USE OF VARIOUS METHODS TO ISOLATEEXOPOLYSACCHARIDE OF ELTOR CHOLERA VIBRIOS AND ITS IMMUNOCHEMICALCHARACTERISTICS
- Authors: Tatarenko OA1, Alekseeva LP1, Telesmanich NR1, Markina OV1, Chemisova OS1, Pisanov RV1, Kruglikov VD1, Shestialtynova IS1, Uskova NN1, Tatarenko OA2, Alekseeva LP2, Telesmanich NR2, Markina OV2, Chemisova OS2, Pisanov RV2, Kruglikov VD2, Shestialtynova IS2, Uskova NN2
-
Affiliations:
- Rostov-on-Don Research Institute for PlagueControl, Russia
- Issue: Vol 89, No 2 (2012)
- Pages: 17-23
- Section: Articles
- Submitted: 09.06.2023
- Published: 15.06.2012
- URL: https://microbiol.crie.ru/jour/article/view/13657
- ID: 13657
Cite item
Full Text
Abstract
and study of its immunochemical properties.
Materials and methods. Rugose variants of
strains V. eltor 18895 and V. eltor 18843 obtained
by us by selection in M9 medium were used in
the study. Exopolysaccharides (EPS) were isolated
by K. Kierek (2003), S.P. Zadnova (2004),
N.P. Elinova (1984) methods and analyzed for
carbohydrate, protein, nucleic acid content and
lipopolysaccharide impurity. EPS, LPS, R-LPS
structure was compared by high-pressure chromatography.
Neutral sugars and amino sugars
were identified by thin layer chromatography.
Polyclonal antibodies were produced against
EPS preparation isolated by N.P. Elinova (1984)
method. Specific activity of obtained mice sera
was tested by DIA method. Results. EPS isolated
by N.P. Elinova method (1984) was shown
not to contain extraneous impurities. V. eltor
EPS structure differs from LPS and R-LPS.
Monosaccharide composition of EPS from ctx+
V. eltor 18895 strain is presented by a wider
specter of carbohydrates including glucose,
mannose, rhamnose, galacturonic acid. Use in
DIA of specific sera produced against EPS from
toxigenic strain did not reveal general epitopes
with capsule polysaccharides of V. cholerae
O139, V. parahaemolyticus and V. vulnificus.
Conclusion. Use of EPS as an immunogen promoted
production of sera that are specific against
EPS and rugose variants of Vibrio cholerae eltor
that can be used for their detection or characterization.
About the authors
O A Tatarenko
L P Alekseeva
N R Telesmanich
O V Markina
O S Chemisova
R V Pisanov
V D Kruglikov
I S Shestialtynova
N N Uskova
O A Tatarenko
Rostov-on-Don Research Institute for PlagueControl, RussiaRostov-on-Don Research Institute for PlagueControl, Russia
L P Alekseeva
Rostov-on-Don Research Institute for PlagueControl, RussiaRostov-on-Don Research Institute for PlagueControl, Russia
N R Telesmanich
Rostov-on-Don Research Institute for PlagueControl, RussiaRostov-on-Don Research Institute for PlagueControl, Russia
O V Markina
Rostov-on-Don Research Institute for PlagueControl, RussiaRostov-on-Don Research Institute for PlagueControl, Russia
O S Chemisova
Rostov-on-Don Research Institute for PlagueControl, RussiaRostov-on-Don Research Institute for PlagueControl, Russia
R V Pisanov
Rostov-on-Don Research Institute for PlagueControl, RussiaRostov-on-Don Research Institute for PlagueControl, Russia
V D Kruglikov
Rostov-on-Don Research Institute for PlagueControl, RussiaRostov-on-Don Research Institute for PlagueControl, Russia
I S Shestialtynova
Rostov-on-Don Research Institute for PlagueControl, RussiaRostov-on-Don Research Institute for PlagueControl, Russia
N N Uskova
Rostov-on-Don Research Institute for PlagueControl, RussiaRostov-on-Don Research Institute for PlagueControl, Russia
References
- Елинов Н.П. Микробные экзополисахариды. Киев, Наукова думка, 1984.
- Заднова С. П., Исаев Н. Д., Коннов Н. П. и др. Биохимический анализ двух фенотипически различных клонов штамма Vibrio cholerae Дакка 35 О1. Пробл. особо опасных инф. Саратов, 2004, 1 (87): 42-45.
- Захарова И.Я., Косенко Л.В. Методы изучения микробных полисахаридов. Киев, Наукова Думка, 1982.
- Остерман Л.А. Методы исследования белков и нуклеиновых кислот. Электрофорез и ультрацентрифугирование (практическое пособие). М. Наука, 1981.
- Спирин А.С. Определение нуклеиновых кислот. Биохимия. 1958, 23 (5): 656-662.
- Телесманич Н.Р. Анализ гемолизинов холерных вибрионов с помощью монослойных перевиваемых клеточных культур и моноклональных антител. Афтореф. дис. канд. биол. наук. Ростов-на-Дону, 1996.
- Galanos C., Luderitz O., Westphal O. A new method for the extraction of R lipopolysaccharides. Eur. J. Biochem. 1969, 9: 245-249.
- Kierek K., Watnick P.I. Environmental determinants of Vibrio cholerae biofilm development. Appl. Environ. Microbiol. 2003, 69 (9): 5079-5088.
- Lauriano C.M., Klose K.E., Kapfhammer D. Characterization of Vibrio cholerae O1 El Tor galU and gale mutants: influence on lipopolysaccharide structure, colonization and biofilm formation. Infect. Immun. 2001. 69 (1): 435-445.
- Nesper J., Kapfhammer D., Klose K. E. Characterization of Vibrio cholerae O1 antigen as bacteriophage K 139 receptor and identification of IS1004 insertions aborting O1 antigen biosynthesis. J. Bacteriol. 2000, 182: 5097-5104.
- Wai S.N., Mizunoe Y., Takade A. et al. V. cholerae O1 strain TSI-4 produces the exopolysaccharide materials determine colony morphology, stress resistance, and biofilm formation. Appl. Environ. Microbiol. 1998, 64 (10): 3648-3655.
- Westphal O., Jann K. Bacterial lipopolysaccharides. Extraction with phenol-water and further applications of the procedure. Carbohydr. Chem. 1965, 5: 83-91.