Application of the polymerase chain reaction (PCR) to poliomyelitis surveillance through the analyses of sewage samples

Abstract

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.