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Detection of bovine Babesiosis in Mozambique by a novel seminested hot-start PCR method.

机译:新型半巢式热启动PCR方法检测莫桑比克的牛杆状杆菌病。

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摘要

Babesiosis is a tick borne disease (TBD) caused by parasites of the genus Babesia, with considerable worldwide economic, medical, and veterinary impact. Bovine babesiosis and other TBDs were considered responsible for 50% of the deaths of cattle that occurred in Mozambique in the first year after importation from neighbouring countries.Here, we present the detection of Babesia bigemina and Babesia bovis in cattle from Mozambique using two distinct PCR methods. For this study, blood samples were collected in one farm located near Maputo city. The DNA samples were analyzed using a previously described nested PCR and a novel hot-start PCR method. Primers were selected for the hot-start PCR based on the putative gene of an undescribed aspartic protease named babesipsin, present in both B. bovis and B. bigemina. The combination of hot-start polymerase and long primers (29–31 bp) were in this study determinant for the successful amplification and detection in only one PCR. With a seminested approach the sensitivity was further increased. The babesipsin seminested hot-start PCR was in this study more sensitive than the nested PCR. A total of 117 field samples were tested by seminested hot-start PCR, and 104 were positive for B. bigemina (90%), 97 were positive for B. bovis (82%), 86 were mixed infections (52%) and only 2 were negative for both Babesia species (1.7%). The results confirm that this area of Mozambique is endemic for babesiosis, and that this TBD should be regarded as a threat for imported cattle.
机译:巴贝氏病是由巴贝氏菌属的寄生虫引起的tick传播疾病(TBD),在全球范围内对经济,医学和兽医产生重大影响。从邻国进口后第一年,牛巴贝虫病和其他TBD被认为是造成莫桑比克牛死亡的50%。在这里,我们介绍了使用两种不同的PCR方法在莫桑比克牛中检测到大巴贝斯虫和牛巴贝斯虫方法。在这项研究中,从位于马普托市附近的一个农场采集了血液样本。使用先前描述的巢式PCR和新型热启动PCR方法分析DNA样品。基于未描述的天冬氨酸蛋白酶(称为牛肝素)的推定基因选择引物用于热启动PCR,该基因存在于牛双歧杆菌和双歧杆菌中。在本研究中,热启动聚合酶和长引物(29-31 bp)的组合是仅一次PCR成功扩增和检测的决定因素。使用半嵌套方法,可以进一步提高灵敏度。巴贝西平半巢式热启动PCR在本研究中比巢式PCR更为敏感。通过半巢式热启动PCR测试了总共117个现场样品,其中104个为双歧杆菌阳性(90%),97个为牛双歧杆菌阳性(82%),86个为混合感染(52%),仅两种巴贝斯虫属均为阴性(1.7%)。结果证实,莫桑比克的这一地区是巴贝西斯病的地方病,该TBD应被视为对进口牛的威胁。

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