Rapid chemiluminescent nucleic acid assays for detection of TEM-1 beta-lactamase-mediated penicillin resistance in Neisseria gonorrhoeae and other bacteria.
Two new assays for the detection of TEM-1 beta-lactamase-mediated bacterial penicillin resistance were developed that involve the use of specific nucleic acid hybridization. Both techniques are based on a solution-phase hybridization of oligonucleotide probes to the target DNA sequence, solid-phase capture of the probe-target complex, and an amplified chemiluminescent labeling method. One configuration of hybridization probes detected the presence of TEM-1 in Neisseria gonorrhoeae (45 strains), Haemophilus spp., Escherichia coli, Shigella sonnei and Salmonella typhi. A second configuration (TEM-1NH) detected TEM-1 beta-lactamase-mediated penicillin resistance only in N. gonorrhoeae (97 strains) and Haemophilus (6 strains) isolates in which TEM-1 is inserted in a pFA7-type plasmid. Both methods were 100 times more sensitive than a commercially available colorimetric beta-lactamase activity test and approximately 5 times more sensitive than radioisotopic dot blot screening for the gene. The assays are particularly well suited to the analysis of large numbers of samples, can be performed in a total of 4 h, and are sensitive to 10(4) to 10(5) CFU.
Sanchez-Pescador
R
R
Chiron Corporation, Emeryville, California 94608.
Stempien
M S
MS
Urdea
M S
MS
D001419:Bacteria / Q000187:drug effects; D001483:Base Sequence; D004251:DNA Transposable Elements; D008163:Luminescent Measurements; D008969:Molecular Sequence Data; D009344:Neisseria gonorrhoeae / Q000187:drug effects* / Q000235:genetics; D009693:Nucleic Acid Hybridization; D010403:Penicillin Resistance; D001618:beta-Lactamases / Q000235:genetics*
D004251:DNA Transposable Elements; D001618:beta-Lactamases