COMPARISON OF ANALYTICAL SENSITIVITY AND SPECIFICITY OF REVERSE TRANSCRIPTION LOOP-MEDIATED ISOTHERMAL AMPLIFICATION (RT-LAMP) AND REVERSE TRANSCRIPTION POLYMERASE CHAIN REACTION (RT-PCR) FOR DETECTION OF LOCAL PESTIVIRUS A

Pub Date : 2020-01-01 DOI:10.36899/japs.2020.1.0006
Hammad-ur-Rehman, A. Ahmad, M. Rabbani, M. Tipu
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Abstract

The main aim of this study was to compare reverse transcription loop-mediated isothermal amplification (RT-LAMP) and reverse transcription polymerase chain reaction (RT-PCR) in order to determine their analytical sensitivity and specificity using local characterized Pestivirus A. In order to compare the sensitivity of RT-LAMP and RT-PCR, serial 10 fold dilutions containing 9.91 x 10 to 9.91 x 10 copies of cDNA of Pestivirus A, were prepared and tested. RTLAMP proved more sensitive and was able to detect 9.91x10 copies of cDNA compared to RT-PCR (9.91x10). Both RT-LAMP and RT-PCR were found equally specific as no cross reaction with bovine herpes virus was observed. Present study showed that RT-LAMP assay is highly sensitive, specific, rapid and can be used as an alternative to conventional RT-PCR, for the detection of Pestivirus A.
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逆转录环介导等温扩增(rt-lamp)与逆转录聚合酶链反应(rt-pcr)检测本地鼠疫病毒a型的灵敏度和特异性比较
本研究的主要目的是比较逆转录环介导的等温扩增(RT-LAMP)和逆转录聚合酶链反应(RT-PCR),以确定它们对当地特有的鼠疫病毒A的分析敏感性和特异性。为了比较RT-LAMP和RT-PCR的敏感性,制备了含有9.91 × 10拷贝到9.91 × 10拷贝的鼠疫病毒A cDNA的10倍稀释液并进行了测试。与RT-PCR (9.91x10)相比,RTLAMP更敏感,能够检测到9.91x10个cDNA拷贝。RT-LAMP和RT-PCR均具有相同的特异性,未观察到与牛疱疹病毒的交叉反应。本研究表明,RT-LAMP检测方法灵敏度高、特异性强、快速,可作为传统RT-PCR的替代方法用于鼠疫病毒A的检测。
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