Pretreatment to avoid positive RT-PCR results with inactivated viruses
Pretreatment to avoid positive RT-PCR results with inactivated viruses
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DOI：
10.1016/s0166-0934(02)00089-7
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发表时间：
2002-07-01
影响因子：
3.1
通讯作者：
Cliver, DO
中科院分区：
文献类型：
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作者：
Nuanualsuwan, S;Cliver, DO
肠道病毒是人类疾病的重要原因，通常必须通过逆转录-聚合酶链式反应(RT-PCR)进行检测，这种方法通常对只含有灭活病毒的样本产生阳性结果。本研究旨在开发一种样品的前处理方法，以使RT-PCR程序不会检测到灭活病毒。模型病毒是人类甲型肝炎病毒、疫苗脊髓灰质炎病毒I和猫杯状病毒作为诺瓦克样病毒的替代品。每种病毒都被紫外线、次氯酸盐或72摄氏度加热灭活(初始滴度约为10(3)pfu/ml)。灭活病毒经37℃处理30min后，RT-PCR结果为阴性，即反应完成后未检测到扩增产物。这种RT-PCR方法的前提条件可能适用于其他类型的病毒、以其他方式灭活的病毒以及其他分子病毒检测方法。(C)2002 Elsevier Science B.V.保留所有权利。
Enteric viruses that are important causes of human disease must often be detected by reverse transcription-polymerase chain reaction (RT-PCR), a method that commonly yields positive results with samples that contain only inactivated virus. This study was intended to develop a pretreatment for samples, so that inactivated viruses would not be detected by the RT-PCR procedure. Model viruses were human hepatitis A virus, vaccine poliovirus I and feline calicivirus as a surrogate for the Norwalk-like viruses. Each virus was inactivated (from an initial titer of approximate to10(3) PFU/ml) by ultraviolet light, hypochlorite or heating at 72 degreesC. Inactivated viruses, that were treated with proteinase K and ribonuclease for 30 min at 37 degreesC before RT-PCR, gave a negative result, which is to say that no amplicon was detected after the reaction was completed. This antecedent to the RT-PCR method may be applicable to other types of viruses, to viruses inactivated in other ways and to other molecular methods of virus detection. (C) 2002 Elsevier Science B.V. All rights reserved.