Ryan R W
Diagn Microbiol Infect Dis. 1986 Mar;4(3 Suppl):79S-86S. doi: 10.1016/s0732-8893(86)80045-1.
Clostridium difficile has been shown to be the major cause of antibiotic-associated gastroenteritis in both humans and experimental animals. During the past few years an increasing number of laboratories have attempted to detect, isolate, and identify this organism and its toxin from clinical samples. Direct visualization of C. difficile in patient specimens using immunofluorescent microscopy has been proposed. The major disadvantage of this method was its lack of specificity due to crossreaction with other clostridial species. Attempts to absorb the antisera with crossreacting strains also failed. Laboratory diagnosis of C. difficile in clinical specimens has relied on either culture using one or more selective media or on the detection of specific cytotoxin in stool filtrates. Until recently the cytotoxicity assay was the only procedure available for the routine detection of cytotoxin and, as a result, has limited this test to laboratories with access to tissue culture facilities. As a result, there has been much interest in the development of immunochemical methods for the detection of C. difficile toxins. We originally reported on the detection of C. difficile toxin in stool filtrates using counterimmunoelectrophoresis. We examined 140 fecal specimens submitted for C. difficile toxin assay by counterimmunoelectrophoresis, using both unabsorbed and absorbed antitoxin, tissue culture, and bacterial culture. Using tissue culture assay as the reference method, the sensitivity of counterimmunoelectrophoresis and counterimmunoelectrophoresis-absorbed was 100% and the specificity 63.0% and 77.5%, respectively. Enzyme immunosorbent assays for the detection of toxin A from C. difficile have also been reported, however, at the present time they do not appear to be as sensitive as the cytotoxicity assay for toxin B (cytotoxin).(ABSTRACT TRUNCATED AT 250 WORDS)
艰难梭菌已被证明是人类和实验动物抗生素相关性肠胃炎的主要病因。在过去几年中,越来越多的实验室试图从临床样本中检测、分离和鉴定这种微生物及其毒素。有人提出使用免疫荧光显微镜直接观察患者标本中的艰难梭菌。该方法的主要缺点是由于与其他梭菌属物种发生交叉反应而缺乏特异性。用交叉反应菌株吸收抗血清的尝试也失败了。临床标本中艰难梭菌的实验室诊断依赖于使用一种或多种选择性培养基进行培养,或检测粪便滤液中的特定细胞毒素。直到最近,细胞毒性测定仍是常规检测细胞毒素的唯一方法,因此,该检测仅限于能够使用组织培养设施的实验室。因此,人们对开发用于检测艰难梭菌毒素的免疫化学方法非常感兴趣。我们最初报道了使用对流免疫电泳检测粪便滤液中的艰难梭菌毒素。我们使用未吸收和吸收的抗毒素、组织培养和细菌培养,通过对流免疫电泳检查了140份提交进行艰难梭菌毒素检测的粪便标本。以组织培养测定作为参考方法,对流免疫电泳和吸收后的对流免疫电泳的灵敏度分别为100%,特异性分别为63.0%和77.5%。也有报道称酶联免疫吸附测定法可检测艰难梭菌的毒素A,但目前它们似乎不如毒素B(细胞毒素)的细胞毒性测定法灵敏。(摘要截取自250字)