Comparative Analysis of ELISA, MAT, and RT-PCR for Early Diagnosis of Human Leptospirosis

Abstract

A neglected zoonotic illness that affects people all around the world is human leptospirosis. Contact with soil or water tainted by sick animals' urine is the primary way it spreads. Early identification is challenging since the symptoms frequently mimic those of other feverish diseases including dengue, malaria, or influenza. This study evaluated the effectiveness of the Enzyme-Linked Immunosorbent Assay (ELISA), Microscopic Agglutination Test (MAT), and Real-Time Polymerase Chain Reaction (RT-PCR) in a structured diagnostic process for suspected human leptospirosis. Blood samples were taken under sterile conditions and analyzed for both serological and molecular details. ELISA was used to detect IgM and IgG antibodies. MAT served as the serological standard, while RT-PCR targeted the LipL32 gene for direct detection of the pathogen. Clinical, demographic, and laboratory data were compiled into a standard database. After that, the data undergo preprocessing to ensure consistency. The study assessed diagnostic performance using Receiver Operating Characteristic (ROC) analysis. The Area under Curve (AUC) values were 0.942 for MAT, 0.904 for ELISA, and 0.981 for RT-PCR. Among these, RT-PCR showed the best diagnostic accuracy, especially within the first 7 days of illness, proving its advantage for early detection. These results highlight the supportive role of RT-PCR alongside serological tests, which aids in the quick, sensitive, and reliable diagnosis of leptospirosis.