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通过酶联免疫吸附测定法检测动物和囊性纤维化患者血清中抗铜绿假单胞菌藻酸盐胞外多糖抗体。

Detection of antibodies to Pseudomonas aeruginosa alginate extracellular polysaccharide in animals and cystic fibrosis patients by enzyme-linked immunosorbent assay.

作者信息

Bryan L E, Kureishi A, Rabin H R

出版信息

J Clin Microbiol. 1983 Aug;18(2):276-82. doi: 10.1128/jcm.18.2.276-282.1983.

Abstract

An enzyme-linked immunosorbent assay was developed to detect antibodies to sodium alginate exopolysaccharide purified from three strains of Pseudomonas aeruginosa and commercial alginate from seaweed. Good attachment of alginate occurred with polystyrene microtiter plates at pH 7.0 with 0.04 M sodium phosphate buffer. With the enzyme-linked immunosorbent assay procedure, antibodies to alginate (not previously shown to be immunogenic) could be shown in humans and after immunization of mice and rabbits. Antibody to one of the alginates cross-reacted with two other P. aeruginosa alginates and commercial seaweed alginate. In animal immunization antibody titers were maximal after a single intravenous injection with an optimal dose of P. aeruginosa 3064 alginate. Healthy controls not known to have a previous P. aeruginosa infection had low, but detectable, antibody titers to 3064 and commercial alginate. Three cystic fibrosis patients not colonized with P. aeruginosa had similar antibody levels. Twenty-eight cystic fibrosis patients colonized with P. aeruginosa formed a clearly separate group with antibody titers higher than that of the control and noncolonized cystic fibrosis patients. Antibody titers to 3064 or commercial alginate did not increase during acute P. aeruginosa bronchopneumonitis in 16 cystic fibrosis patients or after repeated episodes in 4 patients.

摘要

开发了一种酶联免疫吸附测定法,用于检测从三株铜绿假单胞菌纯化的藻酸钠胞外多糖以及海藻商业藻酸盐的抗体。在pH 7.0的0.04M磷酸钠缓冲液中,藻酸盐能很好地附着在聚苯乙烯微量滴定板上。通过酶联免疫吸附测定法,可以在人类以及小鼠和兔子免疫后检测到针对藻酸盐的抗体(此前未显示具有免疫原性)。一种藻酸盐的抗体与另外两种铜绿假单胞菌藻酸盐和商业海藻藻酸盐发生交叉反应。在动物免疫中,单次静脉注射最佳剂量的铜绿假单胞菌3064藻酸盐后抗体滴度最高。已知既往无铜绿假单胞菌感染的健康对照对3064藻酸盐和商业藻酸盐的抗体滴度较低,但可检测到。三名未感染铜绿假单胞菌的囊性纤维化患者抗体水平相似。28名感染铜绿假单胞菌的囊性纤维化患者形成了一个明显不同的组,其抗体滴度高于对照组和未感染的囊性纤维化患者。在16名囊性纤维化患者的急性铜绿假单胞菌支气管肺炎期间或4名患者反复发病后,针对3064藻酸盐或商业藻酸盐的抗体滴度没有增加。

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