APPLICATION OF THE POLYMERASE CHAIN-REACTION (PCR) TO POLIOMYELITIS SURVEILLANCE THROUGH THE ANALYSES OF SEWAGE SAMPLES

A rapid and sensitive method for the detection of wild poliovirus from sewage samples using the polymerase chain reaction (PCR) technique wa s investigated. To eliminate the toxicity of sample concentrates to th e enzymatic system used in PCR, a methodology was developed for the pu rification of these concentrates, consisting of treatment with trichlo rofluoroethane and Sephadex column chromatography. The viral RNA was e xtracted 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 ve rtical polyacrylamide gels and stained with ethidium bromide. The resu lts suggest that sewage sampling for environmental surveillance, combi ned with the rapid and precise PCR technology, provides a powerful too l for assessment of the success of the poliovirus eradication programm e.