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Detection of multiple strains of rabies virus RNA using primers designed to target Mexican vampire bat variants.

机译:使用针对墨西哥吸血蝙蝠变异体设计的引物检测狂犬病毒RNA的多种病毒株。

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摘要

A reverse transcription-polymerase chain reaction (RT-PCR), that uses primers specifically designed to amplify a portion of the N gene of vampire bat strains of rabies that circulate in Mexico, but also recognizing most of the rabies variants circulating in endemic areas, was established. This standardized PCR assay was able to detect viral RNA in tenfold serial dilutions up to a 10(7) dilution using stock virus at an original titre of 10(7.5) LD50. The assay was highly specific for rabies virus. Forty different rabies isolates recovered from different species and geographical regions in the country were diagnosed as positive and negative by the fluorescent antibody test (FAT). These same samples were re-examined by both PCR and the mouse inoculation test (MIT). Compared with MIT the PCR exhibited an epidemiological sensitivity of 86% and a specificity of 91% while its positive predictive value was 96%.
机译:逆转录聚合酶链反应(RT-PCR),使用专门设计的引物来扩增在墨西哥传播的狂犬病吸血蝙蝠菌株N基因的一部分,但也可以识别在流行地区传播的大多数狂犬病变体,建立了。使用原始病毒,原始滴度为10(7.5)LD50,这种标准化的PCR检测能够检测到10倍稀释至10(7)稀释度的病毒RNA。该测定对狂犬病毒具有高度特异性。通过荧光抗体试验(FAT),从该国不同物种和地理区域回收的40种不同的狂犬病分离株被诊断为阳性和阴性。通过PCR和小鼠接种试验(MIT)重新检查这些相同的样品。与MIT相比,PCR的流行病学敏感性为86%,特异性为91%,而阳性预测值为96%。

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