Solid-phase polymerase chain reaction: applications for direct detection of enteric pathogens in waters.
The techniques in current use for detection of pathogens in environmental samples are restricted to those organisms whose replication in either culture media or cell culture is feasible. These methods lack the selectivity and sensitivity necessary for their unequivocal detection and identification. We have developed an assay for the detection of bacterial cells in large volumes of water. Low concentrations of cells containing target sequences were concentrated on membrane filters and were subjected to amplification directly using a stepwise polymerase chain reaction. This procedure, together with nucleic acid probes, has enhanced the limit of detection to the level of a single bacterial cell. This technique could be used for the detection of any bacteria or virus in water or air.
Toranzos
G A
GA
Department of Biology, College of Natural Sciences, University of Puerto Rico, Rio Piedras 00931.
Alvarez
A J
AJ
D001483:Base Sequence; D004269:DNA, Bacterial; D004755:Enterobacteriaceae / Q000235:genetics / Q000302:isolation & purification*; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods*; D014871:Water Microbiology
D004269:DNA, Bacterial