Patterns of sonic extract of pseudomonas pseudomallei from clinical isolates

Abstract

The purposes of this study were to examine the molecular weight pattern of sonic extract of clinical isolated P. pseudomallei by SDS-PAGE and also possible antigenic relationship between this organism to other bacteria by an immunoblot assay. The sonic extracts of 32 clinically isolated strains of P. pseudomallei were subjected to the SDS-PAGE technique, and were able to differentiate into six types. In addition to the intensity of the bands, the presence of bands at molecular weights of 56.0, 27.5, 19.9, 19.6, 14.5, 14.4 and 12.6 Kd were used as the criteria for this typing. The 32 strains could be typed as follows: 9 strains were in Type I;6 strains were in Type II; 1 strain was in Type III; 3 strains were in Type IV; 3 strains were in Type V and 10 strains were in Type VI. A significant correlation between the anatomic sites of infection, geographic locations, and SDS-PAGE types were not found. These clinically isolated strains were further studied by the immunoblot technique using, hyperimmune antiserum against P.pseudomallei NCTC 4845. The visible antigenic band allowed the classification of our 32 isolates into three groups: A, B and C. The correlation between the SDS-PAGE typing and the immunoblot grouping was noted. SDS-PAGE Type I and II were within Immunoblot group A, Type IV was within group B. and the Type III, V and VI were in group C. In addition, other organisms such as P. aeruginosa ATCC 27853, P. cepacia JCM 5510 , P. stutzeri JCM 5969, P. putida JCM 6160, P. multophila JCM 3801, V.cholerae 569B, S. typhi NCTC 781, E. coli ATCC 25922 and S. aureus ATCC 25923 showed antigenic bands ranging from 12.0 to 140.0 Kd which were commonly found in all of the P. pseudomallei isolates. Interestingly antigenic bands at the molelular weight of 16.8, 20.7, 24.1, 107.0, 115.0, and 140.0 which were in common to all strains of P. pseudomallei were absent from the bacterial strains named above. Result obtained from this study may be useful as basic knowledge in the further investigation for common specific epitope of P. pseudomallei in the application for production of diagnostic kit as well as vaccine production. In addition they can be use for study of basic sciences, taxonomy, epidemiology, and serology of this organisms.