Wallis C, Melnick J L, Longoria C J
J Clin Microbiol. 1981 Sep;14(3):342-6. doi: 10.1128/jcm.14.3.342-346.1981.
An inexpensive, rapid, and simple colorimetric test for detection of bacteriuria is described. This test does not require bacterial growth and has the marked advantage of being able to quantify bacteria, even when the organisms are present in the urine of bacteriuric patients who are being treated with antibiotics. The test is carried out with 1 ml of urine, which is processed through a 10-mm-diameter filter than entraps the bacteria on its surface. Safranine dye is passed through the filter to stain the bacteria and the filter fibers. A decolorizer, which removes the dye from the filter fibers but not from the bacteria, is then passed through the filter. If there are greater than or equal to 10(5) colony-forming units of bacteria per ml in the sample, the 10-mm filter disk manifests a pink to red color. If there are less than 10(5)colony-forming units of bacteria per ml, the filter disk remains white or becomes slightly yellow. The entire procedure has been adapted to a semiautomated instrument and the time required per test is less than 1 min. The results obtained on the test card are a permanent record to be filled with the patient's chart. The bacteria can be quickly classified as gram positive or gram negative by selective staining of a second milliliter of urine on the filter. Of 441 urine specimens tested, 430 (98%) were correctly classified as containing more or less than 10(5) colony-forming units per ml. A total of 62 urine specimens were positive by bacterial plating (greater than or equal to 10(5) colony-forming units per ml), and 59 were positive by the colorimetric test. Eight false-positives (colorimetric test positive, plate counts less than 10(5)) were encountered in patients (bacteriuric) being prescribed antibiotics. Removal of the antibiotics from these urine specimens, with subsequent replating of the samples, indicated the presence of greater than or equal to 10(5) colony-forming units of bacteria per ml in three representative cases tested, indicating that the results of the colorimetric tests were not false-positives but that the plate counts were low because of the inhibition of bacterial growth by the residual antibiotics present in the urine.
本文描述了一种用于检测菌尿症的廉价、快速且简单的比色试验。该试验无需细菌生长,即使在接受抗生素治疗的菌尿症患者尿液中存在细菌时,也具有能够对细菌进行定量的显著优势。试验使用1毫升尿液,通过直径为10毫米的过滤器进行处理,细菌会截留在其表面。番红染料通过过滤器对细菌和滤膜纤维进行染色。然后通过过滤器加入一种脱色剂,该脱色剂可去除滤膜纤维上的染料,但不会去除细菌上的染料。如果样品中每毫升细菌的菌落形成单位大于或等于10⁵,则10毫米的滤膜圆盘会呈现粉红色至红色。如果每毫升细菌的菌落形成单位少于10⁵,滤膜圆盘则保持白色或变为浅黄色。整个过程已适配到半自动仪器,每次测试所需时间少于1分钟。试验卡上获得的结果是要填写在患者病历中的永久记录。通过在滤膜上对另一毫升尿液进行选择性染色,可快速将细菌分类为革兰氏阳性或革兰氏阴性。在测试的441份尿液样本中,430份(98%)被正确分类为每毫升含有或多或少10⁵个菌落形成单位。通过细菌平板培养,共有62份尿液样本呈阳性(每毫升大于或等于10⁵个菌落形成单位),通过比色试验有59份呈阳性。在接受抗生素治疗的(菌尿症)患者中出现了8例假阳性(比色试验阳性,平板计数少于10⁵)。从这些尿液样本中去除抗生素,随后对样本重新进行平板培养,结果表明在测试的三个代表性病例中,每毫升存在大于或等于10⁵个菌落形成单位的细菌,这表明比色试验结果并非假阳性,而是由于尿液中残留的抗生素抑制了细菌生长,导致平板计数较低。