Rapid parallel detection of hygienically relevant microorganisms in water samples by PCR and specific hybridization in microtiter plates.
A molecular biological test protocol for the parallel detection of enterococci and Pseudomonas aeruginosa in drinking water was developed. Amplicons labelled with digoxigenin during PCR were hybridized to specific 23S rDNA targeted oligonucleotide probes immobilized in microtiter plates. Detection was performed by addition of anti-digoxigenin-peroxidase-conjugate and chromogenic substrate. Specificity of the probes was evaluated by using pure cultures. First evaluation data with natural water samples in comparison to conventional microbiological analysis according to the German Drinking Water Regulation showed good agreement. Its feasible and rapid performance should be advantageous for use in routine drinking water quality control. Further comparative evaluation studies need to be undertaken to determine the true applicability for routine testing of water samples.
Frahm
E
E
Forschungszentrum Karlsruhe GmbH, ITC-WGT, Abt. Umweltmikrobiologie, Germany.
Heiber
I
I
Ludwig
W
W
Obst
U
U
D006920:Hygiene; D009693:Nucleic Acid Hybridization / Q000379:methods*; D016133:Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D014871:Water Microbiology; D014881:Water Supply / Q000592:standards*