白色念珠菌H3K4甲基转移酶N端肽段抗体的ELISA检测方法建立  

Evaluation of a Novel Enzyme-linked Immunosorbent Assay to Detect Specific Antibody to N-terminal Region of Histone 3 Lysine 4 Methyltransferase for Serodiagnosis of Invasive Candidiasis

作　　者：孔倩倩[1] 马春芳[1] 蔡迁[1] 廖红[1] 韩丹丹[1] 史利宁[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,南京210002

出　　处：《现代检验医学杂志》2013年第2期10-13,共4页Journal of Modern Laboratory Medicine

基　　金：基金项目：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（102027）.

摘　　要：目的建立检测人血清中抗白色念珠菌H3K4甲基转移酶N末端肽段（N—terminal region of histone 3 lysine 4 methyhransferase，Setl-208p）IgG类抗体的ELISA方法，评估其在侵袭性念珠菌病（invasive candidiasis，IC）患者中的早期诊断价值。方法收集IC患者（105例）、定植患者（37例）、细菌感染患者（25例）、其他真菌感染患者（10例）以及健康体检者（200例）血清，用Setl-208p作为包被抗原，血清1：500稀释，以羊抗人IgG-HRP为二抗，测定人血清中相应的抗Setl-208p抗体水平，确定cut-off值并考查方法的敏感度、特异度和准确度。结果以重组Setl-208p抗原建立的ELISA法精密度良好，批内变异系数（coefficient of variation，CV）为5．5％，批间CV为10．4％。重组抗原对血清中相应抗体的阻断率为91．8％，根据ROC曲线，选择吸光度（absorbance，A）值0．40作为cut off值，对IC的诊断敏感度为79．2％，特异度为90％，且与细菌感染及其他真菌感染病人（如曲霉）无非特异交叉反应。此外，IC患者中抗Setl-208p抗体阳性率（76．1％，80／105）显著高于念珠菌定植者抗Setl-208p抗体阳性率（32．4％，12／37）（χ^2=22．9，P〈0．01）。结论建立了检测抗白念珠菌抗Setl-208p抗体的ELISA法，在IC早期诊断中具有潜在应用价值。Objective To establish an ELISA method for detecting IgG antibody to N-terminal region of C. albicans histone 3 lysine 4 methyltransferase （Setl-208p） in human sera,and evaluate its value in diagnosis of invasive candidiasis （IC）. Meth- ods Sera from 105 cases of patients with IC,37 cases with Candida albicans colonization,25 cases with bacterial infections and 200 healthy subjects were collected. The ELISA method for detecting anti-Setl-208p antibody was established with recombinant Setl-208p as coating antigen. Patient sera were diluted in 1 ： 500,and peroxidase-conjugated goat anti-human immunoglobulin G （IgG） was added as the secondary antibody. The cut-off value,sensitivity and specificity of the assay was determined. Results The precision of ELISA was good with within-run CV 5.5 % and between-run CV 10.4% respectively. The blocking rate of sera with recombinant Setl-208p was 91.8%. According to ROC curve,the absorbance value of 0.40 was set as the cut off value. In this condition, the sensitivity, specificity of detection were 79.2 % and 90% for anti-Setl- 208p. Little crossreaction was found with patients with bacterial infection and other fungal infections such as Aspergillus. The positive rate of anti-Setl-208p antibody in patients with IC （76.1% ,80/105） was significantly higher than that of patients with Candida albicans colonization （32.4%, 12/37）（χ^2 =22.9,P〈0.01）. Conclusion An ELISA-hased method was developed for antibody against C. albicans setl-208p,which may had potential value for early diagnosis of IC.

关 键 词：念珠菌 H3K4甲基转移酶 抗体 ELISA 血清学诊断 

分 类 号：R379.4[医药卫生—病原生物学] R446.61[医药卫生—基础医学]