Development, validation and testing costs of an in-house real-time PCR assay for the detection of Chlamydia trachomatis
Development, validation and testing costs of an in-house real-time PCR assay for the detection of Chlamydia trachomatis
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DOI：
10.1099/jmm.0.000443
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发表时间：
2017-03-01
影响因子：
3
通讯作者：
Benzaken, Adele Schwartz
中科院分区：
文献类型：
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作者：
dos Santos, Camila Gurgel;Sabido, Meritxell;Benzaken, Adele Schwartz
目的.提高沙眼衣原体（C。在巴西，需要一种准确和负担得起的方法。本研究的目的是开发和评估一种新的内部实时PCR（qPCR）检测诊断C。沙眼感染.在巴西马瑙斯的初级卫生服务机构就诊的14-25岁无症状妇女接受了C.使用Digene Hybrid Capture II CT-ID（HCII CT-ID）DNA测试检测沙眼衣原体。使用内部qPCR和市售qPCR（Artus C）检测宫颈标本子集。trachomatis Plus RG PCR 96 CE（Artus qPCR）试剂盒，作为参考检测。根据隐蔽质粒（CP）序列设计引物/探针。从供应商的角度进行了经济评价。认为引物对C.沙眼衣原体，因为他们没有扩增任何产品从非性传播的细菌物种测试。总体而言，通过市售试剂盒（Artus qPCR）和内部qPCR检测了292份标本。其中，1例未导致扩增，从分析中排除。内部qPCR的灵敏度、特异性以及阳性和阴性预测值分别为99.5% [95%置信区间（CI）：97.1-100]、95.1%（95% CI：89-98.4）、97.4%（95% CI：94-99.1）和99.0%（95% CI：94.5-100）。每例C. HCII CT-ID检测沙眼衣原体的平均阳性率为0.44磅（$0.55），Artus qPCR检测沙眼衣原体的平均阳性率为1.16磅（$1.45），内部qPCR检测沙眼衣原体的平均阳性率为1.06磅（$1.33）。我们已将内部qPCR标准化，以检测宫颈C。沙眼衣原体靶向CP。内部qPCR显示出极好的准确性，并且比商业qPCR试剂盒更经济实惠。
Purpose. To improve the screening of Chlamydia trachomatis (C. trachomatis) in Brazil, an accurate and affordable method is needed. The objective of this study was to develop and assess the performance and costs of a new in-house real-time PCR (qPCR) assay for the diagnosis of C. trachomatis infection.Methodology. Asymptomatic women aged 14-25 years who attended primary health services in Manaus, Brazil, were screened for C. trachomatis using the Digene Hybrid Capture II CT-ID (HCII CT-ID) DNA test. A subset of cervical specimens were tested using an in-house qPCR and a commercial qPCR, Artus C. trachomatis Plus RG PCR 96 CE (Artus qPCR) kit, as a reference test. A primer/probe based on the sequence of cryptic plasmid (CP) was designed. An economic evaluation was conducted from the provider's perspective.Results. The primers were considered specific for C. trachomatis because they did not amplify any product from non-sexually transmitted bacterial species tested. Overall, 292 specimens were tested by both the commercial kit (Artus qPCR) and the in-house qPCR. Of those, one resulted in no amplification and was excluded from the analysis. The sensitivity, specificity, and positive and negative predictive values of the in-house qPCR were 99.5% [95% confidence interval (CI): 97.1-100], 95.1% (95% CI: 89-98.4), 97.4% (95% CI: 94-99.1) and 99.0% (95% CI: 94.5-100), respectively. The cost per case of C. trachomatis was 0.44 pound ($0.55) for HCII CT-ID, 1.16 pound ($1.45) for Artus qPCR and 1.06 pound ($1.33) for in-house qPCR.Conclusion. We have standardized an in-house qPCR to detect cervical C. trachomatis targeting CP. The in-house qPCR showed excellent accuracy and was more affordable than the commercial qPCR kit.