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抗原捕获酶免疫测定法:与其他检测蜱中斑点热群立克次体方法的比较

Antigen-capture enzyme immunoassay: a comparison with other methods for the detection of spotted fever group rickettsiae in ticks.

作者信息

Radulovic S, Feng H M, Crocquet-Valdes P, Morovic M, Dzelalija B, Walker D H

机构信息

Department of Pathology, University of Texas Medical Branch, Galveston.

出版信息

Am J Trop Med Hyg. 1994 Mar;50(3):359-64. doi: 10.4269/ajtmh.1994.50.359.

Abstract

To evaluate the prevalence of spotted fever group rickettsiae along the Adriatic Coast of Croatia, 832 ticks were examined by hemolymph test, direct immunofluorescence, antigen-capture enzyme immunoassay, and polymerase chain reaction. Very good agreement was observed among direct immunofluorescence, polymerase chain reaction, and antigen-capture enzyme immunoassay. Twelve ticks that were positive by hemolymph test and negative by both direct immunofluorescence and polymerase chain reaction presumably do not represent spotted fever group rickettsiae. By direct immunofluorescence, spotted fever group rickettsiae were present in 12% of Rhipicephalus bursa, 10.6% of Rh. sanguineus, and 7.8% of Dermacentor marginatus. From the 98 ticks containing rickettsia-like organisms by hemolymph test, seven spotted fever group rickettsial isolates were established in cell culture. Four isolates were identified as Rickettsia conorii. The antigen-capture enzyme immunoassay, which utilizes a monoclonal antibody to antigens of the 135-kD surface protein shared among many members of the spotted fever group, is recommended for primary screening of tick samples because it is reliable and yet less labor-intensive than the hemolymph and direct immunofluorescence tests. Although the polymerase chain reaction is too expensive for use as a screening method, it is recommended for confirmation of positive screening results. In addition to the technologic advance of the antigen-capture enzyme immunoassay, this study documented by contemporary methods that R. conorii is present along the eastern Adriatic Coast not only in the classic vector, Rh. sanguineus, but also in Rh. bursa and D. marginatus.

摘要

为评估克罗地亚亚得里亚海沿岸斑点热群立克次体的流行情况,采用血淋巴试验、直接免疫荧光法、抗原捕获酶免疫测定法和聚合酶链反应对832只蜱进行了检测。直接免疫荧光法、聚合酶链反应和抗原捕获酶免疫测定法之间观察到非常好的一致性。12只蜱血淋巴试验呈阳性,但直接免疫荧光法和聚合酶链反应均为阴性,推测这些蜱不携带斑点热群立克次体。通过直接免疫荧光法检测发现,12%的柏氏扇头蜱、10.6%的血红扇头蜱和7.8%的边缘革蜱携带斑点热群立克次体。在98只血淋巴试验检测出含有立克次体样生物体的蜱中,有7株斑点热群立克次体分离株在细胞培养中得以确立。4株分离株被鉴定为康氏立克次体。抗原捕获酶免疫测定法利用针对斑点热群许多成员共有的135-kD表面蛋白抗原的单克隆抗体,推荐用于蜱样本的初步筛查,因为它可靠且比血淋巴试验和直接免疫荧光法所需的劳动力少。虽然聚合酶链反应作为一种筛查方法成本过高,但推荐用于确认筛查阳性结果。除了抗原捕获酶免疫测定法的技术进步外,本研究还通过现代方法证明,康氏立克次体不仅存在于亚得里亚海东部沿岸的经典传播媒介血红扇头蜱中,也存在于柏氏扇头蜱和边缘革蜱中。

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