5种大肠杆菌O抗原定型血清的制备
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摘要
本研究以大肠杆菌参考菌株CVCC1369(O26)、CVCC3800(O153)、CVCC1489(O157)、CVCC3739(O159)、CVCC3807(O163)为菌种制备免疫抗原并免疫家兔,获得5种大肠杆菌O抗原定型原血清,而后制备多种吸收抗原以消除5种原血清的非特异性凝集素。通过微量凝集试验的方法,确定各血清的凝集价。最终确定了5种大肠杆菌O抗原定型血清的制备方法,为进一步改进大肠杆菌O抗原定型血清的制备工艺提供了思路和依据。
Antigens were prepared by using E.coli reference strains of CVCC1369(O26)、CVCC3800(O153)、CVCC1489(O157)、CVCC3739(O159) and CVCC3807(O163)respectively.And then 5 kinds of E. coli specific anti-O-antigen raw sera were obtained from rabbits immunized with the above antigens separately. Then cross-agglutinin was eliminated by non-specific absorption antigens. Cross-agglutination was detected by reacting sera with micro-agglutination test antigens respectively. Finally methods for the preparation of five kinds of E. coli specific anti-O-antigen serum were determined. Our study provides ideas and basis for further improving the preparation process of E. coli specific anti-O-antigen serum.
Antigens were prepared by using E.coli reference strains of CVCC1369(O26)、CVCC3800(O153)、CVCC1489(O157)、CVCC3739(O159) and CVCC3807(O163)respectively.And then 5 kinds of E. coli specific anti-O-antigen raw sera were obtained from rabbits immunized with the above antigens separately. Then cross-agglutinin was eliminated by non-specific absorption antigens. Cross-agglutination was detected by reacting sera with micro-agglutination test antigens respectively. Finally methods for the preparation of five kinds of E. coli specific anti-O-antigen serum were determined. Our study provides ideas and basis for further improving the preparation process of E. coli specific anti-O-antigen serum.
引文
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[4] 张媛,刘博,王秀丽,等.大肠杆菌菌体O 抗原单因子定型血清的制备[J].中国兽药杂志,2016,50(11):9- 16.
[5] 陆承平.兽医微生物学[M].北京:中国农业出版社,2007:97- 107.
[6] 章钢刚,赖卫华.食源性致病菌免疫学检测方法研究进展[J].食品安全质量检测,2015,6(9):3 414- 3 419.
[7] Fernando R P,Rezwanul W,Christina S F,et al.Serine protease autotransporters from Shigella flexneri and pathogenic Escherichia coli target a broad range of leukocyte glycoproteins[J].Proc Natl Acad Sci USA,2011,108(31):12 881- 12 886.
[8] 韩东雷.利用大肠杆菌合成糖蛋白并对糖链长短的调节进行初步探索[D].济南:山东大学,2015.
[9] Debroy C,Fratamico P M,Yan X,et al.Comparison of O-antigen gene clusters of all O-serogroups of Escherichia coli and proposal for adopting a new nomenclature for O-typing[J].PLoS one,2016,11(1):e0147434.
[10] GB/T 4789.6-2003.食品卫生微生物学检验致泻大肠埃希氏菌检验[S].
[11] 董伟.大肠杆菌血清抗体与硫酸庆大霉素偶联构建抗体靶向药物的研究[D].长沙:湖南农业大学,2007.
[2] 李晨,张双翔,周碧君,等.腹泻仔猪肠道致病性大肠埃希氏菌和沙门氏菌的分离鉴定与耐药性研究[J].中国畜牧兽医,2017,44(12):3 625- 3 633.
[3] 冯敏燕,林树乾,房玉波,等.山东省鸡源致病性大肠杆菌O-抗原混合血清型分析[J].中国兽医学报,2017,37(9):1 676- 1 679.
[4] 张媛,刘博,王秀丽,等.大肠杆菌菌体O 抗原单因子定型血清的制备[J].中国兽药杂志,2016,50(11):9- 16.
[5] 陆承平.兽医微生物学[M].北京:中国农业出版社,2007:97- 107.
[6] 章钢刚,赖卫华.食源性致病菌免疫学检测方法研究进展[J].食品安全质量检测,2015,6(9):3 414- 3 419.
[7] Fernando R P,Rezwanul W,Christina S F,et al.Serine protease autotransporters from Shigella flexneri and pathogenic Escherichia coli target a broad range of leukocyte glycoproteins[J].Proc Natl Acad Sci USA,2011,108(31):12 881- 12 886.
[8] 韩东雷.利用大肠杆菌合成糖蛋白并对糖链长短的调节进行初步探索[D].济南:山东大学,2015.
[9] Debroy C,Fratamico P M,Yan X,et al.Comparison of O-antigen gene clusters of all O-serogroups of Escherichia coli and proposal for adopting a new nomenclature for O-typing[J].PLoS one,2016,11(1):e0147434.
[10] GB/T 4789.6-2003.食品卫生微生物学检验致泻大肠埃希氏菌检验[S].
[11] 董伟.大肠杆菌血清抗体与硫酸庆大霉素偶联构建抗体靶向药物的研究[D].长沙:湖南农业大学,2007.
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