Gantzer C, Senouci S, Maul A, Levi Y, Schwartzbrod L
Laboratoire de Virologie, Faculté de Pharmacie, Nancy, France.
J Virol Methods. 1997 May;65(2):265-71. doi: 10.1016/s0166-0934(97)02193-9.
Standard methods for detecting enteroviruses in environmental samples require cell culture, which is time consuming and expensive. The reverse transcription-polymerase chain reaction (RT-PCR) is a rapid, sensitive method for detecting enteroviruses in water. However, environmental samples often contain substances that inhibit PCR amplification of target RNA. Hence the virus must be concentrated by procedures that do not interfere with amplification. This study shows that virus concentration by adsorption onto glass powder or glass wool supports is suitable for detecting viral genomes in treated wastewater by RT semi-nested PCR. No enterovirus genome was detected directly in 25 samples of treated wastewater by RT semi-nested PCR. However, samples concentrated by adsorption onto glass wool or glass powder showed that 48% (glass powder) and 56% (glass wool) contained virus. Secondary concentration by organic flocculation was unsuitable for detecting virus concentrated on glass wool (20% positive samples), but it helped to increase the detection of the genome after concentration on glass powder (72% positive samples).
检测环境样本中肠道病毒的标准方法需要细胞培养，这种方法既耗时又昂贵。逆转录-聚合酶链反应（RT-PCR）是一种快速、灵敏的检测水中肠道病毒的方法。然而，环境样本中常常含有抑制目标RNA进行PCR扩增的物质。因此，必须通过不干扰扩增的程序来浓缩病毒。本研究表明，通过吸附到玻璃粉或玻璃棉载体上进行病毒浓缩，适用于通过RT半巢式PCR检测处理后废水中的病毒基因组。通过RT半巢式PCR直接检测25个处理后废水样本时，未检测到肠道病毒基因组。然而，通过吸附到玻璃棉或玻璃粉上进行浓缩的样本显示，48%（玻璃粉）和56%（玻璃棉）的样本含有病毒。通过有机絮凝进行二次浓缩不适用于检测浓缩在玻璃棉上的病毒（阳性样本占20%），但有助于提高在玻璃粉上浓缩后病毒基因组的检测率（阳性样本占72%）。
