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This research aims to assess the limit of detection and inclusivity of a prototype diagnostic panel for STIs.

January 14, 2026Open Forum Infectious Diseases0 citationsOpen Access

P-729. Limit of Detection and Inclusivity of a Research-Use-Only (RUO) Prototype of a Multiplexed Sample-to-Answer Diagnostic System for Identification of Sexually Transmitted Infections from Vaginal Swabs and Penile Urine

KKKerrin KochbioMérieux (United States)MMMarta MangifestabioMérieux (United States)CCCaitlin CastanedabioMérieux (United States)

Key Points

This research aims to assess the limit of detection and inclusivity of a prototype diagnostic panel for STIs.
Evaluated limit of detection for various pathogens in RUO prototype.
Conducted serial dilutions of reference isolates to determine genome copies using droplet digital PCR assays.
Tested inclusivity with 43 PCR-quantified strains and performed in silico analysis on available sequences.
Limit of detection varied: 275 GE/mL for N. gonorrhoeae to 0.28 GE/mL for T. vaginalis.
Estimated > 99.7% reactivity for 3 out of 4 analytes based on homology assessment.
T. vaginalis showed 10-fold greater sensitivity in urine samples compared to vaginal swabs.

Abstract

Abstract Background Untreated sexually transmitted infections adversely impact sexual health and reproductive outcomes. bioMérieux is developing the BIOFIRE® SPOTFIRE® Sexually Transmitted Infection (STI) Panel, a molecular test designed for a point-of-care setting to detect Chlamydia trachomatis, Mycoplasma genitalium, Neisseria gonorrhoeae, and Trichomonas vaginalis from vaginal swabs (Vswabs) or penile urine collected in eNAT® medium. This study evaluated the limit of detection (LoD) and inclusivity of the pathogen assays in a RUO panel prototype.Table 1.Limit of Detection and Inclusivity Assessment of the RUO BIOFIRE® SPOTFIRE® Sexually Transmitted Infection Panel Methods Serial dilutions of reference isolates, quantified to determine genome copies using droplet digital PCR assays targeting single-copy genes, were spiked into negative clinical matrices and used to determine the 95% LoD for each pathogen. Inclusivity was assessed empirically using 43 PCR-quantified strains spiked into negative matrices at near-LoD genome equivalent (GE) levels as well as in silico using sequence data available at the time of analysis. Results See Table 1 for study results. The LoD of the reference isolates varied from 275 GE/mL for N. gonorrhoeae to 0.28 GE/mL for T. vaginalis. All analytes besides T. vaginalis had equivalent LoDs in both sample matrices, with 10-fold greater sensitivity for T. vaginalis in contrived urine samples. Inclusivity testing of 43 organism strains at 5-20X LoD showed no reactivity limitations with equivalent reactivity in both sample matrices. In silico assessments based on assay primer homology to 900 available sequences predict 99.7% reactivity for 3 /4 analytes on the panel; an assessment of the degree of reactivity impairment for 2.6% of M. genitalium sequences is planned. Conclusion This study suggests the BIOFIRE® SPOTFIRE® STI Panel will be capable of detecting low levels and multiple strains of clinically relevant C. trachomatis, M. genitalium, N. gonorrhoeae, and T. vaginalis strains from appropriate specimens. Disclosures All Authors: No reported disclosures

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