Cholera DFA: an improved direct fluorescent monoclonal antibody staining kit for rapid detection and enumeration of Vibrio cholerae O1.
Cholera DFA: an improved direct fluorescent monoclonal antibody staining kit for rapid detection and enumeration of Vibrio cholerae O1.
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Cholera DFA：一种改进的直接荧光单克隆抗体染色试剂盒，用于快速检测和计数霍乱弧菌 O1。
DOI：
10.1111/j.1574-6968.1994.tb07021.x
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发表时间：
1994
影响因子：
2.1
通讯作者：
R. Colwell
中科院分区：
文献类型：
--
作者：
J. A. Hasan;D. Bernstein;A. Huq;L. Loomis;M. Tamplin;R. Colwell
本文报道了一种改良的霍乱荧光单克隆抗体染色试剂盒--霍乱DFA，该试剂盒采用特异性高的抗A抗原单克隆抗体COLTA，经异硫氰酸荧光素（FITC）标记，可直接检测和计数霍乱弧菌O 1。在甘油封固剂中使用最佳量的抗光漂白剂，以延缓荧光显微镜检查期间免疫荧光染色细胞的快速褪色，从而能够延长对单个视野的检查，以及改进结果的摄影记录而不损失荧光强度。当检测特异性时，所有30株霍乱弧菌O 1菌株均与霍乱DFA反应，而检测的100种异源菌种均不反应，因此本研究中检测的所有菌株均具有100%的特异性。在孟加拉国进行了一项现场试验，采用霍乱DFA和结果进行了比较，通过传统的培养方法。在检测的44份腹泻粪便标本中，所有培养阳性粪便标本中霍乱DFA对霍乱弧菌O 1呈阳性，所有培养阴性粪便标本中霍乱DFA对霍乱弧菌O 1呈阴性。该方法是敏感的和高度特异性的，以及简单的，即，比间接荧光分析法简单，仅需要一种试剂，在30分钟内完成染色过程，同时延缓荧光显微镜经常发生的快速褪色。
An improved fluorescent monoclonal antibody staining kit, Cholera DFA, for direct detection and enumeration of Vibrio cholerae O1 has been developed, employing a highly specific anti-A antigen monoclonal antibody, COLTA, labeled with fluorescein isothiocyanate (FITC). An optimized quantity of anti-photobleaching agent is used in a glycerol mounting medium to retard the rapid fading of immunofluorescent stained cells during fluorescent microscopy, thus enabling prolonged inspection of individual fields, as well as improved photographic recording of results without loss of fluorescence intensity. When tested for specificity, all 30 strains of V. cholerae O1 reacted with Cholera DFA, whereas 100 heterologous species examined did not, yielding 100% specificity for all strains examined in this study. A field trial was conducted in Bangladesh, employing Cholera DFA and the results were compared with those obtained by conventional culture methods. Of 44 diarrheal stool specimens tested, Cholera DFA was positive for V. cholerae O1 in all culture-positive stool specimens and negative for all culture-negative stool specimens. The procedure is sensitive and highly specific, as well as simple, i.e., less complex than the indirect fluorescent assay, requiring only one reagent and less than 30 min to complete the staining process, while retarding rapid fading that often occurs with fluorescent microscopy.