Abstract
Background: Recently, monkeypox virus (Mpox) infections have been reported from many parts of the world. Mpox can be transmitted through, contact with patients, fomites, sexual contact, infected animals and during pregnancy. Complications of Mpox include encephalitis, pneumonia and ocular infections. Rapid diagnosis is very important for identification of the virus and treatment. Aim: This article is aimed to bring the new knowledge on the laboratory diagnosis of monkeypox. This report deals with sample collection, sample transport and different laboratory techniques available for the diagnosis. Methods: Thorough review of literature was performed with databases Google scholar, EMBASE, PMC, PubMed/Medline, and Cochrane Library, for the period between 1990 to February 2024. Results: Monkeypox (Mpox) is caused by a DNA virus called monkeypox virus (MPXV). Sample collection and transport should be carried out with all biosafety precautions. The laboratory requirements for handling the suspected cases of monkeypox should be conducted in biosafety level-2 labs only. A variety of clinical specimens can be used for the diagnosis of monkeypox. Skin lesions, throat/nasopharyngeal swab, urine, semen, rectal/anal swab, blood and plasma/serum can be collected for the diagnosis. The detection methods are electron microscopy, virus culture using cell lines/embryonated eggs, PCR, RT-PCR and serology. The commonly used techniques are PCR and RT-PCR. Virus culture on monkeypox samples is carried out in CDC laboratory only. Electron microscopy followed by negative staining is performed in reference laboratories only. Conclusions: The detection of monkeypox by molecular methods such as nucleic acid amplification testing is done by conventional PCR, RT- PCR or sequencing. This is the most accurate method to detect monkeypox. The first step is the PCR to detect orthopox virus and the second test is to conduct PCR / sequencing to confirm the monkeypox virus. Very recently, commercial RT-PCR kit to detect monkeypox virus with primers and probes designed to target the F3L gene of monkeypox virus is available.

Key words: monkeypox; Mpox; electron microscopy; RT-PCR; PCR; lab diagnosis.