期刊文献+

CWPS竞争ELISA检测方法的建立 被引量:2

Development of competitative ELISA for quantitation of pneumococcal cell wall polysaccarides
原文传递
导出
摘要 目的:建立竞争酶联免疫吸附法(enzyme-linked immunosorbent assay,ELISA)测定C-多糖(Cell Wall Polysaccharides,cwes)浓度的方法。方法:以C-多糖为包被抗原,待测多糖为竞争抗原,与优化的抗C-多糖抗体反应,建立标准曲线,并验证该方法的准确性和精密度。结果:优化后的C-多糖包被浓度为10μg·mL^-1,抗C-多糖血清稀释度为1:160000。检测线性范围1250~20 ng·mL^-1,最低检测限为10ng·mL^-1,回归方程为B/B0=-0.2521 lg[CWPS]+0.9529,线性相关系数为R^2=0.99。批内和批间精密度分别为9.7%和6.9%,测定PS6A精糖中标准CWPS的回收率为100.3%。结论:本研究建立的竞争ELISA方法,准确性和精密度均较好,可以检测肺炎链球菌荚膜多糖中CWPS的浓度。 Objective: To develop competitive ELISA for determination of pneumococcal cell wall polysaccharides (CWPS) concentration. Methods: The standard curve was established by using CWPS as coating antigen, competing with polysaccharide in samples to react against antisera of pneumococcal ce The accuracy and precision of the method were validated accordingly. Results: The results 11 wall polysaccharides. showed that the optimal coating concentration of CWPS and serum dilution factor was 10 μg·mL^-1, and 1:160 000 respectively. The relation between CWPS concentrations and their reaction was linear in the range of 20~1 250 ng· mL^-1 CWPS concentration with the lowest limit of detection being 10 ng· mL^-1. The regression equation was B/B0-0.252 1 lg [CWPS] + 0.952 9 with RSQ = 0.99. The coefficient of variation of intra-assay and inter-assay was lower than 9.7% and 6.9%, respectively. The recovery rate of CWPS in pneumococcal polysaccharide serotype 6A was 100.3%. Conclusion: The competitive ELISA method developed showed high accuracy and precision. The method can be used for detection of CWPS in pneumococcal polysaccharide
出处 《中国新药杂志》 CAS CSCD 北大核心 2013年第21期2492-2495,共4页 Chinese Journal of New Drugs
基金 国际科技交流与合作专项(2010DFB33450)
关键词 CWPS 肺炎球菌 免疫学检测法 竞争ELISA cell wall polysaccharides pneumococcus immunoassay competitive enzyme-linked immu-nosorbent assay
  • 相关文献

参考文献18

  • 1KAYHTY H , ESKOLA J. New vaccines for the prevention ofpneumococcal infections [ J ]. Emerg Infect Dis, 1996 ,2 (4):289 -298.
  • 2林铃,朱为.肺炎球菌疫苗新策略[J].国际生物制品学杂志,2006,29(1):5-7. 被引量:13
  • 3SPANJAARD L, VANDER ENDE A.RUMKE H,e( al. van Al-phen. Epidemiology of meningitis and bacteraemia due to Strepto-coccus pneumoniae in The Netherlands[ J]. Acta Paediatr Suppl,2000,89(435) :22 -26.
  • 4GRAY BM,TURNER ME,DILLON HC JR. Epidemiologic stud-ies Streptococcus pneumoniae in infants. The effects of season andage on pneumococcal acquisition and carriage in the first 24months of life[ J]. Am J Epidemiol,1982,116(4) :692 - 703.
  • 5唐静,叶强.肺炎球菌疫苗的研究进展[J].国际呼吸杂志,2010,30(22):1392-1395. 被引量:6
  • 6LEE CJ, WANG TR. Pneumococcal infection and immunizationin children[ J]. Crit Rev Microbiol,1994 ,20(1) :1 -12.
  • 7朱继珩,蔡映云.肺炎球菌疫苗的临床应用[J].上海预防医学,2006,18(12):596-597. 被引量:1
  • 8BORGANO JM, MCLEAN AA,VELLA PP,ei al. Davidson.Vaccination and revaccination with polyvalent pneumococcal poly-saccharide vaccines in adults and infants [ J ]. Proc Soc Exp Biol,1978,157(1) :148 - 154.
  • 9SKOVSTED IC,KERRN MB, SONNE-HANSEN J,et al. Purifi-cation and structure characterization of the active component inthe pneumococcal 22 F polysaccharide capsule used for adsorptionin pneumococcal enzyme-linked immunosorbent assays [ J ]. Vac-cine ,2007 ,25 (35 ) :6490 -6500.
  • 10李雪莲,汪丽.七价肺炎球菌结合疫苗预防婴幼儿肺炎的临床分析[J].重庆医学,2011,40(15):1523-1524. 被引量:7

二级参考文献89

共引文献34

同被引文献20

  • 1陆权.儿童社区获得性肺炎管理指南(试行)(上)[J].中华儿科杂志,2007,45(2):83-90. 被引量:606
  • 2TALAGA P, BELLAHMY L, MOREAU M. Quantitative determi-nation of C-polysaccharide in Streptococcus pneumoniae capsularpolysaccharides by use of high-performanceanion-exchange chro-matography with pulsed amperometricdetection [ J ]. Vaccine,2001 ,19(20 -22) :2987 -2994.
  • 3FISHER W,BEHR T, HARTMANN K,et al. Teichoic acid andlipoteichoic acid of Streptococcus pneumonia possess identicalchain structures. A reinvestigation of teichoic acid ( C-polysac-charide) [ J ]. Eur J Biochem ,1993,11(3) :851 -857.
  • 4KULAKOWSKA M,BRISS0N JR, GRIFFITH DW,ei al. High-resolution NMR spectroscopic analysis of the C-polysaccharide ofStreptococcus pneumoniae [ J ]. Can J Chem,1993,71 (5 ) : 644 -648.
  • 5沈荣,任珍芸.肺炎链球菌C多糖的纯化方法:中国,201010564102. 8[P]. 2011 -06 - 15.
  • 6KARLSSON C’JANSSON PE,SKOV SORENSEN UB. The pneu-mococcal common antigen C-polysaccharideoccurs in differentforms. Monosubstituted or disubstituted with phosphocholine [ J ].Eur J Biochem (1999 ,265 (3) :1091 - 1097.
  • 7MOREAU M,RICHARDS JC,PERRY MB. Application of high-resolutionNMR spectroscopy to the elucidation of the structure ofthespecific capsular polysaccharide of Streptococcus pneumoniaetype 7F[J]. Carbohydr to,1988 ,182( 1) ;79 -99.
  • 8HELASSA N, VOLLMER W, BREUKINK E,et al. The mem-brane anchor of penicillin-binding protein PBP2a from Streptococ-cus pneumoniae influences peptidoglycanchain length [ J ]. FEBS7,2012,279(11) :2071 -2081.
  • 9NEESER JR, SCHWEIZER TF. A quantitative determination bycapillarygas-liquid hromatography of neutral and amino sugars ( asO-methyloxine acetates), and a study on hydrolytic conditionsforglycoproteins and polysaccharides in order to increase sugarre-coveries[ J ]. Anal Biochem,1984,142(1) :58 -67.
  • 10黄镇,陈玉秋,李薇,向左云,吴凯,陶佳明,范荣坤,何建东,方祖群.CDAP活化多糖制备5型肺炎链球菌荚膜多糖-破伤风类毒素结合疫苗[J].中国生物制品学杂志,2010,23(5):493-495. 被引量:11

引证文献2

二级引证文献1

相关作者

内容加载中请稍等...

相关机构

内容加载中请稍等...

相关主题

内容加载中请稍等...

浏览历史

内容加载中请稍等...
;
使用帮助 返回顶部