Nucleic acid extraction from polluted estuarine water for detection of viruses and bacteria by PCR and RT-PCR analysis.
We describe an extraction protocol for genomic DNA and RNA of both viruses and bacteria from polluted estuary water. This procedure was adapted to the molecular study of microflora of estuarine water where bacteria and viruses are found free, forming low-density biofilms, or intimately associated with organo-mineral particles. The sensitivity of the method was determined with seeded samples for RT-PCR and PCR analysis of viruses (10 virions/mL), and bacteria (1 colony-forming unit mL). We report an example of molecular detection of both poliovirus and Salmonella in the Seine estuary (France) and an approach to studying their association with organo-mineral particles.
Petit
F
F
Laboratoire de microbiologie du froid, UPRES 2123, IFRS 61 CNRS/Inserm, Faculté des sciences, Mont-Saint-Aignan, France. fabienne@univ-nover.fr
Craquelin
S
S
Guespin-Michel
J
J
Buffet-Janvresse
C
C
D015415:Biomarkers; D009696:Nucleic Acids / Q000032:analysis*; D017955:Poliovirus / Q000235:genetics / Q000302:isolation & purification*; D016133:Polymerase Chain Reaction / Q000379:methods*; D012475:Salmonella / Q000235:genetics / Q000302:isolation & purification*; D014871:Water Microbiology; D014876:Water Pollution / Q000032:analysis*
D015415:Biomarkers; D009696:Nucleic Acids