Research on PCR determining of type b Haemophilus influenzae strains and immunogenicity of polysaccharide conjugates
10.3760/cma.j.issn.0254-5101.2012.04.015
- VernacularTitle:几株b型流感嗜血杆菌多糖结合物的免疫原性检定
- Author:
Yulan YUAN
;
Jingrong GUO
;
Jiwei ZHOU
;
Hua GAO
- Publication Type:Journal Article
- Keywords:
Haemophilus influenzae;
Sentype-specific gene;
Capsular type gene;
Capsular polysaccharide;
Immunogenicity
- From:
Chinese Journal of Microbiology and Immunology
2012;32(4):346-350
- CountryChina
- Language:Chinese
-
Abstract:
Objective To determine Haemophilus influenzae type b strains in molecular level using PCR,and to study the immunogenicity of capsular polysaccharide conjugates in mice.Methods Extracting genomes using bacterial DNA extract kit from Hoemophilus influenzae type b strains,and PCR for determining the strains through serotyping-specific and capsular genotyping primers respectively.Various capsular polysaccharides conjugated TT respectively,and the conjugates were administered subcutaneously to mice through dilution.After vaccination with two doses,blood samples were collected for the detection of antibody levels to polyribosylribitol phosphate ( PRP),the capsular polysaccharide of Hib.Results All five Haemophilus influenzae type b strains contain type-specific(482 bp) and capsular type (343 bp)DNA fragment through PCR detecting.The DNA fragments were sequenced.BLAST show that these sequences are 100％ homology comparing the above strains respectively,and are 99％ and 100％ homology comparing the GenBank X78559.1 and M19995.1 respectively.The immunogenicity of mice from various capsular polysaccharide conjugates (PRP-TT) was not significantly different by ELISA detecting.Conclusion Through PCR,Haemophilus influenzae type b strain can be determined in molecular level.The immunogenicity of mice from purified capsular polysaccharide conjugates was not different.The study provides a detection means for the features and heredity stability of Haemophilus influenzae type b strain and reference data for the immunogenicity of different polysaccharide conjugates in vaccine research and development and production.
