CETESB State Agency for Environmental Control, Av Prof Frederico Hermann Jr 345, CEP 05489-900, Sao Paulo, Brazil.
World J Microbiol Biotechnol. 1993 Sep;9(5):566-9. doi: 10.1007/BF00386295.
A rapid and sensitive method for the detection of wild poliovirus from sewage samples using the polymerase chain reaction (PCR) technique was investigated. To eliminate the toxicity of sample concentrates to the enzymatic system used in PCR, a methodology was developed for the purification of these concentrates, consisting of treatment with trichlorofluoroethane and Sephadex column chromatography. The viral RNA was extracted from the purified concentrates, submitted to PCR with primers specific for Brazilian wild poliovirus type 1 and for Sabin types 1, 2 and 3. The amplified products were detected by electrophoresis in vertical polyacrylamide gels and stained with ethidium bromide. The results suggest that sewage sampling for environmental surveillance, combined with the rapid and precise PCR technology, provides a powerful tool for assessment of the success of the poliovirus eradication programme.
本研究旨在探索一种利用聚合酶链反应（PCR）技术快速、灵敏检测污水样本中野生脊灰病毒的方法。为了消除样品浓缩物对 PCR 酶系统的毒性，我们开发了一种浓缩物的纯化方法，包括三氯氟乙烷处理和葡聚糖凝胶柱层析。从纯化的浓缩物中提取病毒 RNA，用针对巴西野生脊灰病毒 1 型和萨宾 1、2 和 3 型的引物进行 PCR。扩增产物通过垂直聚丙烯酰胺凝胶电泳检测，并溴化乙锭染色。结果表明，结合快速、精确的 PCR 技术的污水采样进行环境监测，为评估脊灰病毒根除计划的成功提供了有力工具。
