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用于检测狂犬病病毒的逆转录聚合酶链反应-酶联免疫吸附测定法的诊断评估

Diagnostic evaluation of RT-PCR-ELISA for the detection of rabies virus.

作者信息

Aravindhbabu R P, Manoharan S, Ramadass P

机构信息

Rabies Laboratory, Department of Animal Biotechnology, Madras Veterinary College, Chennai, 600007 India ; National Institute for Animal Biotechnology, 1-121/1, 4th Floor, Axis Clinicals Building, Miyapur, Hyderabad, 500049 India.

Rabies Laboratory, Department of Animal Biotechnology, Madras Veterinary College, Chennai, 600007 India.

出版信息

Virusdisease. 2014 Jan;25(1):120-4. doi: 10.1007/s13337-013-0184-6. Epub 2013 Nov 30.

Abstract

Rabies is primarily a disease of terrestrial and airborne mammals. In most cases, rabies is diagnosed primarily on the basis of clinical symptoms and signs, and a corroborative history of or evidence of an animal bite, death of an animal and incomplete or no vaccination following exposure. The facility for laboratory diagnosis and confirmation of rabies is available in only a few institutions in India. Diagnostic tests using conventional assays like fluorescent antibody test (FAT) are unreliable at times, despite the clinical diagnosis. Currently, there are a number of molecular tests that can be used to complement conventional tests in rabies diagnosis. We have developed and evaluated an RT-PCR-ELISA using a panel of brain tissue samples from rabies suspected animals of various species. This assay was able to detect rabies virus genome in all the 43 samples that were previously tested positive for rabies. Moreover this assay was shown to be 100 % sensitive and specific in detecting the rabies virus genome in post-mortem brain tissue samples from different species of animals. Our pilot study shows the potential of this assay as an alternative diagnostic test when the samples are unsuitable for use in FAT and also a supplementary test to FAT. In addition, the region of nucleoprotein gene amplified using this assay can be used for the molecular investigation of geographical origin of the field strains.

摘要

狂犬病主要是陆生和空中哺乳动物的疾病。在大多数情况下,狂犬病主要根据临床症状和体征以及动物咬伤史、动物死亡情况和暴露后未完全接种或未接种疫苗的佐证来诊断。印度只有少数机构具备狂犬病实验室诊断和确认的设施。尽管有临床诊断,但使用荧光抗体试验(FAT)等传统检测方法进行的诊断测试有时并不可靠。目前,有许多分子检测方法可用于辅助狂犬病的传统诊断测试。我们使用来自各种疑似狂犬病动物的脑组织样本开发并评估了一种逆转录聚合酶链反应 - 酶联免疫吸附测定(RT-PCR-ELISA)。该检测方法能够在之前43个狂犬病检测呈阳性的样本中全部检测到狂犬病病毒基因组。此外,该检测方法在检测不同物种动物死后脑组织样本中的狂犬病病毒基因组时,显示出100%的敏感性和特异性。我们的初步研究表明,当样本不适用于FAT时,该检测方法作为替代诊断测试具有潜力,并且是FAT的补充测试。此外,使用该检测方法扩增的核蛋白基因区域可用于对野外毒株地理来源的分子研究。

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