Parida M M
Department of Virology, Defence R and D Establishment, Gwalior 474 002, India.
J Biosci. 2008 Nov;33(4):617-28. doi: 10.1007/s12038-008-0079-7.
The development of technologies with rapid and sensitive detection capabilities and increased throughput have become crucial for responding to greater number threats posed by emerging and re-emerging viruses in the recent past. The conventional identification methods require time-consuming culturing, and/or detection of antibodies,which are not very sensitive and specific. The recent advances in molecular biology techniques in the field of genomics and proteomics greatly facilitate the rapid identification with more accuracy. We have developed two real-time assays ie., SYBR green I based real time reverse transcription polymerase chain reaction (RT-PCR) and RT-loop-mediated isothermal amplification (LAMP) assay for rapid detection as well as typing of some of the emerging viruses of biomedical importance viz. dengue, Japanese encephalitis, chikungunya, west Nile, severe acute respiratory syndrome virus (SARS) etc. Both these techniques are capable of detection and differentiation as well as quantifying viral load with higher sensitivity, rapidity, specificity. One of the most important advantages of LAMP is its field applicability, without requirement of any sophisticated equipments. Both these assays have been extensively evaluated and validated with clinical samples of recent epidemics from different parts of India. The establishment of these real time molecular assays will certainly facilitate the rapid detection of viruses with high degree of precision and accuracy in future.
近年来,开发具有快速灵敏检测能力和更高通量的技术对于应对新出现和再次出现的病毒带来的更多威胁至关重要。传统的鉴定方法需要耗时的培养和/或抗体检测,这些方法不太灵敏且特异性不强。基因组学和蛋白质组学领域分子生物学技术的最新进展极大地促进了更准确的快速鉴定。我们开发了两种实时检测方法,即基于SYBR Green I的实时逆转录聚合酶链反应(RT-PCR)和RT环介导等温扩增(LAMP)检测方法,用于快速检测以及对一些具有生物医学重要性的新出现病毒进行分型,即登革热、日本脑炎、基孔肯雅热、西尼罗河病毒、严重急性呼吸综合征病毒(SARS)等。这两种技术都能够进行检测、区分以及以更高的灵敏度、速度和特异性对病毒载量进行定量。LAMP最重要的优点之一是其现场适用性,无需任何复杂设备。这两种检测方法都已通过来自印度不同地区近期疫情的临床样本进行了广泛评估和验证。这些实时分子检测方法的建立肯定会在未来促进对病毒的高精度和准确快速检测。