Enhancement of fluorescence development of end products by use of a fluorescence developer solution in a rapid and sensitive fluorescent spot test for specific detection of microbial beta-lactamases.
A fluorescent spot test method for specific detection of microbial beta-lactamases as previously published (K. C. S. Chen, J. S. Knapp, and K. K. Holmes, J. Clin. Microbiol. 19:818-825, 1984) was improved by the use of a fluorescence developer solution. The fluorescence developer solution used in this study consisted of 0.78 M sodium tartrate buffer containing 12% formaldehyde at a final pH of 4.5. An addition of 1 volume of fluorescence developer solution to 5 volumes of ampicillin or cephalex substrate solution incubated with beta-lactamase-producing organisms, followed by heating the mixture at 45 degrees C for 10 min resulted in enhancement of fluorescence of the end products of beta-lactamase activity. This provides a more sensitive assay for microbial beta-lactamases and offers the potential for direct detection of beta-lactamases in clinical specimens.
Chen
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Holmes
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KK
D000581:Amidohydrolases / Q000378:metabolism; D000658:Amoxicillin / Q000378:metabolism; D000667:Ampicillin / Q000378:metabolism*; D001419:Bacteria / Q000201:enzymology*; D002434:Cefadroxil / Q000378:metabolism; D002506:Cephalexin / Q000378:metabolism*; D002507:Cephaloglycin / Q000378:metabolism; D002510:Cephalosporinase / Q000378:metabolism; D004754:Enterobacter / Q000201:enzymology; D005453:Fluorescence; D006193:Haemophilus influenzae / Q000201:enzymology; D006358:Hot Temperature; D009344:Neisseria gonorrhoeae / Q000201:enzymology; D010405:Penicillinase / Q000378:metabolism; D013211:Staphylococcus aureus / Q000201:enzymology; D001618:beta-Lactamases / Q000032:analysis* / Q000378:metabolism
D002434:Cefadroxil; D000667:Ampicillin; D000658:Amoxicillin; D000581:Amidohydrolases; C019665:amidase; D002510:Cephalosporinase; D010405:Penicillinase; D001618:beta-Lactamases; D002507:Cephaloglycin; D002506:Cephalexin