Detection of enteroviruses, hepatitis A virus and rotaviruses in sewage by means of an immunomagnetic capture reverse transcription-PCR assay.
Microbiol Res
; 157(3): 169-75, 2002.
Article
em En
| MEDLINE
| ID: mdl-12398285
ABSTRACT
An immunomagnetic capture reverse transcription-PCR (IMC-RT-PCR) assay was evaluated to recover and detect enteric viruses in sewage and to remove PCR inhibitors. The procedure was applied along with a simple sample processing consisting of an initial separation of solids followed by polyethylen glycol precipitation and solvent extraction. This procedure reduced sample volumes by about 65-fold without eliminating RT-PCR inhibitors. Paramagnetic beads coupled to pooled human immunoglobulins were used to simultaneously capture poliovirus 1 (PV-1) and hepatitis A virus (HAV) from seeded sewage concentrates. The IMC was efficient in removing PCR inhibitors and in further reducing sample volumes by approximately 10-fold allowing the analysis of 6-7 ml of sewage sample per RT-PCR reaction. The detection limits of IMC-RT-PCR from seeded concentrates were 0.1-1 PFU for PV-1 and 1 MPNCU for HAV. The described procedure could be applied successfully for the detection of enteroviruses, HAV and rotaviruses in field sewage samples.
Buscar no Google
Base de dados:
MEDLINE
Assunto principal:
Vírus de RNA
/
Esgotos
Tipo de estudo:
Diagnostic_studies
Idioma:
En
Revista:
Microbiol Res
Assunto da revista:
MICROBIOLOGIA
/
SAUDE AMBIENTAL
Ano de publicação:
2002
Tipo de documento:
Article