Rapid detection of waterborne viruses using the polymerase chain reaction and a gene probe.
We describe a membrane-filter-based urea-arginine phosphate buffer method for concentrating waterborne viruses from large volumes of water to microlitre volumes, and their subsequent detection by the polymerase chain reaction (PCR). The detection step involves the extraction of RNA, synthesis of complementary DNA, amplification by PCR of target DNA with specific primers, and confirmation through nucleic acid hybridization with a radiolabelled oligonucleotide probe. The PCR technique detected the presence of enteroviruses in spiked as well as in contaminated water samples. The technique is sensitive and detects as few as 120 waterborne viral particles. PCR is simple, rapid, sensitive, specific and adaptable for water quality surveillance in less developed countries.
Jothikumar
N
N
National Environmental Engineering Research Institute, Nagpur, India.
Khanna
P
P
Kamatchiammal
S
S
Murugan
R P
RP
D001483:Base Sequence; D015139:Blotting, Southern; D003101:Collodion; D005374:Filtration / Q000379:methods; D005814:Genes, Viral / Q000235:genetics; D008567:Membranes, Artificial; D008969:Molecular Sequence Data; D017955:Poliovirus / Q000235:genetics / Q000302:isolation & purification*; D016133:Polymerase Chain Reaction / Q000379:methods*; D012367:RNA, Viral / Q000302:isolation & purification; D012680:Sensitivity and Specificity; D017361:Viral Nonstructural Proteins / Q000235:genetics; D014871:Water Microbiology
D008567:Membranes, Artificial; D012367:RNA, Viral; D017361:Viral Nonstructural Proteins; D003101:Collodion