通过信息收集规则以及集成细胞培养-巢式聚合酶链反应程序，对采集并评估的地表水中星状病毒、肠道病毒以及40型和41型腺病毒进行检测。
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.
作者信息
Chapron C D, Ballester N A, Fontaine J H, Frades C N, Margolin A B
机构信息
Department of Microbiology, University of New Hampshire, Durham, New Hampshire 03824, USA.
出版信息
Appl Environ Microbiol. 2000 Jun;66(6):2520-5. doi: 10.1128/AEM.66.6.2520-2525.2000.
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.
我们评估了一种结合巢式PCR的细胞培养-逆转录PCR（ICC-RT-PCR）方法，用于检测通过信息收集规则（ICR）方法采集和评估的地表水样中的人星状病毒、肠道病毒以及40型和41型腺病毒。将ICC-RT-PCR-巢式PCR方法获得的结果与总可培养病毒测定-最可能数（TCVA-MPN）方法（美国环境保护局推荐的用于监测地表水和成品水中病毒的方法）获得的结果进行比较。对29个ICR地表水样本进行了分析。通过过滤吸附-牛肉提取物洗脱和有机絮凝技术浓缩病毒，然后通过观察水牛绿猴肾（BGMK）细胞系中的细胞病变效应来评估制剂中的病毒。在ICC-RT-PCR-巢式PCR技术中，我们使用Caco-２细胞增殖星状病毒和肠道病毒（ICC步骤），并使用BGMK细胞增殖40型和41型腺病毒以及肠道病毒。通过ICC-RT-PCR-巢式PCR方法测定，29个样本中有15个（51.7%）星状病毒呈阳性，其中8个样本（27.5%）含有传染性星状病毒。当使用BGMK细胞系时，29个样本中有17个（58.6%）肠道病毒呈阳性，其中6个样本（27.6%）被确定具有传染性。29个样本中有14个（48.3%）40型和41型腺病毒呈阳性，其中11个样本（37.9%）被确定具有传染性。29个样本中有27个（93.1%）病毒呈阳性，19个样本（68.9%）含有传染性病毒。通过TCVA-MPN方法测定，29个样本中只有5个（17.2%）呈阳性。与TCVA-MPN方法相比，ICC-RT-PCR-巢式PCR方法具有更高的灵敏度。
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