Chemistry: molecular biology and microbiology – Measuring or testing process involving enzymes or... – Involving oxidoreductase
Patent
1991-06-03
1993-11-23
Wityshyn, Michael G.
Chemistry: molecular biology and microbiology
Measuring or testing process involving enzymes or...
Involving oxidoreductase
435 4, 435 18, 435 28, 435 32, 435 39, 435 47, 435 49, 435231, 435871, 435933, 435973, 436 73, 436904, 514192, 514200, C12Q 126, C12Q 128
Patent
active
052643468
ABSTRACT:
A novel colorimetric method for beta-lactamase assay and its applications are disclosed. The novel assay method for beta-lactamase is based on the discovery described in this invention. The chromophore formed by oxidation of either the N-alkyl derivative of p-phenylenediamine or the 3,3',5,5'-tetraalkyl derivative of benzidine can be decolorized by the open beta-lactam ring end product resulting from the hydrolysis of a penicillin in the presence of a mercury-containing compound. The same chromophore can be decolorized by the open beta-lactam ring end product resulting from the hydrolysis of a cephalosporin in either the presence or the absence of the mercury-containing compound. None of the intact beta-lactam antibiotics can decolorize the chromophore in either the presence or the absence of the mercury-containing compound. The final concentration of the mercury-containing compound in the decolorization mixture ranges from approximately 0.005 mM to approximately 3 mM, and the pH of the decolorization mixture ranges from 3.5 to 8.0. Additional applications derived from coupling this novel assay method to methods in the art include: a method for specific detection of microbial beta-lactamases; a method for simultaneous detection of microbial cytochrome c oxidase systems and penicillinases, and its application to simultaneous presumptive identification of Neisseria gonorrhoeae in urethral exudates, and differentiation of the infecting gonococci into penicillinase-producing N. gonorrhoeae and non-penicillinase-producing N. gonorrhoeae; and a simultaneous assay method for peroxidase and penicillinase.
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Mohamed Abdel A.
Wityshyn Michael G.
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