High Resolution Melting (HRM) approach-based strategy for simultaneous differentiation of five Babesia species response for human babesiosis

Abstract

Abstract Background Five Babesia species, including Babesia microti, Babesia divergens, Babesia duncani, Babesia motasi, and Babesia crassa-like, are considered to be the major pathogens responsible for human babesiosis in endemic regions. Each of these species possesses a variable degree of virulence towards its hosts. Therefore, the accurate identification of these species is critical for the adoption of appropriate therapies. Methods In this study, we developed a real-time PCR high-resolution melting analysis (qPCR- HRM) approach targeting 18S rRNA for distinguishing the five Babesia spp. based on melting temperature (Tm) and genotype confidence percent (GCP) values. This approach was evaluated using 429 blood samples from patients with a history of tick bites and 200 positive specimens from laboratory infections. Results The results showed that the sensitivity and specificity of the proposed qPCR-HRM based method were 95% and 100%, respectively, and the detection limit weas 1-100 copies of plasmid. The detection performance of the assay was assessed using clinical and laboratory infected specimens. Conclusions The developed detection method provides a useful tool for the epidemiological investigation of human babesiosis and pre-transfusion screening.