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Detection and quantitation of Anaplasma marginale in carrier cattle by using a nucleic acid probe.

机译:使用核酸探针检测和定量测定牛携带的无性无形体。

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摘要

Cattle which have recovered from acute infection with Anaplasma marginale, a rickettsial hemoparasite of cattle, frequently remain persistently infected with a low-level parasitemia and serve as reservoirs for disease transmission. To fully understand the role of these carriers in disease prevalence and transmission, it is essential that low levels of parasitemia can be accurately detected and quantitated. We have developed a nucleic acid probe, derived from a portion of a gene encoding a 105,000-molecular-weight surface protein, that can detect A. marginale-infected erythrocytes. The probe is specific for A. marginale and can detect 0.01 ng of genomic DNA and 500 to 1,000 infected erythrocytes in 0.5 ml of blood, which is equivalent to a parasitemia of 0.000025%. This makes the probe at least 4,000 times more sensitive than light microscopy. Hybridization of the probe with treated blood from animals proven to be carriers of anaplasmosis showed that parasitemia levels were highly variable among carriers, ranging from greater than 0.0025 to less than 0.000025%. Parasitemia levels of individual animals on different dates were also variable. These results imply that, at any given time, individuals within a group of cattle may differ significantly in their abilities to transmit disease.
机译:从牛无性无浆膜(一种牛的rick病性血液寄生虫)的急性感染中恢复的牛,经常经常持续感染低水平的寄生虫病,并作为疾病传播的库。为了充分了解这些携带者在疾病流行和传播中的作用,至关重要的是,必须准确检测和定量低水平的寄生虫病。我们已经开发了一种核酸探针,该探针源自编码105,000分子量表面蛋白的基因的一部分,可以检测出被A.marginale感染的红细胞。该探针对margin曲霉具有特异性,可以检测0.5毫升血液中的0.01 ng基因组DNA和500到1,000个感染的红细胞,这相当于0.000025%的寄生虫病。这使探头的灵敏度至少比光学显微镜高4000倍。探针与被证明是厌氧菌携带者的动物的经处理的血液的杂交表明,携带者中的寄生虫血症水平变化很大,范围从大于0.0025到小于0.000025%。个别动物在不同日期的寄生虫血症水平也不同。这些结果表明,在任何给定时间,一组牛群中的个体传播疾病的能力可能存在显着差异。

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