ArticleJournal of clinical virology : the official publication of the Pan American Society for Clinical Virology2023
Evaluation and clinical validation of monkeypox (mpox) virus real-time PCR assays.
Article in Journal of clinical virology : the official publication of the Pan American Society for Clinical Virology, 2023. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 28 papers.
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Who cites it
28 citing papers in PubMed, 57 citations in OpenAlex.
- Tecovirimat treatment of clade II mpox: a virological analysis of a randomised, double-blind, phase 3 trial.The Lancet. Microbe · 2026Trial
- One-Pot RPA-CRISPR/Cas12a Assay With Visual Readout for the Ultra-Specific Detection of Monkeypox Virus Clade I.Microbial biotechnology · 2026Article
- Comparison of diagnostic performance between the World Health Organization and the Centers for Disease Control and Prevention real-time PCR assays for molecular detection of the mpox virus.Microbiology spectrum · 2026Article
- Severe Clade I (subclade Ib) mpox in advanced HIV complicated by multidrug-resistant bacterial superinfection and ocular involvement: a case report from Uganda.Tropical medicine and health · 2026Article
- Evaluation and comparison of three qPCR commercial assays and three automated platforms for the detection of monkeypox virus DNA.Microbiology spectrum · 2026Article
- Saliva, urine, and anorectal swabs as viable alternatives to lesion swabs for molecular diagnosis of mpox.Nature communications · 2026Article
- Evaluation and Clinical Validation of Pan-Specific and Clade-Specific Diagnostic Real-Time PCR Assays for Monkeypox Virus.Journal of medical virology · 2026Article
- Differentiating mpox infection and vaccination using a validated multiplex orthopoxvirus IgG serology assay.Journal of clinical microbiology · 2026Article
- Real-time public health interventions driven by wastewater and environmental surveillance during Uganda's 2025 martyrs day mass gathering.Journal of health, population, and nutrition · 2026Article
- Research Trends and Development Dynamics of qPCR-based Biomarkers: A Comprehensive Bibliometric Analysis.Molecular biotechnology · 2026Review
- Development and evaluation of a two-tube multiplex real-time PCR assay for differential detection of Monkeypox virus clades Ia, Ib, IIa, and IIb.Frontiers in microbiology · 2026Article
- LAMPOX: A Portable and Rapid Molecular Diagnostic Assay for the Epidemic Clade IIb Mpox Virus Detection.Diagnostics (Basel, Switzerland) · 2025Article
- Development and Evaluation of a Molecular Test for Monkeypox Virus in the Federal District, Brazil.Genes · 2025Article
- Complete genome of an mpox clade 1b virus from Kenya.Microbiology resource announcements · 2025Article
- Mpox disease, diagnosis, and point of care platforms.Bioengineering & translational medicine · 2025Review
- Clade I or clade II? Targeting essential viral genes to differentiate monkeypox virus clades by multiplex real-time PCR.Frontiers in public health · 2025Article
- Development of a two-tube multiplex real-time fluorescent PCR for the simultaneous differentiation of the mpox virus clades and the A.1, B.1 and C.1 lineages within clade IIb.Frontiers in cellular and infection microbiology · 2025Article
- Monkeypox (Mpox): Diagnosis and Emerging Challenges.The Yale journal of biology and medicine · 2024Review
- Advances and challenges of mpox detection technology.Biosafety and health · 2024Review
- Analytical and clinical evaluation of a novel real-time PCR-based detection kit for Mpox virus.Medical microbiology and immunology · 2024Article
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Authors and funding
11 authors at 2 institutions in 1 country.
Funding
Abstract
backgroundIn spring of 2022, an outbreak of monkeypox (mpox) spread worldwide. Here, we describe performance characteristics of monkeypox virus (MPXV)-specific and pan-orthopoxvirus qPCR assays for clinical use.
methodsWe validated probe-based qPCR assays targeting MPXV-specific loci F3L and G2R (genes MPXVgp052/OPG065 and MPXVgp002 and gp190/OPG002, respectively) and a pan-orthopoxvirus assay targeting the E9L locus (MPXVgp057/OPG071). Clinical samples and synthetic controls were extracted using the Roche MP96 or Promega Maxwell 48 instrument. qPCR was performed on the AB7500 thermocycler. Synthetic control DNA and high concentration clinical samples were quantified by droplet PCR. Cross-reactivity was evaluated for camelpox and cowpox genomic DNA, vaccinia culture supernatant, and HSV- and VZV-positive clinical specimens. We also tested the performance of the F3L assay using dry swabs, Aptima vaginal and rectal swabs, nasopharyngeal, rectal, and oral swabs, cerebrospinal fluid, plasma, serum, whole blood, breastmilk, urine, saliva, and semen.
resultsThe MPXV-F3L assay is reproducible at a limit of detection (LoD) of 65.6 copies/mL of viral DNA in viral transport medium/universal transport medium (VTM/UTM), or 3.3 copies/PCR reaction. No cross-reactivity with herpesviruses or other poxviruses was observed. MPXV-F3L detects MPXV DNA in alternative specimen types, with an LoD ranging between 260-1000 copies/mL, or 5.7-10 copies/PCR reaction. In clinical swab VTM specimens, MPXV-F3L and MPXV-G2R assays outperformed OPXV-E9L by an average of 2.4 and 2.8 Cts, respectively. MPXV-G2R outperformed MPXV-F3L by 0.4 Cts, consistent with presence of two copies of G2R present in labile inverted terminal repeats (ITRs) of MPXV genome.
conclusionsMPXV is readily detected by qPCR using three clinically validated assays.
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