Petrigh Romina, Ruybal Paula, Thompson Carolina, Neumann Roberto, Moretta Rosalia, Wilkowsky Silvina, Draghi Graciela, Echaide Ignacio, de Echaide Susana Torioni, Farber Marisa
INTA-Castelar, CC25, CP 1712 Castelar, Buenos Aires, Argentina.
Ann N Y Acad Sci. 2008 Dec;1149:155-7. doi: 10.1196/annals.1428.038.
Molecular detection of Babesia bigemina involves a nested PCR protocol and reverse line blot hybridization (RLBH) assay based on the 18S gene. In this study, we report the development of molecular tools for improving B. bigemina detection in bovine blood-a one-step PCR assay based on the amplification of rap-1a paralogous and a new RLBH Babesia spp. 18S probe. The one-step PCR assay is highly specific, with an estimated analytical sensitivity corresponding to 0.00002% parasitemia. The RLBH assay, with a new B. bigemina probe, allows the detection of all tested B. bigemina isolates showing no cross-hybridization with B. bovis 18S gene. By developing this highly specific and sensitive one-step PCR and upgrading the RLBH assay for B. bigemina, we have improved molecular assays which, together with serologic methods, provide valuable tools for epidemiologic studies of bovine babesiosis.
双芽巴贝斯虫的分子检测涉及一种基于18S基因的巢式PCR方案和反向线印迹杂交（RLBH）分析。在本研究中，我们报告了用于改进牛血液中双芽巴贝斯虫检测的分子工具的开发——一种基于rap - 1a旁系同源物扩增的一步PCR分析以及一种新的巴贝斯虫属18S探针的RLBH分析。一步PCR分析具有高度特异性，估计分析灵敏度对应于0.00002%的虫血症。使用新的双芽巴贝斯虫探针的RLBH分析能够检测所有测试的双芽巴贝斯虫分离株，且与牛巴贝斯虫18S基因无交叉杂交。通过开发这种高度特异性和灵敏的一步PCR并升级双芽巴贝斯虫的RLBH分析，我们改进了分子检测方法，这些方法与血清学方法一起，为牛巴贝斯虫病的流行病学研究提供了有价值的工具。
