title: A multiplexed microfluidic PCR assay for sensitive and specific point-of-care detection of Chlamydia trachomatis.
abstract: BACKGROUND
  Chlamydia trachomatis (Ct) is the most common cause of bacterial sexually transmitted diseases (STD) worldwide. While commercial nucleic acid amplification tests (NAAT) are available for Ct, none are rapid or inexpensive enough to be used at the point-of-care (POC). Towards the first Ct POC NAAT, we developed a microfluidic assay that simultaneously interrogates nine Ct loci in 20 minutes.
  
  
  RESULTS
  Endocervical samples were selected from 263 women at high risk for Ct STDs (∼35% prevalence). A head-to-head comparison was performed with the Roche-Amplicor NAAT. 129 (49.0%) and 88 (33.5%) samples were positive by multiplex and Amplicor assays, respectively. Sequencing resolved 71 discrepant samples, confirming 53 of 53 positive multiplex samples and 12 of 18 positive Amplicor samples. The sensitivity and specificity were 91.5% and 100%, and 62.4% and 95.9%, respectively, for multiplex and Amplicor assays. Positive and negative predictive values were 100% and 91%, and 94.1% and 68.6%, respectively.
  
  
  CONCLUSIONS
  This is the first rapid multiplex approach to Ct detection, and the assay was also found to be superior to a commercial NAAT. In effect, nine simultaneous reactions significantly increased sensitivity and specificity. Our assay can potentially increase Ct detection in globally diverse clinical settings at the POC.
authors.affiliation: Center for Immunobiology and Vaccine Development, Children's Hospital Oakland Research Institute, Oakland, California, United States of America.
authors.forename: Deborah
authors.initials: D
authors.lastname: Dean
authors.forename: Rosemary S
authors.initials: RS
authors.lastname: Turingan
authors.forename: Hans-Ulrich
authors.initials: HU
authors.lastname: Thomann
authors.forename: Anna
authors.initials: A
authors.lastname: Zolotova
authors.forename: James
authors.initials: J
authors.lastname: Rothschild
authors.forename: Sandeep J
authors.initials: SJ
authors.lastname: Joseph
authors.forename: Timothy D
authors.initials: TD
authors.lastname: Read
authors.forename: Eugene
authors.initials: E
authors.lastname: Tan
authors.forename: Richard F
authors.initials: RF
authors.lastname: Selden
mesh_terms: D000293:Adolescent; D000328:Adult; D002690:Chlamydia Infections / Q000175:diagnosis* / Q000453:epidemiology; D002692:Chlamydia trachomatis / Q000235:genetics / Q000302:isolation & purification*; D005260:Female; D006801:Humans; D044085:Microfluidics; D060885:Multiplex Polymerase Chain Reaction; D019095:Point-of-Care Systems; D015995:Prevalence; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D055815:Young Adult