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鼠疫耶尔森菌rV抗原单抗系统及其ELISA检测方法的建立 被引量:5

Development of monoclonal antibodies and ELISA in detection of Yersinia pestis rV antigen
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摘要 目的建立ELISA双抗体夹心法,测定重组毒力因子rV抗原含量。方法采用杂交瘤技术,制备鼠疫菌rV抗原的鼠单克隆抗体,对抗原表位和单抗特异性进行分析及鉴定,建立ELISA双抗体夹心法,并验证方法的专属性、准确性、精密度和线性范围。结果成功组建了鼠疫菌rV抗原诊断试剂,灵敏度最低检测值为10 ng/mL。结论该方法可用于免疫学检测鼠疫组分疫苗原液rV抗原含量及制备过程中抗原活性,是鼠疫组分疫苗制备中一种重要的质量控制手段,也为进一步开发鼠疫诊断试剂盒及其他相关研究奠定了基础。 Objective To establish a sandwich ELISA in detection of the concentration of rV antigen of Yersinia pestis. Methods The McAbs against rV antigen were prepared by the hybridoma technology and Sandwich ELISA was developed on the basis of the epitope analysis and McAb specificity, in which specificity, accuracy, precision and linear range were v^idated. Results The diagnostic kit for Yersinia pestis rV antigen was successfully set up. The minimum detection con- centration of rV antigen is 10 ng/mL. Conclusion The kit is used in detection of the concentration of rV antigen in stock solution of plague component vaccine and in monitoring of antigeneicity in the preparing process. It is a important method for quality control in preparation of plague component vaccine, this method also provides a foundation for a further develo- ping the plague diagnostic reagents and other related investigations.
出处 《微生物学免疫学进展》 2012年第3期13-18,共6页 Progress In Microbiology and Immunology
基金 国家"863"课题(2006AA02Z461)
关键词 鼠疫耶尔森菌 rV抗原 单克隆抗体 表位 双抗体夹心ELISA Yersinia pestis rV antigen Monoclonal antibody (McAb) Epitope Sandwich ELISA
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  • 1Williamson ED, Eley SM, Griffin KF, et al. A new improved sub- unit vaccine for plague: the basis of protection[ J]. FEMS Immunol Med Microbiol, 1995, 12( 3/4 ): 223-230. 被引量:1
  • 2Leary SEC, Williamson ED, Griffin KF, et al. Active immuniza- tion with recombinant V antigen from Yersinia pestis protects mice against plague[ J]. Infect Immun, 1995, 63 ( 8 ) : 2854 -2858. 被引量:1
  • 3Andrews GP, Heath DG, Anderson GW, et al. Fraction 1 capsular antigen(F1 ) purification form Yersinia pestis CO92 and from an Escherichia coli recombinant strain and efficacy against lethal plague challenge [ J ]. Infect Immun, 1996,64 ( 6 ) :2180 - 2187. 被引量:1
  • 4Rocke TE, Smits S, Marinari P, et al. Vaccination with F1-V fusion protein protects black-footed ferrets (Mustela nigripes ) against plague upon oral challenge with Yersinia pestis [J]. J Wildl Dis, 2008, 44(1) :1 -7. 被引量:1
  • 5Koornhof H J, Smego JRA, Nicol M, et al. The Pathogenesis of Yersinia infection[J].Eur J Clin Microbial Infect Dis, 1999,18 (2) :87 - 112. 被引量:1
  • 6徐志凯.实用单克隆抗体技术[M].西安:陕西科学技术出版社,1991. 被引量:16
  • 7卢圣栋主编,李尹雄等编写..现代分子生物学实验技术[M].北京:高等教育出版社,1993:662.
  • 8焦奎,张书圣主编..酶联免疫分析技术及应用[M].北京:化学工业出版社,2004:281.

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  • 1徐秉臣,杨林,雷刚,热娜,阿扎提,王信惠.ELISA双McAb夹心法检测鼠疫F1抗原[J].地方病通报,1990,5(2):35-39. 被引量:8
  • 2Rahalison L, Vololonirina E, Ratsitorahina M, eta!. Diagnosis of bubonic plague by PCR in Madagascar under field conditions [J]. J Clin Microbial, 2000, 38 (1): 260-263. 被引量:1
  • 3Bianucci R, Rahalison L, Ferroglio E, et a/. A rapid diagnostic test for plague detects Yersinia pestis F1 antigen in ancient human remains [J]. Comptes Rendus Biol, 2007, 330 (10): 747-754. 被引量:1
  • 4Bianucci R, Rahalison L, Massa ER, et al. Technical note:a rapid diagnostic test detects plague in ancient human remains: an example of the interaction between archeological and biolo- gical approaches (southeastern France, 16th-18th centuries) [J]. Am J Phys Anthropol, 2008, 136 (3): 361-367. 被引量:1
  • 5Chanteau S, Rahalison L, Ratsitorahina M, et ol. Early diagnosis of bubonic plague using F1 antigen capture ELISA assay and rapid immunogold dipstick [J]. Int J Med Microbiol, 2000, 290 (3): 279-283. 被引量:1
  • 6Ratsitorahina M, Chanteau S, Rahalison L, et 01. Epidemio- logical and diagnostic aspects of the outbreak of pneumonic plague in Madagascar [J]. Lancet, 2000, 355(9198): 111-113. 被引量:1
  • 7Splettstoesser WD, Rahalison L, Grunow R, et 01. Evaluation of a standardized F1 capsular antigen capture ELISA test kit for the rapid diagnosis of plague [J]. FEMS Immunol Med Micr- obiol, 2004, 41 (2): 149-155. 被引量:1
  • 8蔡亦红,姚余有.3种食源性致病菌的多重PCR快速检测方法的建立[J].中国卫生检验杂志,2007,17(11):1959-1962. 被引量:16
  • 9Kim JS,Lee GG,Park JS,et al.A novel multiplex PCR assay for rapid and simultaneous detection of five pathogenic bacteria:Escherichia coli O157:H7,Salmonella,Staphylococcus aureus,Listeria monocytogenes,and Vibrio parahaemolyticus.J Food Prot,2007,70:1656-1662. 被引量:1
  • 10Notomi T,Okayama H,Masubuchi H,et al.Loop-mediated isothermal amplification of DNA.Nucleic Acids Res,2000,28:E63. 被引量:1

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