上转发光免疫层析法检测乙肝病毒外膜大蛋白在不同类型慢性肝病中的临床意义

发布时间：2018-05-31 08:06

本文选题：乙肝病毒外膜大蛋白 + 上转发光免疫层析法　；参考：《浙江大学》2015年硕士论文

【摘要】：目的：建立上转发光免疫层析技术(UPT-LF)检测乙肝病毒外膜大蛋白(HBV-LP)的方法,和酶联免疫吸附测定(ELISA)法进行比较,并分析此法用于乙肝病毒外膜大蛋白(HBV-LP)在慢性乙型肝炎(CHB)、乙肝后肝硬化(LC)、乙肝原发性肝癌(PHCC)患者血清中水平变化及血清检测的临床价值。方法：利用上转发光材料UCP作为示踪剂结合免疫层析技术组装乙肝病毒外膜大蛋白定量试纸条。利用基于上转发光法的免疫层析检测技术(UPT)检测260例慢性乙型肝炎、190例乙肝肝硬化及45例乙肝原发性肝癌患者血清中乙肝病毒外膜大蛋白(HBV-LP)的含量；化学发光法检测乙肝E抗原(HBeAg)含量；实时荧光定量PCR法检测乙肝病毒核酸(HBV DNA)。另外选取60例健康体检者作为对照组,同时用上转发光免疫层析法和ELISA法检测其血清HBV-LP的含量。分析乙肝病毒外膜大蛋白(HBV-LP)检测在不同疾病状态的诊断中的意义。结果： 1.成功建立基于上转发光免疫层析技术定量检测HBV-LP的试纸条；产品相关性能检测结果良好,最低检测限约为0.1U/ml, cut-off值约为1.0U/ml,批内与批间CV%均小于15%。 2.60例健康体检人员血清经UPT-LF法检测,IHBV-LP含量均小于1U/ml,特异性为100%,ELISA法检测结果显示59例血清标本HBV-LP阴性,另一例阳性,特异性为98.3%。 3.495例乙肝患者血清标本同时采用UPT-LF法与ELISA法检测HBV-LP,两种方法检测总符合率为91.7%,UPT法检测HBV-LP阳性率为58.5%,ELISA法检测HBV-LP阳性率为59.5%。 4.HBV-LP阳性结果浓度水平CHB组主要以40U/ml的高浓度为主,LC组和PHCC组主要集中在10U/ml的低浓度范围内。 5.HBeAg、HBV DNA和HBV-LP的阳性率均表现为CHB组LC组PHC组,HBeAg和HBV DNA在LC和PHCC患者中的阳性率均明显低于CHB患者,差异有统计学意义(P0.01)。 6.HBV-LP在LC和PHCC患者中的阳性率明显高于HBV DNA和HBeAg,差异有统计学意义(P0.05) 结论：上转发光免疫层析法检测血清HBV-LP简便、快捷、可靠,特异性优于ELISA法,并且HBV-LP的检测对于慢性肝病进展的诊断以及疗效监测有重要的价值。
[Abstract]:Objective : To establish a method for the detection of hepatitis B virus ( HBV - LP ) by up - to - light immunochromatography ( UPT - LF ) and compare it with enzyme - linked immunosorbent assay ( ELISA ) , and to analyze the clinical value of HBV - LP in the serum of patients with chronic hepatitis B , hepatitis B , liver cirrhosis ( LC ) , and primary liver cancer ( PHCC ) .

Methods : HBV - LP was detected in 260 patients with chronic hepatitis B , 190 cases of hepatitis B cirrhosis and 45 cases of primary hepatic carcinoma with hepatitis B .
The content of HBeAg was detected by chemiluminescence method .
HBV DNA was detected by real - time fluorescence quantitative PCR . In addition , 60 healthy subjects were selected as control group , and the contents of serum HBV - LP were detected by using up - to - light immunochromatography and ELISA . The significance of HBV - LP in the diagnosis of different disease states was analyzed .

Results :

1 . successfully establishing a test strip for quantitatively detecting HBV - LP based on an upconversion luminescence immune chromatography technique ;
The product - related performance test results are good , the lowest detection limit is about 0.1U / ml , the cut - off value is about 1.0U / ml , and the CV % between the batch and the batch is less than 15 % .

2.60 healthy persons were detected by UPT - LF method , the content of IHBV - LP was less than 1U / ml , the specificity was 100 % , the results of ELISA showed that HBV - LP was negative in 59 serum samples , and the other was positive , and the specificity was 98 . 3 % .

3.495 cases of HBV - LP were detected by UPT - LF and ELISA . The positive rate of HBV - LP was 58.5 % , and the positive rate of HBV - LP was 59.5 % .

4 . The concentration of HBV - LP positive result was mainly at 40U / ml high concentration , and the LC group and PHCC group were mainly concentrated in the low concentration range of 10 U / ml .

5 . The positive rates of HBeAg , HBV DNA and HBV - LP were significantly lower in the patients with LC , PHC , HBeAg and HBV DNA than those in the patients with LC and PHCC ( P0.01 ) .

6 . The positive rate of HBV - LP in patients with LC and PHCC was significantly higher than that in HBV DNA and HBeAg ( P0.05 ) .

Conclusion : The detection of HBV - LP is simple , rapid , reliable and better than ELISA in the detection of HBV - LP , and the detection of HBV - LP is of great value for the diagnosis of chronic liver disease progression and the monitoring of curative effect .
【学位授予单位】：浙江大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：R446.6

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