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首页> 美国卫生研究院文献>Applied and Environmental Microbiology >Detection of Astroviruses Enteroviruses and Adenovirus Types 40 and 41 in Surface Waters Collected and Evaluated by the Information Collection Rule and an Integrated Cell Culture-Nested PCR Procedure
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Detection of Astroviruses Enteroviruses and Adenovirus Types 40 and 41 in Surface Waters Collected and Evaluated by the Information Collection Rule and an Integrated Cell Culture-Nested PCR Procedure

机译：通过信息收集规则和集成的细胞培养巢式PCR程序收集和评估地表水中的星状病毒肠病毒和40和41型腺病毒的检测

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We evaluated the use of an integrated cell culture-reverse transcription-PCR (ICC-RT-PCR) procedure coupled with nested PCR to detect human astroviruses, enteroviruses, and adenovirus types 40 and 41 in surface water samples that were collected and evaluated by using the Information Collection Rule (ICR) method. The results obtained with the ICC-RT-PCR–nested PCR method were compared to the results obtained with the total culturable virus assay–most-probable-number (TCVA-MPN) method, the method recommended by the U.S. Environmental Protection Agency for monitoring viruses in surface and finished waters. Twenty-nine ICR surface water samples were analyzed. Viruses were concentrated by using filter adsorption-beef extract elution and organic flocculation techniques, and then the preparations were evaluated for viruses by visualizing cytopathic effects in the Buffalo green monkey kidney (BGMK) cell line. In the ICC-RT-PCR–nested PCR technique we used Caco-2 cells to propagate astroviruses and enteroviruses (ICC step), and we used BGMK cells to propagate adenovirus types 40 and 41, as well as enteroviruses. Fifteen of the 29 samples (51.7%) were positive for astrovirus as determined by the ICC-RT-PCR–nested PCR method, and eight of these samples (27.5%) contained infectious astrovirus. Seventeen of the 29 samples (58.6%) were positive for enteroviruses when the BGMK cell line was used, and six (27.6%) of these samples were determined to be infectious. Fourteen of the 29 samples (48.3%) were positive for adenovirus types 40 and 41, and 11 (37.9%) of these samples were determined to be infectious. Twenty-seven of the 29 samples (93.1%) were positive for a virus, and 19 (68.9%) of the samples were positive for an infectious virus. Only 5 of the 29 samples (17.2%) were positive as determined by the TCVA-MPN method. The ICC-RT-PCR–nested PCR method provided increased sensitivity compared to the TCVA-MPN method.
机译：我们评估了使用集成的细胞培养逆转录PCR（ICC-RT-PCR）程序和嵌套式PCR来检测地表水样品中人类星状病毒，肠病毒以及40和41型腺病毒的类型，这些样本是通过使用信息收集规则（ICR）方法。将使用ICC-RT-PCR嵌套式PCR方法获得的结果与通过总可培养病毒测定法-最有可能发生数（TCVA-MPN）方法获得的结果进行比较，该方法是美国环境保护署推荐的监测方法地表水中和成品水中的病毒。分析了29个ICR地表水样品。通过使用滤器吸附牛肉提取物洗脱和有机絮凝技术浓缩病毒，然后通过可视化布法罗绿猴肾（BGMK）细胞系的细胞病变作用评估制剂中的病毒。在ICC-RT-PCR嵌套PCR技术中，我们使用Caco-2细胞繁殖星状病毒和肠病毒（ICC步骤），并使用BGMK细胞繁殖40和41型腺病毒以及肠道病毒。根据ICC-RT-PCR巢式PCR方法测定，在29个样本中有15个（51.7％）呈星形病毒阳性，其中8个样本（27.5％）含有传染性星状病毒。使用BGMK细胞系时，这29个样品中有17个（58.6％）呈肠道病毒阳性，其中六个（27.6％）被确定具有传染性。 29个样品中有14个（48.3％）对40和41型腺病毒呈阳性，其中11个（37.9％）被确定为具有传染性。 29个样本中有27个（93.1％）对病毒呈阳性，其中19个（68.9％）对传染性病毒呈阳性。通过TCVA-MPN方法测定的29个样品中只有5个（17.2％）为阳性。与TCVA-MPN方法相比，ICC-RT-PCR嵌套PCR方法提供了更高的灵敏度。

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