Improved Molecular Tools for Detection of Babesia bigemina

ROMINA PETRIGH; PAULA RUYBAL; CAROLINA THOMPSON; ROBERTO NEUMANN; ROSALIA MORETTA; SILVINA WILKOWSKY; GRACIELA DRAGHI; IGNACIO ECHAIDE; SUSANA TORIONI DE ECHAIDE; MARISA FARBER

ANNALS OF THE NEW YORK ACADEMY OF SCIENCES.

Wiley Blackwell

Año: 2008 vol. 1149 p. 155 - 157

0077-8923

Molecular detection of Babesia bigemina involves a nested PCR protocol and reverseline blot hybridization (RLBH) assay based on the 18S gene. In this study, we report thedevelopment ofmolecular tools for improving B. bigemina detection in bovine blood—aone-step PCR assay based on the amplification of rap-1a paralogous and a new RLBHBabesia spp. 18S probe. The one-step PCR assay is highly specific, with an estimatedanalytical sensitivity corresponding to 0.00002% parasitemia. The RLBH assay, with anewB. bigemina probe, allows the detection of all tested B. bigemina isolates showing nocross-hybridization with B. bovis 18S gene. By developing this highly specific and sensitiveone-step PCR and upgrading the RLBH assay for B. bigemina, we have improvedmolecular assays which, together with serologic methods, provide valuable tools forepidemiologic studies of bovine babesiosis.