Surra, caused by Trypanosoma (T.) evansi, causes significant economic losses in many parts of the world including Indonesia. Unfortunately, T. evansi is very difficult to distinguish morphologically from other members of the Trypanozoon subgenus (T. equiperdum and T. brucei sensu lato). Molecular identification using polymerase chain reaction (PCR) with minicircle primers followed by maxicircle primers is a possible approach to be applied in Trypanozoon endemic area such as Asia and Africa. However, PCR is not suitable for application in laboratories with limited resources. The recombinase polymerase amplification (RPA) technique is a promising alternative to PCR for resource-constrained laboratories or point-of-care (POC) settings. In this study, we compared the diagnostic capabilities of RPA and PCR. To achieve that, a total of 39 isolates, comprising 12 isolates of T. evansi and 27 isolates of T. equiperdum, were tested using PCR and RPA. RPA and PCR had an agreement coefficient of >0.9, categorized as a very good agreement according to Altman’s criteria. The comparison between RPA and PCR showed an agreement of 97.4 and 98.7% for identification and detection, respectively, while the level of detection of RPA and PCR was 10¹ trypanosomes/mL. These results indicate that RPA is fundamentally promising technique for detection of Trypanozoon members in laboratories with limited facilities as an alternative to PCR.

To Cite This Article: Subekti DT, Santoso ZA, Suwanti LT, Sunarno S, Mufasirin M and Kurniawati DA, 2025. Comparison of Recombinase Polymerase Amplification with Polymerase Chain Reaction for Species Identification of Trypanozoon Members Based on Minicircle and Maxicircle Genes. Pak Vet J. http://dx.doi.org/10.29261/pakvetj/2025.006