Identification of Klebsiella Pneumoniae by Capsular Polysaccharide Polyclonal Antibodies
Identification of Klebsiella Pneumoniae by Capsular Polysaccharide Polyclonal Antibodies
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荚膜多糖多克隆抗体鉴定肺炎克雷伯菌
DOI：
10.7763/ijcea.2011.v2.89
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发表时间：
2011
期刊：
影响因子：
--
通讯作者：
H. Batra
中科院分区：
文献类型：
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作者：
A. Sikarwar;H. Batra
本研究的目的是建立一种简便、快速的肺炎克雷伯菌的诊断方法。这项研究是在印度瓜廖尔的DRDE进行的。我们收集了来自日本山口大学的标准肺炎克雷伯菌菌株3296和来自浦那武装部队医学院和新德里帕特尔胸科医院的共59株克雷伯菌属临床分离株。将细菌生长在胰蛋白酶解大豆肉汤上，并用纯化的肺炎克雷伯氏菌荚膜多糖（CPS）皮下免疫两只新西兰Wistar白色家兔。用逆流免疫电泳法检测CPS高免血清的特异性。我们制备了四株单克隆抗体，其中两株产生IgM的克隆（KP-1和KP-2）和抗CPS抗原的高免血清用于凝集试验。KP-1、KP-2和CPS高免血清反应不同。在20株经生化确认的肺炎克雷伯氏菌临床分离株中，CPS超免疫血清检测到16株呈阳性，而KP-1对其中11株产生反应，KP-2对其中13株产生反应。通过对所有三种抗体试剂进行凝集试验，一种临床克雷伯菌属的肺炎克雷伯菌和其他肠杆菌科微生物生化阴性。然而，产气克雷伯菌对KP-2和CPS超免疫血清有反应。标准肺炎克雷伯菌对所有三种抗体均呈阳性。快速和容易的性能值得CPS和IgM单克隆抗体为基础的凝集试验作为肺炎克雷伯菌的初步鉴定系统的超免疫血清。
The aim of this study is to develop simple and rapid diagnostic method which can be utilized routinely for detection of Klebsiella pneumoniae. This study was done in India at DRDE, Gwalior. We have collected standard K.pneumoniae strain 3296 from Yamaguchi University, Japan and total fifty-nine clinical isolates of Klebsiella species from Armed Force Medical College, Pune, and Patel chest hospital, New Delhi. Bacteria were grown on trypticase soy broth and two Newzealand wistar white rabbit were immunized subcutaneously with purified capsular polysaccharide (CPS) of K.pneumoniae. Specificity of hyper immune sera raised against CPS was tested by counter current immunoelectrophoresis. We have generated four monoclonal antibodies, out of which two IgM producing clones (KP-1 & KP-2) and hyper immune sera against CPS antigen were used for agglutination test. KP-1, KP-2 and CPS hyperimmune sera showed different reactions. Out of the twenty biochemically confirmed K.pneumoniae clinical isolates, hyperimmune sera to CPS detected sixteen as positive whereas KP-1 reacted to eleven and KP-2 to thirteen of these isolates. One clinical Klebsiella species biochemically negative for K.pneumoniae and other Enterobacteriaceae organisms were negative by agglutination test to all the three antibody reagents. K.aerogenes, however, reacted to KP-2 and CPS hyperimmune sera. Standard K.pneumoniae was positive to all the three antibodies. The rapidity and ease of performance merited the hyper immune sera to CPS and IgM monoclonal antibody based agglutination test as the preliminary identification system for K.pneumoniae.