ABSTRACT.Amoebic liver abscess (ALA) is a significant public health concern in tropical countries. Traditional diagnostic methods, such as microscopy, have low sensitivity, and nested polymerase chain reaction (nPCR) has lengthy turnaround times. In this study, we aimed to evaluate the effectiveness of the recombinase polymerase amplification (RPA) assay for detecting Entamoeba histolytica in amoebic liver pus samples. The assay was tested on 150 clinical pus samples collected from suspected ALA patients in the emergency department of a tertiary care center located in Chandigarh, northern India. For comparison with RPA, nPCR was also performed on these samples. Of 150 samples, 79 (53%) tested positive for E. histolytica using the RPA assay, and similar (79; 53%) number of samples tested positive with nested PCR. In addition, sequences obtained through Sanger sequencing showed high similarity with other sequences in the National Center for Biotechnology Information database and were submitted to the database. Our findings highlight the potential of RPA as a valuable tool for the accurate diagnosis of ALA.