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Rapid Adaptation of Established High-Throughput Molecular Testing Infrastructure for Monkeypox Virus Detection
Emerging Infectious Diseases
|July 29, 2022
Summary
A new dual-target quantitative PCR assay effectively detects monkeypox virus. This high-throughput method shows excellent analytical and clinical performance, aiding in controlling monkeypox outbreaks.
Area of Science:
- Virology
- Molecular Biology
- Epidemiology
Background:
- The global spread of monkeypox (mpox) in 2022 necessitated rapid diagnostic solutions.
- Existing diagnostic methods required adaptation for high-throughput screening.
Purpose of the Study:
- To develop and validate a dual-target quantitative PCR (qPCR) assay for monkeypox virus detection.
- To adapt the assay for automated, high-throughput platforms commonly used for other viral diagnostics.
Main Methods:
- Adapted two published qPCR assays into a single, dual-target test.
- Determined analytical sensitivity using serial dilutions of monkeypox virus quantified by digital PCR.
- Evaluated clinical performance by comparing results with a commercial orthopoxvirus PCR kit using remnant swab samples.
Main Results:
- The assay demonstrated a lower limit of detection of 4.795 copies/mL.
- Achieved 100% positive agreement (n=11) and 100% negative agreement (n=56) compared to a commercial kit.
- The assay is suitable for high-throughput, automated PCR systems.
Conclusions:
- The developed dual-target qPCR assay provides a scalable and timely diagnostic tool for monkeypox virus.
- Streamlining testing on existing platforms can facilitate efficient public health responses to mpox outbreaks.
