用于检测产毒白喉棒状杆菌的反向被动乳胶凝集试验。
Reversed passive latex agglutination assay for detection of toxigenic Corynebacterium diphtheriae.
作者信息
Toma C, Sisavath L, Iwanaga M
机构信息
Department of Bacteriology, Faculty of Medicine, University of the Ryukyus, Okinawa, Japan.
出版信息
J Clin Microbiol. 1997 Dec;35(12):3147-9. doi: 10.1128/jcm.35.12.3147-3149.1997.
A reversed passive latex agglutination (RPLA) assay for determining the toxigenicity of Corynebacterium diphtheriae is presented. Rabbit antitoxin antiserum was raised by using commercially available diphtheria toxoid. This antiserum reacted with the diphtheria toxin when the culture supernatant was assayed by Western blotting, and it did not cross-react with other extracellular antigens. Affinity-purified antibodies for latex sensitization were obtained by using a Hi Trap N-hydroxysuccinimide-activated column. Demonstration of toxin in five of seven clinical isolates was in accordance with the PCR assay and the Vero cell cytotoxicity test. Culture of the bacteria for 6 h was sufficient for toxin production, and an additional 6 h was needed to observe latex agglutination. Therefore, diphtheria toxin can be detected in 12 h by this method. The lowest concentration of diphtheria toxin detectable by the RPLA assay was about 5 ng/ml. The RPLA assay can provide a convenient and reliable method for laboratories involved in the identification of toxinogenic corynebacteria.
本文介绍了一种用于测定白喉棒状杆菌产毒性的反向被动乳胶凝集（RPLA）试验。使用市售白喉类毒素制备兔抗毒素抗血清。当通过蛋白质印迹法检测培养上清液时，该抗血清与白喉毒素发生反应，且不与其他细胞外抗原发生交叉反应。通过使用Hi Trap N-羟基琥珀酰亚胺活化柱获得用于乳胶致敏的亲和纯化抗体。七株临床分离株中有五株的毒素检测结果与聚合酶链反应（PCR）检测和Vero细胞细胞毒性试验结果一致。细菌培养6小时足以产生毒素，还需要额外6小时来观察乳胶凝集。因此，用该方法可在12小时内检测到白喉毒素。RPLA试验可检测到的白喉毒素最低浓度约为5 ng/ml。RPLA试验可为参与产毒素棒状杆菌鉴定的实验室提供一种便捷可靠的方法。
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