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用于诊断锥蝽(半翅目:猎蝽科)物种中锥蝽病毒(TrV)的AC-ELISA和RT-PCR检测方法。

AC-ELISA and RT-PCR assays for the diagnosis of triatoma virus (TrV) in triatomines (Hemiptera: Reduviidae) species.

作者信息

Marti Gerardo A, González Ester T, García Juan J, Viguera Ana R, Guérin Diego M A, Echeverría María G

机构信息

Centro Regional de Investigaciones Científicas y Transferencias Tecnológicas La Rioja (CRILAR, CONICET) Entre Ríos and Mendoza, 5310, Anillaco, La Rioja, Argentina.

出版信息

Arch Virol. 2008;153(8):1427-32. doi: 10.1007/s00705-008-0130-x. Epub 2008 Jun 19.

Abstract

Triatoma virus (TrV) is the only entomopathogenic virus found in triatomines. TrV replicates in cells of the midgut epithelium of triatomines, causing a high mortality rate and delayed development of the infected insect. In this work, we report an antigen-capture enzyme-linked immunosorbent assay (AC-ELISA) and a reverse transcription-polymerase chain reaction (RT-PCR) assay for detection of TrV infection. For antiserum production, rabbits and hens where inoculated with purified TrV. Antiserum reactivity was checked by immunodiffusion, and its specificity was confirmed by western blot and AC-ELISA. Totally 90 fecal samples from T. infestans were analysed. AC-ELISA and RT-PCR results correlated well with transmission electron microscopy (EM) observations, which are considered the gold standard, with Kappa values of 0.73 for AC-ELISA and 0.93 for RT-PCR when compared with EM. Applications and complementary uses of the two techniques reported in this work are discussed.

摘要

锥蝽病毒(TrV)是在锥蝽中发现的唯一一种昆虫致病病毒。TrV在锥蝽中肠上皮细胞中复制,导致被感染昆虫的死亡率很高且发育延迟。在这项研究中,我们报告了一种用于检测TrV感染的抗原捕获酶联免疫吸附测定(AC-ELISA)和逆转录聚合酶链反应(RT-PCR)测定。为了制备抗血清,用纯化的TrV对兔子和母鸡进行接种。通过免疫扩散检查抗血清反应性,并通过蛋白质印迹和AC-ELISA确认其特异性。总共分析了90份来自骚扰锥蝽的粪便样本。AC-ELISA和RT-PCR结果与被视为金标准的透射电子显微镜(EM)观察结果相关性良好,与EM相比,AC-ELISA的Kappa值为0.73,RT-PCR的Kappa值为0.93。本文还讨论了所报道的这两种技术的应用及互补用途。

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