HIV-1感染者抗耶氏肺孢子菌主要表面糖蛋白抗体筛查  

Screening of anti-Pneumocystis jirovecii recombinant major surface glycoprotein antibodies in HIV-1 infected patients

作　　者：宋营改[1] 刘意[2] 任珊珊[1] 张彤[2] 范东瀛[3] 李威[2] 任翊[1] 

机构地区：[1]首都医科大学附属北京友谊医院、北京热带医学研究所,北京100050 [2]首都医科大学附属北京佑安医院,北京100069 [3]首都医科大学基础医学院病原生物学系,北京100069

出　　处：《中国艾滋病性病》2014年第10期711-715,共5页Chinese Journal of Aids & STD

基　　金：首都医科大学基础临床科研合作基金(14JL03)~~

摘　　要：目的利用原核表达的耶氏肺孢子菌（Pj）重组主要表面糖蛋白（rMsg）建立酶联免疫吸附试验（ELISA）方法,调查176例艾滋病病毒（HIV-1）感染者的抗Pj rMsg IgG和IgM抗体的检出率。方法以临床获得的Pj脱氧核糖核酸（DNA）为模板,扩增主要表面糖蛋白基因的5’端350bp片段,将聚合酶链反应（PCR）产物克隆到原核表达载体pGEX-3X中,获得可溶GST标签的rMsg,建立ELISA方法,检测北京佑安医院176例HIV-1感染者的血清标本,统计抗Pj rMsg IgG和IgM抗体的检出率。结果 176例HIV-1感染者中,抗Pj rMsg IgG和IgM抗体检出率分别为24.4%（43/176）和12.5%（22/176）,其中IgG阳性IgM阳性、IgG阳性IgM阴性、IgG阴性IgM阳性和IgG阴性IgM阴性的检出率,分别为7.4%（13/176）、17.0%（30/176）、5.1%（9/176）和70.5%（124/176）。抗体调查年龄分组中,（20~29）岁、（30~39）岁和40岁以上年龄组的IgG抗体阳性率分别为19.2%（10/52）、32.1%（25/78）和17.4%（8/46）,组间比较差异无统计学意义（x^2=4.450,P=0.108）;IgM阳性率分别为11.5%（6/52）、12.8%（10/78）和13.0%（6/46）,组间比较差异无统计学意义（x^2=0.064,P=0.969）。CD＋4T淋巴细胞计数分组中,≥350个/μL组和〈350个/μL组的IgG抗体阳性率分别为24.4%（30/123）和24.5%（13/53）,两组比较差异无统计学意义（x^2=0.000,P=0.984）;IgM抗体阳性率分别为12.2%（15/123）和13.2%（7/53）,两组比较差异无统计学意义（x^2=0.035,P=0.852）。抗Pj rMsg IgM抗体阳性的22例HIV-1感染者,临床没有肺炎的症状与体征。结论 HIV-1感染人群中抗Pj rMsg IgG和IgM抗体检出率分别为24.4%和12.5%,该人群感染Pj的血清学证据,抗Pj rMsg IgG或IgM阳性的检出率为29.5%。抗Pj rMsg IgM抗体阳性不能作为Pj现症感染的指标。Objective To establish an enzyme-linked immunosorbent assay（ELISA）with Pneumocystis jirovecii（Pj）recombinant major surface glycoprotein（rMsg）and investigate anti-Pj rMsg IgG and IgM antibodies in 176HIV-1infected patients from Beijing local hospital.Methods Using clinically acquired Pj DNA as the template,a fragment of 350 base pairs located at the 5＇end of Pj Msg gene was amplified and cloned into a prokaryotic expression vector of pGEX-3Xto obtain a GST-Msg fusion protein.An ELISA method was established to detect anti-Pj rMsg IgG and IgM antibodies in 176 serum samples from HIV-1infected patients in Beijing You＇an Hospital.Detection rates of anti-Pj rMsg IgG and IgM antibodies were analyzed.Results Positive rates of anti-Pj rMsg IgG and IgM antibodies in the 176 HIV infected patients were24.4%（43/176）and 12.5%（22/176）,respectively,among which rates of IgM positive IgG positive,IgG positive IgM negative,IgG negative IgM positive and IgG negative IgM negative were 7.4%（13/176）,17.0%（30/176）,5.1%（9/176）and 70.5%（124/176）,respectively.In respect of age groups,anti-Pj rMsg IgG antibody positive rates in 20-29 years age group,30-39 years age group and over 40 years age group were 19.2%（10/52）,32.1%（25/78）and17.4%（8/46）,respectively.There was no significant difference among these three groups（x^2=4.450,P=0.108）.Anti-Pj rMsg IgM antibody positive rates were 11.5%（6/52）,12.8%（10/78）and 13.0%（6/46）,respectively.There was no significant difference among the groups（x^2=0.064,P=0.969）.In respect of CD＋4T lymphocyte count groups,anti-Pj rMsg IgG antibody positive rates in group of CD＋4T lymphocytes≥350cells/μl and group of CD＋4T lymphocytes350cells/μL were 24.4%（30/123）and 24.5%（13/53）,respectively.There was no significant difference between the two groups（x^2=0.000,P=0.984）.IgM antibody positive rates were12.2%（15/123）and 13.2%（7/53）,respectively.There was no significant difference between the two groups（x^2=0.035,P=0.852

关 键 词：耶氏肺孢子菌 主要表面糖蛋白 重组抗原 抗体筛查 

分 类 号：R373.9[医药卫生—病原生物学]