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P5.061 Microwave-Accelerated Metal-Enhanced Fluorescence (MAMEF) Point-Of-Care Test For the Detection of Chlamydia Trachomatis
  1. J H Melendez1,
  2. J Huppert2,
  3. E Hesse2,
  4. M Jett-Goheen3,
  5. N Quinn3,
  6. C A Gaydos3,
  7. C D Geddes1
  1. 1Institute of Fluorescence UMBC, Baltimore, MD, United States
  2. 2Cincinnati Children’s Hospital Medical Center, Cincinnati, OH, United States
  3. 3Johns Hopkins University School of Medicine, Baltimore, MD, United States

Abstract

Background Chlamydia trachomatis (CT) is the most prevalent bacterial sexually transmitted infection (STI) reported worldwide. Accurate point-of-care diagnostic tests are urgently needed for the rapid treatment of patients. To address this need, we have previously developed a 16S rRNA-based Microwave-Accelerated Metal-Enhanced Fluorescence (MAMEF) assay for the detection of CT. Here we report the development of an additional CT cryptic plasmid-based MAMEF assay, the use of the assays on clinical samples and the implication of MAMEF as a point-of-care test.

Methods The cryptic plasmid-based assay was investigated with cultured, titered CT and vaginal specimens. Following the optimization of the assay, we tested a blinded cohort of dry-shipped vaginal swabs using both the 16S rRNA- and cryptic plasmid-based MAMEF assays, and compared the results to nucleic acid amplification tests (NAATs).

Results The MAMEF assays detected as few as 10 IFU/mL of CT in less than 10 minutes including DNA extraction and detection. A total of 257 vaginal swabs from 245 adolescent women (ages 14–22) were analysed by MAMEF. The overall prevalence of CT by NAAT was 17.5%. Of the 45 NAAT CT-positive samples and 212 CT-negative samples, 33/45 and 197/212 were correctly identified by both MAMEF assays (sens 73.3%, spec 92.9%). Using the plasmid-based assay alone, 37/45 CT+ and 197/212 CT- were detected (sens 82.2%; spec 92.9%). Using the 16S rRNA assay alone, 34/45 CT+ and 197/212 CT- (sens 75.5%; spec 92.9). For the overall % agreement with NAAT, the individual 16S rRNA and cryptic plasmid were 89.8% and 91%, respectively.

Conclusions Both CT MAMEF assays demonstrated moderate sensitivity and high specificity when using dry-shipped vaginal swabs. The cryptic plasmid assay was more sensitive than the 16S rRNA assay. MAMEF is an ultra-rapid platform with results in less than 10 minutes making it ideal as a point-of-care test.

  • chlamydia
  • diagnostics
  • NAAT

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