1887

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Volume 66, Issue 3

Development, validation and testing costs of an in-house real-time PCR assay for the detection of Free

Abstract

To improve the screening of 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 infection.

Asymptomatic women aged 14–25 years who attended primary health services in Manaus, Brazil, were screened for 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, 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.

The primers were considered specific for 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 was £0.44 ($0.55) for HCII CT-ID, £1.16 ($1.45) for Artus qPCR and £1.06 ($1.33) for in-house qPCR.

We have standardized an in-house qPCR to detect cervical targeting CP. The in-house qPCR showed excellent accuracy and was more affordable than the commercial qPCR kit.

  • Received:
  • Accepted:
  • Published Online:
Keyword(s): Amazonas , Chlamydia trachomatis , cost , real-time polymerase chain reaction and validation studies
© 2017 The Authors
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2017-03-01
2024-04-24
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Development, validation and testing costs of an in-house real-time PCR assay for the detection of Chlamydia trachomatis
J. Med. Microbiol. 66, 312 (2017); https://doi.org/10.1099/jmm.0.000443
/content/journal/jmm/10.1099/jmm.0.000443
/content/journal/jmm/10.1099/jmm.0.000443
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