital C. trachomatis infection by nucleic acid amplification testing (NAATs). Overall sensitivity and specificity compared to NAATs was 68.18% and 98.0%, respectively. Sensitivities for female and male samples were 71.93% and 64.15%, respectively. Comparison of samples from these patients diagnosed positive for C. trachomatis by NAAT and patients diagnosed negative by NAAT revealed statistical significance (p≤0.0001).
We have developed and validated a sensitive and specific ELISA to detect anti-PGP3 antibodies as an indicator of past and current infection to C. trachomatis using PGP3 from a common urogenital strain. It is anticipated that this assay will be used for seroepidemiological analysis of urogenital C. trachomatis in populations.
沙眼衣原体引起的泌尿生殖系统感染是发达国家最常被诊断出的性传播感染。准确测量并进而了解泌尿生殖系统沙眼衣原体感染的血清流行率需要经过严格优化和验证的酶联免疫吸附测定（ELISA）。先前已有基于沙眼衣原体质粒编码蛋白PGP3的ELISA方法，但缺乏标准化和关键对照，或者使用较不常见的PGP3作为捕获抗原。
方法/主要发现：基于源自泌尿生殖系统沙眼衣原体血清型E（pSW2）菌株的重组PGP3，采用严格的验证方案，开发了一种灵敏且特异的间接ELISA。通过核酸扩增检测（NAATs）从166名被诊断为泌尿生殖系统沙眼衣原体感染阳性或阴性的泌尿生殖医学（GUM）门诊患者中采集血清样本。与NAATs相比，总体灵敏度和特异性分别为68.18%和98.0%。女性和男性样本的灵敏度分别为71.93%和64.15%。对这些通过NAATs诊断为沙眼衣原体阳性的患者和诊断为阴性的患者的样本进行比较，具有统计学意义（p≤0.0001）。
我们开发并验证了一种灵敏且特异的ELISA，以检测抗PGP3抗体，作为过去和当前沙眼衣原体感染的指标，该ELISA使用来自常见泌尿生殖系统菌株的PGP3。预计该检测方法将用于人群中泌尿生殖系统沙眼衣原体的血清流行病学分析。
