HBsAg确证实验与CHB患者血清miRNA-122等指标的分析应用

发布时间：2018-10-16 09:46

【摘要】：[摘要]目的探讨初诊HBsAg检测阳性患者标本进行抗体中和确证实验的研究；观察在CHB四个不同发病时期中,人外周血清中肝脏特异性miRNA-122表达量的变化和意义；探讨血清HBV-DNA低复制的CHB患者相关指标的变化及意义。方法收集CLIA法定量检测HBsAg阳性标本1974例,应用GICA、ELISA等方法复检HBsAg并进行统计学分析；此外,对CLIA法HBsAg阳性而ELISA法阴性的标本进行了CLIA法中和确证实验。收集80例CHB患者标本,采用FQ-PCR法和CLIA等方法对miRNA-122、HBV-DNA、HbeAg、HbsAg和ALT进行测定,对结果进行比较和线性相关分析。收集HBV-DNA低复制的CHB患者标本229例,进行mIL-2R、IL-10、miRNA-122等指标进行比较分析。结果对CLIA法检测的HBsAg阳性的标本使用GICA和ELISA两种方法进行复检后发现,GICA和ELISA法检坝HBsAg均有漏检现象的发生,其漏检率分别占HBsAg阳性标本的9.1%(180/1974)和25.4%(501/1974),而这些漏检的标本主要集中在CMIA法HBsAg检测值≤10.0IU/ml的样本中,而该浓度范围标本占HBsAg阳性标本的比例高达26.90%(531/1974)。本文将180例CMIA法HBsAg阳性标本而ELISA法漏检的标本经CMIA法中和确证实验阳性率为96.67%(174/180)。其中有6例无法确认,通过HBV-DNA检测与两周后随访确认3例为乙肝病毒感染病例,另3例排除。miRNA-122在CHB患者四个时期表达量均明显升高,尤其在免疫耐受期和免疫清除期升高显著(P0.01)；miRNA-122与HbeAg和HBV-DNA相关性不强(r=0.263,r=0.171),与HbsAg和ALT相关性较强(r=0.597,r=0.767)。低复制CHB患者中e抗原阳性患者占38.00%,e抗原阴性患者占51.53%；229例患者ALT升高者占总数的66.81%,患者血浆miRNA-122的表达量的变化与ALT浓度呈正相关(r=0.841)。e抗原阳性与e抗原阴性组IL-10、mIL-2R浓度与对照组比较均有统计学差异(P0.05)。结论ELISA等定性方法在低浓度HBsAg标本的检测上与CLIA等定量方法相比存在明显不足。实验室对首诊乙肝HBsAg检测的标本应首选检测灵敏度高的CLIA法进行检测,对低浓度HBsAg阳性患者应通过采集双份血清、中和试验、HBV-DNA检测以及随访建立档案等予以确认,避免漏诊误诊。miRNA-122和乙肝病毒复制可能没有关系,但对于乙肝的辅助诊断、评估及健康管理等具有重要的意义.HBV-DNA低复制CHB患者mIL-2R、 IL-10、miRNA-122等指标的变化证明此类患者部分病例仍存在肝细胞实质性的损害,临床应高度重视,尤其要与非活动性乙型肝炎病原携带状态相鉴别,不可一概而论。
[Abstract]:[abstract] objective to investigate the effect of antibody neutralization assay on HBsAg positive patients, and to observe the changes and significance of liver specific miRNA-122 expression in human peripheral serum during the four different stages of CHB. Objective: to investigate the changes and significance of serum HBV-DNA in CHB patients with low replication. Methods 1974 HBsAg positive specimens were detected by CLIA method, and HBsAg was re-examined by GICA,ELISA and analyzed statistically. In addition, CLIA method was used to confirm the positive samples of CLIA HBsAg and negative ELISA. The samples of 80 patients with CHB were collected. The miRNA-122,HBV-DNA,HbeAg,HbsAg and ALT were measured by FQ-PCR and CLIA. The results were compared and analyzed by linear correlation. Samples of CHB patients with low replication of HBV-DNA were collected and compared with mIL-2R,IL-10,miRNA-122. Results the GICA and ELISA methods were used to re-examine the HBsAg positive specimens detected by CLIA method. The results showed that the HBsAg of the dam detected by GICA and ELISA methods were both missed. The rate of missed detection was 9.1% (180 / 1974) and 25.4% (501 / 1974) of HBsAg positive specimens, respectively. These missed samples were mainly concentrated in samples with HBsAg detection value 鈮，

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