Identification of Klebsiella Pneumoniae by Capsular Polysaccharide Polyclonal Antibodies

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.