Development and evaluation of two nested PCR assays for the detection of Babesia bovis from cattle blood
Development and evaluation of two nested PCR assays for the detection of Babesia bovis from cattle blood
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DOI：
10.1016/j.vetpar.2010.04.011
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发表时间：
2010-08-27
影响因子：
2.6
通讯作者：
Igarashi, Ikuo
中科院分区：
文献类型：
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作者：
AbouLaila, Mahmoud;Yokoyama, Naoaki;Igarashi, Ikuo
本研究利用B巴氏杆菌膜蛋白基因BBOV IV 005650（BV 5650）和BBO_IV 008970（BV 8970），建立了两种牛巴氏杆菌感染的巢式nPCR诊断方法，并与B巴氏杆菌Rhoptery相关蛋白1基因（RAP-1）的nPCR方法进行了比较。nPCR对体外培养的牛B弓形虫的敏感性分别为10（-8）%、10（-6）%和10（-7）%，其中BV 5650、BV 8970和RAP-1的敏感性分别为10（-8）%、10（-6）%和10（-7）%。对于BV 5650和BV 8970和RAP，nPCR分别检测到每次测试低至1fg的基因组DNA和100 fg的基因组DNA。BV 5650 nPCR检测牛B的灵敏度最高，BV 5650 nPCR检测牛B的灵敏度最高，BV 5650 nPCR检测牛B的灵敏度最高，BV 5650 nPCR检测牛B的灵敏度最高。用于全球牛B bovis感染实验室诊断评估的诊断工具（C）2010 Elsevier B V保留所有权利
We developed and evaluated two nested polymerase chain reaction (nPCR) assays for the diagnosis of Babesia bovis infection in cattle based on two membrane protein genes from B bows BBOV IV005650 (BV5650) and BBO_IV008970 (BV8970) The specificities and sensitivities of the tests were compared with B bovis Rhoptery associated protein 1 gene (RAP-1) nPCR. The specificity of the tests was 100% for B bovis DNA The sensitivities of nPCR to B bows from the in vitro cultured parasites were as low as 10(-8)% 10(-6)% and 10(-7)% parasitemia for BV5650 BV8970 and RAP-1 nPCR, respectively The nPCR detected as little as 1 fg genomic DNA per test for BV5650 and 100 fg per test for both BV8970 and RAP-1 genes For field applications the sensitivity was evaluated to a total of 165 field samples from Ghana Mongolia Brazil and Japan The nPCR assay of BV5650 was the most sensitive for the detection of B bovis from the field samples The BV5650 nPCR assay provides a good diagnostic tool for laboratory diagnostic assessment of B bovis infection in cattle worldwide (C) 2010 Elsevier B V All rights reserved