AbstractBackgroundImproved diagnostic capabilities and accessibility are essential for controlling the outbreak of Human Monkeypox.MethodsWe describe a saliva-based polymerase chain reaction (PCR) assay for Human Monkeypox, in vitro test performance, and clinical implementation of that assay at three testing sites in Los Angeles. Finally, using pre-specified search terms, we conducted a systematic rapid review of PubMed and Web of Science online databases of studies reporting the performance of oral pharyngeal or saliva-based tests for Human Monkeypox.ResultsLaboratory evaluation of the assay showed in silico inclusivity of 100% for 97 strains of Human Monkeypox, with an analytic sensitivity of 250 copies/mL, and 100% agreement compared to known positive and negative specimens. Clinical testing identified 22 cases of Human Monkeypox among 132 individuals (16.7%). Of those 22 cases, 16 (72.7%) reported symptoms, 4 (18.2%) without a rash at the time of testing. Our systematic rapid review identified 6 studies; 23 patients had tests performed on oropharyngeal specimens 100% agreed with the PCR test result of a lesion swab.ConclusionSaliva-based PCR tests are potential tools for outbreak control, and further evaluation of the performance of such tests is warranted.