Thore A, Anséhn S, Lundin A, Bergman S
J Clin Microbiol. 1975 Jan;1(1):1-8. doi: 10.1128/jcm.1.1.1-8.1975.
A selective method for distinguishing bacterial and nonbacterial adenosine triphosphate (ATP) in clinical bacteriological specimens was studied. The method involved incubation of samples with the detergent Triton X-100 and the ATP-hydrolyzing enzyme apyrase. The incubation selectively destroyed ATP in suspensions of various human cells while not affecting the ATP content in microbial cells. ATP remaining in the sample after incubation was extracted in boiling buffer and assayed by the firefly luciferase assay. Application of the method to 469 clinical urine specimens showed that the ATP level after treatment with Triton/apyrase was correlated to bacterial counts and that the sensitivity of the assay was sufficient for the detection of 10(5) bacteria/ml. The ATP levels per bacterial cell remaining in the urine specimen after treatment with Triton/apyrase were close to values observed in laboratory-grown cultures. The specificity and sensitivity of the luciferase assay for the detection of urinary bacteria and its possible use as a bacteriuria screening method are discussed.
研究了一种用于区分临床细菌学标本中细菌和非细菌三磷酸腺苷（ATP）的选择性方法。该方法包括将样品与去污剂 Triton X - 100 和 ATP 水解酶腺苷三磷酸双磷酸酶一起孵育。孵育可选择性地破坏各种人体细胞悬液中的 ATP，而不影响微生物细胞中的 ATP 含量。孵育后样品中剩余的 ATP 在沸腾缓冲液中提取，并用萤火虫荧光素酶测定法进行测定。将该方法应用于 469 份临床尿液标本，结果表明，用 Triton/腺苷三磷酸双磷酸酶处理后的 ATP 水平与细菌计数相关，并且该测定法的灵敏度足以检测到每毫升 10⁵ 个细菌。用 Triton/腺苷三磷酸双磷酸酶处理后尿液标本中每个细菌细胞的 ATP 水平接近在实验室培养物中观察到的值。讨论了荧光素酶测定法检测尿细菌的特异性和灵敏度及其作为菌尿筛查方法的可能用途。
