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实验性脑膜炎脑脊液中Ⅲ型肺炎链球菌荚膜多糖的酶免疫测定

Enzyme immunoassay of the capsular polysaccharide of Streptococcus pneumoniae type III in cerebrospinal fluid in experimental meningitis.

作者信息

Nolan C M, Ulmer W C

出版信息

J Med Microbiol. 1980 Nov;13(4):551-60. doi: 10.1099/00222615-13-4-551.

Abstract

An enzyme immunoassay (EIA) for the capsular polysaccharide of Streptococcus pneumoniae type III was developed and applied to the measurement of this antigen in cerebrospinal fluid (CSF) in an experimental model of pneumococcal meningitis. EIA was performed by a single-antibody sandwich technique in which the globulin fraction of pneumococcal type-specific antiserum (antiserum-globulin) was used to coat the solid phase before antigen attachment and to conjugate with the labelling enzyme, horseradish peroxidase. Under optimum assay conditions EIA detected purified pneumococcal type-III capsular polysaccharide in concentrations as low as 0.15 ng/ml in aqueous buffer. Assayed by EIA, the mean concentration of type-III capsular polysaccharide in CSF of rabbits with pneumococcal meningitis increased exponentially from 24 h to 96nh of infection (range 13.9 ng/ml--62 500 ng/ml). Effective antimicrobial therapy of rabbits with meningitis was associated with a rapid decrease in the CSF concentration of the capsular antigen, although it was still detected in concentration in the range 1--10 ng/ml in 100% of animals treated for 4 days. Thus EIA provides a quantitative and extremely sensitive method of measuring type-III pneumococcal capsular polysaccharide in CSF.

摘要

开发了一种用于检测Ⅲ型肺炎链球菌荚膜多糖的酶免疫测定法(EIA),并将其应用于肺炎球菌性脑膜炎实验模型中脑脊液(CSF)中该抗原的测量。EIA采用单抗体夹心技术进行,其中肺炎球菌型特异性抗血清(抗血清球蛋白)的球蛋白部分在抗原附着前用于包被固相,并与标记酶辣根过氧化物酶结合。在最佳测定条件下,EIA在水性缓冲液中能检测到低至0.15 ng/ml浓度的纯化Ⅲ型肺炎球菌荚膜多糖。通过EIA测定,患有肺炎球菌性脑膜炎的兔子脑脊液中Ⅲ型荚膜多糖的平均浓度在感染后24小时至96小时呈指数增加(范围为13.9 ng/ml - 62500 ng/ml)。对患有脑膜炎的兔子进行有效的抗菌治疗与脑脊液中荚膜抗原浓度的快速下降有关,尽管在接受4天治疗的所有动物中,仍能检测到浓度在1 - 10 ng/ml范围内的抗原。因此,EIA提供了一种定量且极其灵敏的方法来测量脑脊液中Ⅲ型肺炎球菌荚膜多糖。

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