Ma J F, Straub T M, Pepper I L, Gerba C P
Department of Microbiology and Immunology, University of Arizona, Tucson 85721.
Appl Environ Microbiol. 1994 Nov;60(11):4203-6. doi: 10.1128/aem.60.11.4203-4206.1994.
Inactivation of poliovirus type 1 by 1 N HCl, 1 N NaOH, 0.5 and 1.0 mg of free chlorine per liter, and UV light was compared by using cell culture and seminested PCR (30 cycles of reverse transcriptase-PCR plus 30 cycles of seminested PCR). A minimum contact time of 45 min with HCl, 3 min with NaOH, 3 and 6 min with 1.0 and 0.5 mg of free chlorine per liter, respectively, was required to render 1.64 x 10(2) PFU of poliovirus type 1 per ml undetectable by seminested PCR. In cell culture, a minimum contact time of 5 min to HCl, 30 s to NaOH, and 1 min to either chlorine concentration was required to render the viruses undetectable by the plaque assay method. No correlation was observed between results by PCR and cell culture when viruses were exposed to UV light. These data suggest that inactivated virus with intact nucleic acid sequences can be detected by PCR.
通过细胞培养和半巢式PCR(逆转录PCR 30个循环加半巢式PCR 30个循环)比较了1N盐酸、1N氢氧化钠、每升0.5和1.0毫克游离氯以及紫外线对1型脊髓灰质炎病毒的灭活效果。要使每毫升1.64×10² PFU的1型脊髓灰质炎病毒通过半巢式PCR检测不到,分别需要与盐酸至少接触45分钟、与氢氧化钠接触3分钟、与每升1.0和0.5毫克游离氯分别接触3分钟和6分钟。在细胞培养中,要使病毒通过空斑试验方法检测不到,分别需要与盐酸至少接触5分钟、与氢氧化钠接触30秒、与任一氯浓度接触1分钟。当病毒暴露于紫外线时,PCR和细胞培养结果之间未观察到相关性。这些数据表明,具有完整核酸序列的灭活病毒可通过PCR检测到。