Johnson L L, McFarland L V, Dearing P, Raisys V, Schoenknecht F D
University Hospital, Seattle, Washington.
J Clin Microbiol. 1989 Oct;27(10):2218-21. doi: 10.1128/jcm.27.10.2218-2221.1989.
We have developed a sensitive and specific method for the identification of Clostridium difficile in stool specimens based on the detection of metabolic breakdown products of the organism by gas-liquid chromatography after incubation of stool samples in a selective broth medium containing cefoxitin. Use of this approach to test samples from two different populations of patients at separate medical centers showed this method to be superior to plate cultures or cytotoxin testing alone for both populations. The combined results from the two patient populations showed that 225 of 226 confirmed isolates were identified correctly, resulting in a sensitivity of 99.6% and a specificity of 99.0%. This method eliminates the delay caused by subculturing for tests requiring a pure isolate. The culture phase amplifies even low numbers of C. difficile in fecal samples (due to low in vivo concentrations or delayed transport) and thus increases sensitivity. Other advantages include the ability to detect C. difficile in the mixed flora of the stool and the ability of most clinical laboratories to use this procedure. Given the complexities of the detection of C. difficile toxins and the increasing importance of this organism as a nosocomial agent, culture-based methods remain the preferred approach to screening and routine workup for cases of diarrhea.
我们开发了一种灵敏且特异的方法,用于在粪便标本中鉴定艰难梭菌。该方法基于在含有头孢西丁的选择性肉汤培养基中孵育粪便样本后,通过气液色谱法检测该菌的代谢分解产物。运用此方法对来自两个不同医疗中心的不同患者群体的样本进行检测,结果表明,对于这两个群体,该方法均优于单独的平板培养法或细胞毒素检测法。两个患者群体的综合检测结果显示,226株已确认的分离株中有225株被正确鉴定,灵敏度为99.6%,特异性为99.0%。该方法消除了因需要纯分离株进行检测而进行传代培养所导致的延迟。培养阶段可扩增粪便样本中即便数量很少的艰难梭菌(由于体内浓度低或运输延迟),从而提高了灵敏度。其他优点包括能够在粪便的混合菌群中检测到艰难梭菌,以及大多数临床实验室都能够采用此方法。鉴于艰难梭菌毒素检测的复杂性以及该菌作为医院感染病原体的重要性日益增加,基于培养的方法仍然是腹泻病例筛查和常规检查的首选方法。