Performance of a Novel Real-Time PCR-Based Assay for Rapid Monkeypox Virus Detection in Human Samples

Abstract

The ongoing epidemic of mpox, namely human monkeypox virus (MPXV) infection, requires rapid and reliable laboratory diagnosis. We report on the QIAstat-Dx viral vesicular panel PCR assay that allows the detection of (within 75 min) six vesicular disease-causing viruses, including MPXV. We analyzed 168 clinical samples, known to be positive (51 samples) or negative (117 samples) for MPXV clade II, obtained from patients at their mpox diagnosis or follow-up. QIAstat assay results were compared to those of a MPXV-specific reference PCR assay. The QIAstat assay detected MPXV (clade II) in 51 (100%) of 51 samples and did not detect MPXV in 117 (100%) of 117 samples, resulting in a positive or negative agreement of 100% (95% CI, 93.0–100) and 100% (95% CI, 96.8–100), respectively. Of the 20 patients diagnosed with mpox, 18 (90.0%) had at least a vesicular swab and 1 (5.0%) had only an oropharyngeal swab positive for MPXV. At mpox follow-ups, 2 (10.0%) of 20 patients had first-time positive whole blood samples. Thirteen MPXV-negative samples were positive for mpox-mimicking viruses. Our findings show the excellent performance of the QIAstat-Dx assay for MPXV detection in clinical samples. Further studies are needed before considering a large-scale application of the QIAstat-Dx assay.

A field diagnostic method for rapid and sensitive detection of mpox virus

“…25 In addition to antigen/antibody-based serology tests, PCR-based diagnosis has also becoming widely adopted as a result of its specificity and high sensitivity. 31,32 These assays often need laboratory equipment and involve complex procedures, which discourage their utility as pointof-care tools particularly in resource-limited settings. In this context, isothermal amplification technologies present advantage in their independence on special laboratory instruments and their feasibility of being developed into molecular diagnostic tools for field testing.…”

“…Rapid and accurate diagnostic methods are the foundation of effective prevention and control of mpox outbreaks 25 . In addition to antigen/antibody‐based serology tests, PCR‐based diagnosis has also becoming widely adopted as a result of its specificity and high sensitivity 31,32 . These assays often need laboratory equipment and involve complex procedures, which discourage their utility as point‐of‐care tools particularly in resource‐limited settings.…”