Shete Anita M, Yadav Pragya, Kumar Vimal, Nikam Tushar, Mehershahi Kurosh, Kokate Prasad, Patil Deepak, Mourya Devendra T
Maximum Containment Laboratory, ICMR-National Institute of Virology, Pune, India.
Indian J Med Res. 2017 Jan;145(1):90-96. doi: 10.4103/ijmr.IJMR_1447_15.
BACKGROUND & OBJECTIVES: Bats are recognized as important reservoirs for emerging infectious disease and some unknown viral diseases. Two novel viruses, Malsoor virus (family Bunyaviridae, genus, Phlebovirus) and a novel adenovirus (AdV) (family, Adenoviridae genus, Mastadenovirus), were identified from Rousettus bats in the Maharashtra State of India. This study was done to develop and optimize real time reverse transcription - polymerase chain reaction (RT-PCR) assays for Malsoor virus and real time and nested PCR for adenovirus from Rousettus bats.
For rapid and accurate screening of Malsoor virus and adenovirus a nested polymerase chain reaction and TaqMan-based real-time PCR were developed. Highly conserved region of nucleoprotein gene of phleboviruses and polymerase gene sequence from the Indian bat AdV isolate polyprotein gene were selected respectively for diagnostic assay development of Malsoor virus and AdV. Sensitivity and specificity of assays were calculated and optimized assays were used to screen bat samples.
Molecular diagnostic assays were developed for screening of Malsoor virus and AdV and those were found to be specific. Based on the experiments performed with different parameters, nested PCR was found to be more sensitive than real-time PCR; however, for rapid screening, real-time PCR can be used and further nested PCR can be used for final confirmation or in those laboratories where real-time facility/expertise is not existing.
INTERPRETATION & CONCLUSIONS: This study reports the development and optimization of nested RT-PCR and a TaqMan-based real-time PCR for Malsoor virus and AdV. The diagnostic assays can be used for rapid detection of these novel viruses to understand their prevalence among bat population.
蝙蝠被认为是新发传染病及一些未知病毒性疾病的重要宿主。在印度马哈拉施特拉邦的果蝠中鉴定出两种新型病毒，马尔苏尔病毒（布尼亚病毒科白蛉病毒属）和一种新型腺病毒（腺病毒科 Mastadenovirus 属）。本研究旨在开发并优化用于检测果蝠中马尔苏尔病毒的实时逆转录 - 聚合酶链反应（RT-PCR）检测方法以及用于腺病毒的实时和巢式PCR检测方法。
为快速准确地筛查马尔苏尔病毒和腺病毒，开发了巢式聚合酶链反应和基于TaqMan的实时PCR。分别选择白蛉病毒核蛋白基因的高度保守区域以及印度蝙蝠腺病毒分离株多聚蛋白基因的聚合酶基因序列，用于开发马尔苏尔病毒和腺病毒的诊断检测方法。计算检测方法的敏感性和特异性，并使用优化后的检测方法筛查蝙蝠样本。
开发了用于筛查马尔苏尔病毒和腺病毒的分子诊断检测方法，且这些方法具有特异性。基于不同参数进行的实验，发现巢式PCR比实时PCR更敏感；然而，为了快速筛查，可以使用实时PCR，进一步的巢式PCR可用于最终确认，或者在没有实时检测设备/专业技术的实验室中使用。
本研究报告了用于马尔苏尔病毒和腺病毒的巢式RT-PCR及基于TaqMan的实时PCR的开发与优化。这些诊断检测方法可用于快速检测这些新型病毒，以了解它们在蝙蝠种群中的流行情况。
