evaluation of antibodies produced against lipopolysaccharide of Vibrio cholerae for rapid detection of cholera
Abstract:
Background and Objective
Iran is one of the endemic foci of cholera and early detection, especially in times of disease outbreaks, is of vital importance. Antibodies against the components of LPS are one of the important ways for bacterial detection. The purpose of this study was to Extraction and purification of lipopolysaccharide of Vibrio cholerae, and evaluation of antibodies produced against it for detection of cholera
Materials and Methods
Vibrio cholerae was cultured in TCBS. LPS was extracted by hot phenol water method, purified and dialyzed. The content of protein and sugar, purity, and its biological activity was measured. To produce antibodies, the killed bacteria were injected into the rabbits and then over three times, LPS was injected with Freund's incomplete Adjuvant. Before and after the booster injection, blood samples were taken and the amount of antibodies against the serotypes Inaba and Ogawa, the LPS of serotypes Inaba and Ogawa, and several other similar bacteria were determined by ELISA technique.
Results
The amount of protein in purified LPS was about zero, sugar was 0.5 mg/ml, titer activity 1024, and has three 5.2, 4 and 5.14 KD bands. Antibodies produced in rabbits were able to identify the bacterial Inaba and Oagwa serotype, and also pure LPS. Ogawa and Inaba serotypes cross react against each other but not with other species of Vibrio and also other bacteria. LPS antibody titer against serotype Ogawa and serotype Inaba were 1:32000 and 1:16000, respectively.
Conclusion
Due to the low cost production and high sensitivity, and the importance of diagnosis of cholera in Iran, the antiserum produced in this study can be used as a tool for early screening of cholera and discrimination of O1 strains from non O1 as in Immunologic tests.
Language:
Persian
Published:
Pathobiology Reaearch, Volume:18 Issue: 4, 2016
Pages:
25 - 31
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