Detection of naturally occurring enteroviruses in waters by reverse transcription, polymerase chain reaction, and hybridization.
Comparison in virus-seeded mineral water of three detection methods for enteroviruses, direct hybridization, cell culture, and reverse transcription into cDNA followed by polymerase chain reaction and hybridization, showed that the last procedure was 10 to 1,000 times more sensitive than detection by cell culture and 10(5) to 10(7) times more sensitive than direct hybridization. The presence of naturally occurring enteroviruses was also demonstrated in activated sludge and in concentrated and non-concentrated surface water samples by reverse transcription-polymerase chain reaction-hybridization. However, in activated sludge and in concentrated surface waters, enzymatic amplification was sometimes inhibited by contaminants.
Kopecka
H
H
Institut für Wasser, D-1000 Berlin 33, Germany.
Dubrou
S
S
Prevot
J
J
Marechal
J
J
López-Pila
J M
JM
D000818:Animals; D001483:Base Sequence; D002522:Chlorocebus aethiops; D004277:DNA, Single-Stranded; D004770:Enterovirus / Q000254:growth & development / Q000302:isolation & purification*; D006367:HeLa Cells; D006801:Humans; D008969:Molecular Sequence Data; D009693:Nucleic Acid Hybridization; D016133:Polymerase Chain Reaction; D012367:RNA, Viral / Q000032:analysis; D012194:RNA-Directed DNA Polymerase / Q000378:metabolism; D012680:Sensitivity and Specificity; D012722:Sewage; D014709:Vero Cells; D014773:Virology / Q000379:methods*; D014776:Virus Cultivation; D014871:Water Microbiology
D004277:DNA, Single-Stranded; D012367:RNA, Viral; D012722:Sewage; D012194:RNA-Directed DNA Polymerase