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首页> 美国政府科技报告 >Application of Firefly Luciferase Assay for Adenosine Triphosphate (ATP) to Antimicrobial Drug Sensitivity Testing
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Application of Firefly Luciferase Assay for Adenosine Triphosphate (ATP) to Antimicrobial Drug Sensitivity Testing

机译：萤火虫荧光素酶检测三磷酸腺苷（aTp）在抗菌药物敏感性试验中的应用

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This report documents the development of a rapid method for determining microbial suscepti¬bilities to antibiotics using the firefly luciferase assay for adenosine triphosphate (ATP). The reduction of bacterial ATP by an antimicrobial agent was de tormined to be a valid measure of drug effect in most cases. A basic procedure for determining microbial sensitivity evolved in which broth cultures of log phase bacteria (106per ml) were grown for 25 hours at antimicro¬bial concentrations which resulted in the best discrimination between "sensitive" and "resistant strains. Drug effect was quantitated by comparing ATP levels before and after incubation in the presence of antibiotic with ATP levels of an antibiotic-free growth control.nThe effect of 12 antibiotics on 8 different bacterial species gave a 94% correlation with the standard Kirby-Bauer agar disc diffusion method. A 93% correlation was obtained when the ATP assay method was applied directly to 50 urine specimens from patients with urinary tract infections. Urine samples were centrifuged first so that bacterial pellets could be suspended in broth. No primary isolation or subculturing was required. Mixed cultures in which one species was predominant gave accurate results for the most abundant organism. Since the method is based on an increase in bacterial ATP with time, the presence of leukocytes did not interfere with the interpretation of results. Both the incubation procedure and the ATP assays are com¬patible with automation.nThis report documents the development of a rapid method for determining microbial suscepti¬bilities to antibiotics using the firefly luciferase assay for adenosine triphosphate (ATP). The reduction of bacterial ATP by an antimicrobial agent was de tormined to be a valid measure of drug effect in most cases. A basic procedure for determining microbial sensitivity evolved in which broth cultures of log phase bacteria (10...6 per ml) were grown for 25 hours at antimicro¬bial concentrations which resulted in the best discrimination between "sensitive" and "resistant strains. Drug effect was quantitated by comparing ATP levels before and after incubation in the presence of antibiotic with ATP levels of an antibiotic-free growth control.nThe effect of 12 antibiotics on 8 different bacterial species gave a 94% correlation with the standard Kirby-Bauer agar disc diffusion method. A 93% correlation was obtained when the ATP assay method was applied directly to 50 urine specimens from patients with urinary tract infections. Urine samples were centrifuged first so that bacterial pellets could be suspended in broth. No primary isolation or subculturing was required. Mixed cultures in which one species was predominant gave accurate results for the most abundant organism. Since the method is based on an increase in bacterial ATP with time, the presence of leukocytes did not interfere with the interpretation of results. Both the incubation procedure and the ATP assays are com¬patible with automation.

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