用芯片研究丙型肝炎病毒不同功能区抗体

Approach to Antibodies of Different Regions of HCV Genome and Its Clinical Significance with Protein Chip

目的 用蛋白质芯片研究 HCV不同功能区抗体及临床意义。方法 将抗原性强的 C、NS3、NS4、NS5片段用芯片点样仪定量点加在醛基化玻片上来制作蛋白芯片。用进口 HCV(EL ISA)筛选出 6 5例抗 - HCV阳性标本 ,阳性标本用芯片检测不同功能区抗体 ,并测定 HCV- RNA;同时用芯片检测 2 4例抗 - HCV阴性的正常献血员血清。结果 2 4例抗 - HCV阴性标本用芯片检测均阴性 ;6 5例抗 - HCV阳性标本经芯片均有 1～ 4种抗体阳性 ,其中抗 - C+抗 - NS3+抗 - NS4+抗 - NS5阳性率最高 ,占 33.8% ;抗 - C、抗 - NS3、抗 NS5、抗 NS4总的检出率分别为98.5 %、89.2 %、5 2 .3%和 5 0 .8% ;未发现单独含抗 - NS3、抗 - NS4的血清 ;抗 - C+抗 NS3+抗 - NS4+抗 - NS5阳性血清中 HCV- RNA检出率最高 ,占 77.3% ;HCV- RNA阳性血清中抗 - NS5阳性率最高 ,占 6 1.8% ,说明抗 - NS5与 HCV- RNA最有相关性 (P<0 .0 5 )。结论 蛋白质芯片显示较高的敏感性与特异性 ;抗 - C、抗 - NS3在诊断抗 -HCV都很重要 ,抗 - NS4及抗 - NS5有诊断的互补作用 ,而抗 - NS5在一定程度上能反映 HCV的复制情况。

OBJECTIVE To approach to antibodies of different regions of HCV genome and its clinical significance with protein chip. METHODS Strong antigenicity of C,NS3,NS4 and NS5 was quantitatively pointed on aldehydized glass slide with chip sampling apparatus to make chip.Sixty five cases of anti HCV positive specimens were screened out with generation of imported HCV ELISA kits. Antibodies to different regions of the positive specimens were detected with chip and HCV RNA was determined. Simultaneously,24 case donor sera with anti HCV negative were detected with chip.RESULTS Twenty four cases of specimens with anti HCV negative were all negative. Sixty five cases of specimens with anti HCV positive were one to four antibodies positive, from them, anti C+anti NS3+anti NS4+anti NS5 positive rate was the highest(33.8%).The total rates of anti C,anti NS3,anti NS5 and anti NS4 were 98.5%,89.2%,52.3% and 50.8%,respectively.Sole anti NS3 and anti NS4 were not detected. The sera with anti C+,anti NS3+and anti NS5+had the highest HCV RNA detected rate (77.3%).The sera with HCV RNA positive had the highest anti NS5 detected rate.There is a good correlation between anti NS5 and HCV RNA(P<0.05).CONCLUSIONS Protein chip showed relative sensitivity and specificity. Anti C and anti NS3 were very important to diagnose HCV, anti NS4 and anti NS5 had mutually complemented effect in diagnosis. Moreover, anti NS5 can reflect the replication situation of HCV to certain extent.