Kumar D, Chauhan T K S, Agarwal R K, Dhama K, Goswami P P, Mariappan A K, Tiwari A K, Mishra B P
Division of Veterinary Biotechnology, Indian Veterinary Research Institute, Izatnagar, 243122, UP, India.
Division of Livestock Product Technology, Indian Veterinary Research Institute, Izatnagar, 243122, UP, India.
Arch Virol. 2017 Apr;162(4):979-985. doi: 10.1007/s00705-016-3184-1. Epub 2016 Dec 24.
We applied a probe-based real-time loop-mediated isothermal amplification (Cy5-RTqLAMP) technique targeting the avian reovirus (ARV) S3 gene to develop a rapid, sensitive, and specific method for virus detection and quantification. This test specifically detected the presence of ARV, but not other viruses or bacteria present in clinical or artificially spiked samples, including Newcastle disease virus, infectious bursal disease virus, fowl adenovirus, Marek's disease virus, Escherichia coli, and Salmonella spp. This test can detect ARV in less than one hour with an analytical sensitivity of 10 viral gene copies and 1 fg of total cDNA. The Cy5-RTqLAMP does not yield false positive results and is 100 times more sensitive than conventional PCR. This test was shown to be able to detect the presence of ARV in clinical samples. A similar strategy may be used for detection of other important human and animal viral pathogens.
我们应用了一种基于探针的实时环介导等温扩增技术（Cy5-RTqLAMP），该技术靶向禽呼肠孤病毒（ARV）S3基因，以开发一种用于病毒检测和定量的快速、灵敏且特异的方法。该检测方法能特异性检测到ARV的存在，但对临床样本或人工加样样本中存在的其他病毒或细菌无效，这些病毒和细菌包括新城疫病毒、传染性法氏囊病病毒、禽腺病毒、马立克氏病病毒、大肠杆菌和沙门氏菌属。该检测方法可在不到一小时内检测到ARV，分析灵敏度为10个病毒基因拷贝和1 fg的总cDNA。Cy5-RTqLAMP不会产生假阳性结果，且比传统PCR灵敏100倍。该检测方法已被证明能够检测临床样本中ARV的存在。类似的策略可用于检测其他重要的人类和动物病毒病原体。
