南方医科大学学报
南方醫科大學學報
남방의과대학학보
JOURNAL OF SOUTHERN MEDICAL UNIVERSITY
2014年
5期
646-650
,共5页
杨梅%王卓娅%郝卫%王艳芳%黄莉%蔡建飘%江凌晓%车小燕%钟晓祝%余楠
楊梅%王卓婭%郝衛%王豔芳%黃莉%蔡建飄%江凌曉%車小燕%鐘曉祝%餘楠
양매%왕탁아%학위%왕염방%황리%채건표%강릉효%차소연%종효축%여남
烟曲霉%双抗原夹心ELISA法%抗体检测%Afmp1p%Afmp1cr%Afmp2cr
煙麯黴%雙抗原夾心ELISA法%抗體檢測%Afmp1p%Afmp1cr%Afmp2cr
연곡매%쌍항원협심ELISA법%항체검측%Afmp1p%Afmp1cr%Afmp2cr
Aspergillus fumigatus%double- antigen sandwich enzyme- linked immunosorbent assay%antibody detection%Afmp1p%Afmp1cr%Afmp2cr
目的：建立检测烟曲霉特异性抗体的双抗原夹心ELISA方法。方法用毕赤酵母系统表达重组的烟曲霉特异性甘露聚糖蛋白片段Afmp1cr和Afmp2cr，经棋盘滴定法建立可检测烟曲霉特异性抗体的双抗原夹心ELISA方法。用Afmp1cr和Afmp2cr分别免疫兔血清评估方法灵敏度，健康人血清、其它微生物感染者血清评估特异性，并检测曲霉感染者血清。结果 Afmp1cr双抗原夹心ELISA法可捕获1∶800稀释兔多抗血清中的抗体，Afmp2cr双抗原夹心ELISA法可捕获1∶3200稀释兔多抗血清中的抗体，两者均与其他真菌及细菌无交叉反应。2例确诊曲霉感染和1例拟诊曲霉感染患者血清中能检测出抗体。结论初步建立了检测anti-Afmp1cr/Afmp2cr抗体的双抗原夹心ELISA方法，可辅助诊断临床烟曲霉感染。
目的：建立檢測煙麯黴特異性抗體的雙抗原夾心ELISA方法。方法用畢赤酵母繫統錶達重組的煙麯黴特異性甘露聚糖蛋白片段Afmp1cr和Afmp2cr，經棋盤滴定法建立可檢測煙麯黴特異性抗體的雙抗原夾心ELISA方法。用Afmp1cr和Afmp2cr分彆免疫兔血清評估方法靈敏度，健康人血清、其它微生物感染者血清評估特異性，併檢測麯黴感染者血清。結果 Afmp1cr雙抗原夾心ELISA法可捕穫1∶800稀釋兔多抗血清中的抗體，Afmp2cr雙抗原夾心ELISA法可捕穫1∶3200稀釋兔多抗血清中的抗體，兩者均與其他真菌及細菌無交扠反應。2例確診麯黴感染和1例擬診麯黴感染患者血清中能檢測齣抗體。結論初步建立瞭檢測anti-Afmp1cr/Afmp2cr抗體的雙抗原夾心ELISA方法，可輔助診斷臨床煙麯黴感染。
목적：건립검측연곡매특이성항체적쌍항원협심ELISA방법。방법용필적효모계통표체중조적연곡매특이성감로취당단백편단Afmp1cr화Afmp2cr，경기반적정법건립가검측연곡매특이성항체적쌍항원협심ELISA방법。용Afmp1cr화Afmp2cr분별면역토혈청평고방법령민도，건강인혈청、기타미생물감염자혈청평고특이성，병검측곡매감염자혈청。결과 Afmp1cr쌍항원협심ELISA법가포획1∶800희석토다항혈청중적항체，Afmp2cr쌍항원협심ELISA법가포획1∶3200희석토다항혈청중적항체，량자균여기타진균급세균무교차반응。2례학진곡매감염화1례의진곡매감염환자혈청중능검측출항체。결론초보건립료검측anti-Afmp1cr/Afmp2cr항체적쌍항원협심ELISA방법，가보조진단림상연곡매감염。
Objective To establish two double-antigen sandwich ELISA systems to detect anti-Afmp1cr and Afmp2cr antibodies of Aspergillus fumigatus. Methods Recombinant Afmp1cr and Afmp2cr proteins of A.fumigatus expressed in Pichia pastoris were obtained. Double-antigen sandwich ELISA systems for detecting specific anti-Afmp1cr and anti-Afmp2cr antibodies were developed after chessboard titrating to determine the appropriate concentrations of the recombinant proteins and HRP-labeled proteins. The sensitivity of the assay was evaluated using serum samples of rabbits immunized with Afmp1cr and Afmp2cr. The specificity of the assay was evaluated by detecting serum samples from healthy donors and patients with other pathogenic fungal and baterial infections. The performance of the two ELISA kits was furthered evaluated using serum samples from patients with suspected Aspergillus infection. Results The established ELISA kits were capable of detecting anti-Afmp1cr and anti-Afmp2cr antibodies in immunized rabbit serum at the maximum dilutions of 800 and 3200, respectively. No cross-reactivity was observed in detecting serum from patients with other pathogenic fungal or bactetial infections. Both of the two kits yielded positive results in sera from two established Aspergillus-infected cases and a suspected case. Conclusion Two antibody-capture ELISA kits were developed for the laboratory diagnosis of A.fumigatus infection and can be potentially useful in the clinical diagnosis of Aspergillosis infections.
