Detection of Babesia gibsoni in dogs by combining recombinase polymerase amplification (RPA) with lateral flow (LF) dipstick

• Published in: Parasitology research (1987) Vol. 117; no. 12; pp. 3945 - 3951
• Main Authors: Cui, Jie, Zhao, Yangnan, Sun, Yali, Yu, Long, Liu, Qin, Zhan, Xueyan, Li, Muxiao, He, Lan, Zhao, Junlong
• Format: Journal Article
• Language: English
• Published: Berlin/Heidelberg Springer Berlin Heidelberg 01.12.2018; Springer Nature B.V
• Subjects: Animals; Babesia - genetics; Babesia - isolation & purification; Babesia gibsoni; Babesiosis; Babesiosis - diagnosis; Babesiosis - parasitology; Biomedical and Life Sciences; Biomedicine; China; Cytochrome-c oxidase; Dog Diseases - diagnosis; Dog Diseases - parasitology; Dogs; Immunology; Medical Microbiology; Microbiology; Nucleic Acid Amplification Techniques - methods; Original Paper; Parasites; Polymerase chain reaction; Polymerase Chain Reaction - methods; Protozoa; Recombinase; Recombinases - genetics; RNA, Ribosomal, 18S - genetics; rRNA 18S; Sensitivity and Specificity; China; Babesiosis; Lateral flow; Diagnosis; Recombinase polymerase amplification; Babesia gibsoni
• DOI: 10.1007/s00436-018-6104-3
• ISSN: 0932-0113

Babesia gibsoni is a protozoan parasite responsible for the majority of reported cases of canine babesiosis in China. Currently, microscopic examination of the Giemsa-stained thin blood smears is the main diagnosis method in clinic. Here, we report the recombinase polymerase amplification-lateral flow (LF-RPA) dipstick detection method for targeting B. gibsoni cytochrome c oxidase subunit I ( cox I) gene. The reaction takes only 20–30 min under isothermal temperatures between 30 and 45 °C. Specificity was evaluated using DNA from related apicomplexan parasites and their host, while the sensitivity was calculated based on the DNA from the experimental B. gibsoni- infected dogs. Results indicated that the LF-RPA method is 20 times more sensitive than the conventional PCR based on 18S rRNA and has no cross reaction with any other test DNAs. The applicability of the LF-RPA method was further evaluated using 15 samples collected from clinic. Thirteen of the 15 samples (86.67%) were detected as positive by LF-RPA, while 10 of them (66.67%) were found positive by conventional PCR. Overall, the novel LF-RPA assay is effective for the detection of B. gobsini and has considerable advantages over the conventional PCR in sensitivity, specificity, simplicity in operation, less time consumption, and visual detection. The LF-RPA method may facilitate the surveillance and early detection of B. gibsoni infection in dogs.