FRACTIONATION OF PNEUMOCYSTIS-CARINII ANTIGENS USED IN AN ENZYME-LINKED IMMUNOSORBENT-ASSAY FOR ANTIBODIES AND IN THE PRODUCTION OF ANTISERUM FOR DETECTING PNEUMOCYSTIS-CARINII ANTIGENEMIA
FRACTIONATION OF PNEUMOCYSTIS-CARINII ANTIGENS USED IN AN ENZYME-LINKED IMMUNOSORBENT-ASSAY FOR ANTIBODIES AND IN THE PRODUCTION OF ANTISERUM FOR DETECTING PNEUMOCYSTIS-CARINII ANTIGENEMIA
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DOI：
10.1128/jcm.15.6.1029-1035.1982
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发表时间：
1982-01-01
影响因子：
9.4
通讯作者：
NORMAN, LG
中科院分区：
文献类型：
--
作者：
MADDISON, SE;HAYES, GV;NORMAN, LG
用差速和梯度离心法从严重感染的大鼠肺组织中获得了卡氏肺孢子虫的富囊悬液。在制备富含包囊物质的水溶提取物后，用8M尿素提取不溶残留物。少量感染的人肺组织和未感染的大鼠和人的肺被类似地处理。十二烷基硫酸钠-聚丙烯酰胺凝胶电泳显示，人和大鼠受感染的肺提取液含有大量蛋白质(>；200,000道尔顿)。这一成分在未感染肺的提取物中不存在。从感染大鼠肺的富含包囊的悬浮液中也制备了盐酸可溶提取物。在酶联免疫吸附试验中，尿素提取的抗原反应最强。用对流免疫电泳法检测患者血清中循环抗原。
Cyst-rich suspensions of P. carinii were obtained by differential and gradient centrifugation from heavily infected rat lungs. After preparation of an aqueous-soluble extract of the cyst-rich material, the insoluble residue was extracted with 8 M urea. Small amounts of infected human lung tissue and uninfected rat and human lung were processed similarly. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis showed that human and rat infected lung extracts contained a large protein (> 200,000 daltons). This component was not present in extracts of uninfected lung. An HCl-soluble extract also was prepared from the cyst-rich suspension from infected rat lung. The urea-extracted antigen was most reactive in an enzyme-linked immunosorbent assay. Rabbit antiserum against the HCl-soluble antigen detected circulating antigen in patients'' sera in a counterimmunoelectrophoresis assay.