Table 2.

Overview of diagnostic methods for the detection of monkeypox virus

Method	Description	Strengths	Limitations	References
Polymerase chain reaction (PCR)	Molecular detection of MPXV DNA, targeting specific genes	High sensitivity and specificity; gold standard	Requires advanced labs and trained personnel	[76]
Next-generation sequencing (NGS)	Comprehensive genomic sequencing of MPXV for clade identification	Detects mutations; tracks viral evolution	High cost; complex analysis; time-intensive	[77, 78]
serology (ELISA)	Detects antibodies against MPXV in patient samples	Useful for epidemiological studies	Cross-reactivity with other Orthopoxviruses; limited for acute diagnosis	[79]
Rapid diagnostic tests (RDTs)	Lateral flow assays for antigen or antibody detection	Portable; quick results	Lower sensitivity compared to PCR	[80, 81]
Loop-Mediated Isothermal Amplification (LAMP)	Isothermal amplification of MPXV DNA	Portable; fast; field deployable	Lower sensitivity and specificity than PCR	[80, 81]
Clinical diagnosis	Based on characteristic symptoms, such as fever, rash, and lymphadenopathy	Useful in resource-limited settings	Overlaps with other illnesses; relies on clinician expertise	[82]
Viral culture	Isolation of live virus in cell culture	Confirms live virus presence	Requires BSL-3 labs; time-consuming	[82]
CRISPR-based assays	Uses CRISPR-Cas systems for precise nucleic acid detection	Highly specific, portable	Emerging technology; limited availability	[83, 84]
Recombinase polymerase amplification (RPA)	Isothermal DNA amplification at ~ 37–42 °C; results in ~ 15–20 min	Ultra-rapid; portable; suitable for low-resource settings	Still under optimization; risk of cross-contamination	[85]