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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(x2=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(x2=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(x2=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(x2=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 detect the anti-Pj rMsg IgG of 176 HIV-1 infected patients by ELISA using the recombinant major surface glycoprotein (rMsg) of Pneumocystis sp. IgM antibody detection rate. Methods The 350 bp fragment of the 5 ’end of the major surface glycoprotein gene was amplified by using Pj DNA obtained from clinical practice. The PCR product was cloned into the prokaryotic expression vector pGEX-3X to obtain GST-tagged rMsg was used to establish an ELISA method to detect the serum samples from 176 cases of HIV-1 infected patients in Beijing You’an Hospital. The detection rates of anti-Pj rMsg IgG and IgM antibodies were calculated. Results The positive rates of anti-Pj rMsg IgG and IgM antibodies in 176 HIV-1 infected patients were 24.4% (43/176) and 12.5% (22/176), respectively. Among them, IgG positive IgM, IgG positive IgM negative, IgG The positive rates of negative IgM and IgG negative IgM were 7.4% (13/176), 17.0% (30/176), 5.1% (9/176) and 70.5% (124/176), respectively. The positive rates of IgG antibodies in the age group of 20 to 29 years old, 30 to 39 years old and over 40 years old were 19.2% (10/52), 32.1% (25/78) and 17.4% respectively, (8/46). The positive rates of IgM were 11.5% (6/52), 12.8% (10/78) and 13.0% (6/52) 46). There was no significant difference between the two groups (x2 = 0.064, P = 0.969). In the CD + 4T lymphocyte count group, the positive rates of IgG antibody in ≥350 / μL and <350 / μL groups were 24.4% (30/123) and 24.5% (13/53) respectively, with no difference between the two groups The positive rates of IgM antibody were 12.2% (15/123) and 13.2% (7/53) respectively, there was no significant difference between the two groups (χ2 = 0.035, P = 0.984) 0.852). 22 cases of HIV-1 infected with anti-Pj rMg IgM antibody were clinically free of the symptoms and signs of pneumonia. Conclusions The detection rates of anti-Pj rMsg IgG and IgM antibodies in HIV-1 infected individuals are 24.4% and 12.5%, respectively. The seropositive evidence of Pj infection in this population is 29.5% positive for anti-Pj rMsg IgG or IgM. Anti-Pj rMsg IgM antibody positive can not be used as an indicator of Pj infection.