Enterovirus genomes in wastewater: concentration on glass wool and glass powder and detection by RT-PCR.
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).
Gantzer
C
C
Laboratoire de Virologie, Faculté de Pharmacie, Nancy, France.
Senouci
S
S
Maul
A
A
Levi
Y
Y
Schwartzbrod
L
L
D002460:Cell Line; D004770:Enterovirus / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification*; D016679:Genome, Viral; D005898:Glass; D016133:Polymerase Chain Reaction; D011208:Powders / Q000032:analysis*; D011485:Protein Binding; D012722:Sewage / Q000821:virology*; D012996:Solutions; D014876:Water Pollution / Q000009:adverse effects / Q000032:analysis*
D011208:Powders; D012722:Sewage; D012996:Solutions