Development, validation and testing costs of an in-house real-time polymerase chain reaction assay for the detection of Chlamydia trachomatis.

The objective of this study was to develop and assess the performance and costs of a new in-house real time polymerase chain reaction (qPCR) assay for the diagnosis of CT infection. Asymptomatic women aged 14-25 years who attended primary health services in Manaus, Brazil, were screened for CT using the Digene® Hybrid Capture II (HCII CT-ID) CT-ID DNA-test. A subset of cervical specimens were tested using an in-house qPCR and a commercial qPCR (Artus qPCR) as a reference test. A primer/probe based on the sequence of cryptic plasmid was designed. An economic evaluation was conducted from the provider's perspective. The primers were considered specific for CT because they did not amplify any product from non-STI tested species. Overall, 292 specimens were tested by both the commercial kit (qPCR Artus Plus CT) and the in-house qPCR. Of those, one resulted in no amplification and was excluded from the analysis. The sensitivity, specificity, positive and negative predictive values of in 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 CT was $0.55 for HCII CT-ID, $1.45 for Artus qPCR and $1.33 for in-house qPCR. We have standardized an in-house qPCR to detect cervical CT targeting cryptic plasmid. The in-house qPCR showed excellent accuracy and was more affordable than the qPCR commercial kit. PMID: 28141509 [PubMed - as supplied by publ...
Source: Journal of Medical Microbiology - Category: Microbiology Authors: Tags: J Med Microbiol Source Type: research