念珠菌优势抗体检测方法的建立及临床诊断价值评估  

Establishment of a novel serological test for dominant antibodies against Candida and assessment of its clinical diagnostic value

作　　者：马春芳[1] 陆静芬[1] 孔倩倩[1] 韩丹丹[1] 廖红[1] 李芳秋[1] 

机构地区：[1]南京军区南京总医院解放军临床检验医学研究所,南京医学硕士研究生210002

出　　处：《医学研究生学报》2013年第11期1138-1141,共4页Journal of Medical Postgraduates

基　　金：江苏省科技支撑计划-社会发展项目(BE2009673);南京军区医学科技创新重点课题(10Z027)

摘　　要：目的侵袭性念珠菌病(invasive candidiasis,IC)是免疫功能损伤或低下患者常见的感染和死亡原因,近年来检测IC患者血清中特异性抗体的血清学诊断方法成为研究热点。本文旨在建立检测念珠菌烯醇化酶(Enolase,Eno)、果糖二磷酸醛羧酶(fructose-bisphosphate aldolase,Fba1)特异性抗体的ELISA方法,评估其在IC早期诊断中的应用价值。方法利用基因重组技术获得的Eno、Fba1重组蛋白作为包被抗原,建立测定人血清相应抗体的ELISA,并对方法的敏感性、特异性进行考查。结果建立了检测IC患者血清抗Eno、抗Fba1抗体的ELISA,cut-off值分别为吸光度(A)值0.370和0.610。ELISA结果显示,IC患者血清中的抗Eno、抗Fba1抗体水平明显高于健康对照和非IC患者对照(P<0.001)。抗Eno和抗Fba1对诊断IC的敏感性和特异性分别为72.3%(73/101)和98%(196/200),87.1%(88/101)和96%(192/200),联合检测可提高敏感性至91.0%。且与细菌感染及其他真菌感染患者(如曲霉病)无交叉反应。IC患者中40.6%(41/101)可以在血培养阳性前检测到抗Eno、49.5%(50/101)可以在血培养阳性前检测到抗Fba1。结论建立了检测抗Eno、抗Fba1抗体的ELISA法,检测念珠菌菌丝相分泌蛋白IgG抗体特异性和灵敏度良好,在念珠菌感染诊断中具有潜在临床应用价值。Objective Invasive candidiasis （IC） is the major cause of morbidity and mortality in critically ill and immuno- compromised patients. Recent researches focus on the detection of specific antibodies for the serodiagnosis of IC. This study aimed to establish an ELISA-based method to detect IgG antibodies against enolase （Eno） and fructose-bisphosphate aldolase （Fbal） , and as- sess its value in the early diagnosis of IC. Methods Using recombinant Eno and Fbal as coated antigens, we established an ELISA- based method to detect the levels of anti-Eno and anti-Fbal antibodies in the sera of IC patients. Then we determined the sensitivity and specificity of this assay. Results The cut-off values for the serum anti-Eno and anti-Fbal antibodies were set by receiver operat- ing characteristic curves （ROC） with A = 0. 370 for anti-Eno and A = 0. 610 for anti-Fbal in the new ELISA. The median absorbance values of the anti-Eno and anti-Fbal antibodies were significantly higher in the serum of the IC patients than in that of the healthy indi- viduals （P 〈0.001 ）. The sensitivity and specificity of detection were 72.3% and 98% for anti-Eno and 87.1% and 96% for anti- Fbal antibodies, respectively. Combined detection of anti-Eno and anti-Fbal raised the sensitivity to 91%. Little crossreaction was found in patients with bacterial or other fungal infections. Positive antibody tests were shown in 40.6% （anti-Eno） and 49.5% （ anti- Fbal ） of the patients before positive blood cultures. Conclusion An ELISA-based method for detecting antibodies against Eno and Fbal was successfully developed, which has a potential clinical application value in the diagnosis of IC.

关 键 词：念珠菌 烯醇化酶 果糖二磷酸醛羧酶 抗体 ELISA 

分 类 号：R392.11[医药卫生—免疫学]