DDL Diagnostic Laboratory, Voorburg, The Netherlands.
J Mol Diagn. 2011 Mar;13(2):152-9. doi: 10.1016/j.jmoldx.2010.11.017.
A novel Chlamydia trachomatis (Ct) microsphere suspension (MS) assay was evaluated for identification of the different serovars, using the same PCR primer set established for the Ct Detection and genoTyping assay. Both assays can detect and identify all 14 major serovars (A, B/Ba, C, D/Da, E, F, G/Ga, H, I/Ia, J, K, L1, L2/L2a, and L3) and one genovariant of serovar J. The probe specificity for the Ct-MS assay was determined using 14 Ct reference strains and 1 clinical isolate from a genovariant of serovar J. Also, the Ct-MS assay and the Ct detection and genoTyping assay were compared in 712 Ct-positive clinical samples. The Ct-MS assay showed a highly specific reaction for all probes with the amplicons of the reference strains, giving a very low background median fluorescence intensity signal (median fluorescence intensity ≤ 10). An excellent overall agreement in the Ct detection (kappa = 0.947, 95% confidence interval, 0.89 to 0.999; McNemar's test, P = 1.000) and the Ct genotyping (kappa = 0.993, 95% confidence interval, 0.977 to 1.000; McNemar's test, P = 0.053) was observed between the Ct detection and genoTyping (DT) assay and the Ct-MS assay. In conclusion, the novel Ct-MS assay permits simultaneous detection and genotyping of Ct serovars, making the Ct-MS assay an excellent high throughput method.
一种新型沙眼衣原体（Ct）微球悬浮液（MS）检测方法，使用相同的 Ct 检测和基因分型检测中建立的 PCR 引物组，用于鉴定不同的血清型。这两种检测方法均可检测和鉴定所有 14 种主要血清型（A、B/Ba、C、D/Da、E、F、G/Ga、H、I/Ia、J、K、L1、L2/L2a 和 L3）和 1 种 J 血清型的基因变异体。使用 14 株 Ct 参考株和 1 株 J 血清型基因变异体的临床分离株，确定 Ct-MS 检测的探针特异性。此外，在 712 份 Ct 阳性临床样本中比较了 Ct-MS 检测和 Ct 检测和基因分型检测。Ct-MS 检测对所有探针均具有高度特异性，与参考株的扩增子反应，背景中位荧光强度信号非常低（中位荧光强度≤10）。Ct 检测（kappa = 0.947，95%置信区间，0.89 至 0.999；McNemar 检验，P = 1.000）和 Ct 基因分型（kappa = 0.993，95%置信区间，0.977 至 1.000；McNemar 检验，P = 0.053）的结果在 Ct 检测和基因分型（DT）检测和 Ct-MS 检测之间具有极好的一致性。综上所述，新型 Ct-MS 检测方法可同时检测和鉴定 Ct 血清型，是一种出色的高通量方法。
