Analysis of PCR Techniques for Detection of Leptospira: A Narrative Review

Leptospirosis, a widespread zoonotic disease caused by pathogenic Leptospira species, is recognised as posing significant challenges to both public health and veterinary sectors. Traditional diagnostic methods, including serological assays and culture techniques, are often characterised by limited sensitivity, particularly during the early stages of infection. In contrast, enhanced accuracy and rapid detection capabilities have been demonstrated by molecular diagnostics- especially Polymerase Chain Reaction (PCR)-based approaches. In this review, the development and application of various PCR-based methods, including conventional PCR, nested PCR, quantitative real-time PCR (qPCR), digital PCR, and Loop-Mediated Isothermal Amplification (LAMP), have been explored for detecting Leptospira in clinical and environmental samples. These technologies have been shown to not only improve sensitivity and specificity but also facilitate early and reliable diagnosis. A comprehensive literature search was conducted using PubMed, Scopus, and Web of Science, focusing on studies published between 2000 and 2025. The selected literature has highlighted the diagnostic strengths, limitations, and practical considerations associated with each method. Furthermore, the future integration of multiplex PCR assays and portable diagnostic tools is discussed to expand point-of-care testing, particularly in resource-limited settings. Standardisation of protocols and cross-platform validation are emphasised as critical for achieving widespread, effective use. Overall, leptospirosis diagnostics continue to be redefined by PCR-based strategies, supporting improved disease surveillance and control.