INTRODUCTION: Ensuring accuracy in the diagnosis of leishmaniasis is crucial due to the myriad of potential differential diagnoses. Given the inherent limitations of serological techniques, real-time polymerase chain reaction (qPCR) emerges as a superior alternative. Furthermore, parasitological methods, conventionally regarded as the gold standard owing to their high specificity, encounter challenges concerning sensitivity and invasiveness for patients. In this context, the present study aims to assess, via meta-analysis, the performance of qPCR in diagnosing visceral and cutaneous leishmaniasis. METHOD: This meta-analysis encompassed studies published between January 2011 and December 2022, sourced from six databases (PubMed, LILACS, Scopus, Scielo, EMBASE, and Web of Science), utilizing the keywords "qPCR," "molecular diagnosis," and "leishmaniasis." Epidemiological studies focusing on the efficacy of qPCR for leishmaniasis diagnosis were included. Data such as study demographics, geographic locations, sampling techniques, and the number of positive qPCR results were aggregated and analyzed to derive overall positivity rates, sensitivity, and specificity values associated with qPCR. Heterogeneity analysis was conducted on the data to select appropriate models, and the collective efficacy data of qPCR were illustrated in forest plots. RESULTS: Fifty-four studies met all inclusion criteria. The positivity rates for human visceral and cutaneous leishmaniasis were 27.07% (95% CI: 17.81-36.33%) and 60.40% (95% CI: 30.23-90.57%), respectively. In cases of visceral leishmaniasis in dogs, cats, and wild animals, the positivity rates were 26.55% (95% CI: 21.40-31.70%), 0.92% (95% CI: 0.09-1.75%), and 28.98% (95% CI: 21.86-35.10%), respectively. Analysis of the selected studies revealed high overall sensitivity and specificity values achieved with qPCR, at 91.08% (95% CI: 81.77-100.39%) and 98.08% (95% CI: 97.13-99.03%), respectively. CONCLUSION: This study indicates that qPCR is a highly sensitive and specific tool, adequately suitable for the diagnosis of human visceral and cutaneous leishmaniasis, as well as visceral leishmaniasis in animals.