Thore A, Lundin A, Anséhn S
J Clin Microbiol. 1983 Feb;17(2):218-24. doi: 10.1128/jcm.17.2.218-224.1983.
A rapid (15 min) test for bacteriuria based on firefly luciferase analysis of bacterial ATP has been evaluated in 2,018 clinical urine specimens. The test procedure involves removal of nonbacterial ATP by treatment of urine with Triton X-100 and apyrase, extraction of bacterial ATP by boiling, and bioluminescent analysis of bacterial ATP by firefly luciferase, using a luminometer. For comparison, the widely used nitrite test was included in the study as an example of an alternative rapid chemical test. The test was set up to distinguish between specimens yielding greater than 10(5) CFU/ml and specimens yielding less than 10(5) CFU/ml. A level of 13.5 nM ATP was chosen to define the limit between negative and positive results. At this discriminatory level, 92% of specimens yielding greater than 10(5) CFU/ml and 88% of specimens yielding less than 10(5) CFU/ml were correctly classified with the luciferase method, whereas corresponding figures for the nitrite test were 55 and 99%, respectively. Of the 12% false luciferase positives, 20% were shown to contain greater than 10(5) CFU/ml on prolonged incubation, thus reducing the false-positive rate to 10%. Of the 8% false luciferase negatives, 65% had low levels of CFU in the range of 10(5) to 10(6).
一种基于萤火虫荧光素酶分析细菌ATP的快速（15分钟）菌尿检测方法已在2018份临床尿液标本中进行了评估。检测程序包括用 Triton X - 100和腺苷三磷酸双磷酸酶处理尿液以去除非细菌ATP，通过煮沸提取细菌ATP，并用荧光计通过萤火虫荧光素酶对细菌ATP进行生物发光分析。作为另一种快速化学检测的示例，该研究纳入了广泛使用的亚硝酸盐检测以作比较。该检测旨在区分每毫升产生大于10⁵CFU的标本和每毫升产生小于10⁵CFU的标本。选择13.5 nM ATP的水平来定义阴性和阳性结果之间的界限。在此鉴别水平下，荧光素酶法对每毫升产生大于10⁵CFU的标本正确分类率为92%，对每毫升产生小于10⁵CFU的标本正确分类率为88%，而亚硝酸盐检测的相应数字分别为55%和99%。在12%的荧光素酶假阳性中，20%在延长培养后显示每毫升含有大于10⁵CFU，从而将假阳性率降低至10%。在8%的荧光素酶假阴性中，65%的CFU水平较低，在10⁵至10⁶范围内。
