重组融合蛋白早期快速诊断侵袭性念珠菌病的实验研究  

Research on recombinant fusion protein in early fast diagnosis of invasive candidiasis

作　　者：陈晶[1] 

机构地区：[1]解放军白求恩国际和平医院检验实验科,河北石家庄050082

出　　处：《实用临床医药杂志》2015年第15期55-57,61,共4页Journal of Clinical Medicine in Practice

基　　金：河北省自然科学基金项目(C2010001882)

摘　　要：目的建立一种IC早期快速准确的诊断方法。方法 178例临床需要送检侵袭性念珠菌病常规检测和真菌培养的患者血清为研究对象,其中103例确诊为侵袭性念珠菌病者作为病例组,75例为在同样免疫妥协易感人群但无念珠菌感染微生物学证据的患者血清作为对照组。应用基因重组的方法,将念珠菌菌丝壁蛋白(HWP1),与位于细胞壁的糖酵解酶(烯醇化酶蛋白ENO1)进行基因克隆,随后将由HWP1与ENO1组成的重组蛋白在大肠杆菌中表达,并建立酶联免疫方法(ELISA),检测患者血清中的Ig G特异性抗体。通过统计学的分析,计算出该方法的灵敏度,特异性,阳性预测值,阴性预测值。筛选出灵敏度高,特异性强,阳性预测值和阴性预测值均高的融合蛋白抗原建立的ELISA方法。结果病例组中抗-ENO1、抗-HWP1以及二者联合检测的阳性检出率均显著高于对照组(P<0.01),差异有统计学意义。抗-HWP1与抗-ENO1两方法相比,抗-HWP1的敏感性(χ2=37.622,P<0.01)、阳预测值(χ2=74.427,P<0.01)及阴性预测值(χ2=2.057,P=0.152)均显著高于抗-ENO1;联合检测与抗-ENO1比较,敏感性(χ2=44.524,P<0.01)、特异性(χ2=9.765,P=0.002)及阳预测值(χ2=72.000,P<0.01)均显著高于抗-ENO1;联合检测与抗-HWP1相比,特异性(χ2=4.700,P=0.030)、阴性预测值(χ2=3.191,P=0.074)显著高于抗-HWP1。结论 ENO1和HWP1蛋白重组后联合诊断的效果更佳,可以作为IC早期快速准确的诊断方法。Objective To establish a early fast diagnostic method for invasive candidiasis. Methods A total of 178 patients＇ serums demanding laboratory detection of invasive candidiasis and fungus culture were selected as the research objects, among which 103 serum samples diag- nosed invasive candidiasis were included in the disease group and another 75 serum samples in the immunocompromised susceptible population but without monilial infection were as the control group. Candida albicans hyphal wall protein （HWP1） and glyeolytic enzyme （enolase protein ENO1） located in the cell wall were cloned by gene recombination, and the recombinant protein composed of HWP1 and ENO1 were expressed in Escherichia coli. Further more, enzyme linked immunosorbent assay （ELISA） was established to detect specificity antibodies of IgG in the serum of the patients. Statistical analysis was used to calculate the sensitivity, specificity, positive prediction value, and negative prediction value. Meanwhile, the fusion protein antigen with higher sensi- tivity, specificity, positive prediction value and negative prediction value was screened. Results The positive detection rates of anti -ENO1, anti -HWP1 and combined detection were significantly higher than that in control group, the difference was statistically significant （P 〈 0. 01 ）. Comparisomf Anti- HWP 1 and anti- ENO 1 showed that the sensitivityof anti- HWP 1 （ X2 = 3 7.6 2 2, P 〈 0. 0 1 ） , positive predictive value （ X2 = 7 4.4 2 7 , P 〈 0.01 ） and negative predictive value （ X2 = 2. 0 5 7, P = 0. 1 5 2 ） were significantly higher than that of anti - ENO 1. The sensitivity （X2 = 44. 524, P〈0.01） and specific of （X2= 9. 765, P = 0. 002） and positive predictive value （X2 = 72. 000, P〈 0.01 of joint detection were significantly higher than that anti-ENO1. The specificity （X2 = 4. 700, P = 0. 030） and negative predictive value （X2 = 3. 191, P = 0. 074） of joint detection was significantly higher than that of anti-HWPl. Conclusion Joi

关 键 词：侵袭性念珠菌病 HWP1 ENO1 重组蛋白 实验室诊断 

分 类 号：R519.3[医药卫生—内科学]