Liverpool Specialist Virology Centre, Royal Liverpool University Hospital, Liverpool, UK.
Sex Transm Infect. 2010 Jun;86(3):207-11. doi: 10.1136/sti.2009.040634.
To evaluate a sensitive and specific, real-time PCR assay with internal control for Chlamydia trachomatis and Neisseria gonorrhoeae DNA detection in urine specimens.
The diagnostic performance of a laboratory-developed quadruplex assay (LDQA) targeting the cryptic plasmid and MOMP genes of C trachomatis, the porA pseudogene of N gonorrhoeae and a synthetic internal control was assessed using 1028 urine specimens. The LDQA was compared with the Roche COBAS Taqman CT test and the COBAS Amplicor NG assay with supplemental confirmation tests. The subsequent performance of the LDQA in detecting N gonorrhoeae was monitored in comparison with bacterial culture from swabs.
88 (8.6%) urines were determined as C trachomatis positive in the diagnostic evaluation. LDQA sensitivity and specificity were calculated to be 100% and 99.9%, respectively, for C trachomatis. The LDQA showed high specificity with isolates of other Neisseria species and gave complete concordance with resolved data for N gonorrhoeae detection. However, the incidence of N gonorrhoeae infection was low, with 17 (1.7%) positive patients. A post-implementation audit of 14 316 patients gave the LDQA N gonorrhoeae urine PCR protocol (porA, OPA, 16s rDNA) a sensitivity of 96.9% and specificity of 99.8% in comparison with bacterial culture from swabs.
The LDQA was found to be an effective method for the detection of C trachomatis and N gonorrhoeae DNA in urine samples, and the PCR protocol has replaced bacterial culture for the screening of N gonorrhoeae in asymptomatic men and women in the laboratory.
评估一种灵敏且特异的实时 PCR 检测方法，该方法采用内对照检测尿液标本中的沙眼衣原体和淋病奈瑟菌 DNA。
采用实验室自建的四重检测方法（LDQA），检测沙眼衣原体隐秘质粒和 MOMP 基因、淋病奈瑟菌 porA 假基因和合成内对照，评估其对 1028 例尿液标本的诊断性能。将 LDQA 与罗氏 COBAS Taqman CT 检测和 COBAS Amplicor NG 检测进行比较，并与拭子细菌培养进行补充确认检测。随后，与拭子细菌培养相比，监测 LDQA 检测淋病奈瑟菌的后续性能。
在诊断评估中，88（8.6%）例尿液被确定为沙眼衣原体阳性。LDQA 的敏感性和特异性分别为 100%和 99.9%，用于检测沙眼衣原体。LDQA 对其他奈瑟菌种的分离株具有高特异性，并与淋病奈瑟菌检测的解析数据完全一致。然而，淋病奈瑟菌感染的发生率较低，仅 17（1.7%）例阳性患者。对 14316 例患者进行实施后审核，LDQA 淋病奈瑟菌尿液 PCR 检测方案（porA、OPA、16s rDNA）与拭子细菌培养相比，敏感性为 96.9%，特异性为 99.8%。
LDQA 被发现是一种有效的检测方法，可用于检测尿液样本中的沙眼衣原体和淋病奈瑟菌 DNA，并且该 PCR 方案已取代细菌培养，用于实验室中无症状男性和女性淋病奈瑟菌的筛查。
