本文選題：乙肝病毒外膜大蛋白 + 上轉(zhuǎn)發(fā)光免疫層析法　； 參考：《浙江大學(xué)》2015年碩士論文

【摘要】：目的：建立上轉(zhuǎn)發(fā)光免疫層析技術(shù)(UPT-LF)檢測乙肝病毒外膜大蛋白(HBV-LP)的方法,和酶聯(lián)免疫吸附測定(ELISA)法進(jìn)行比較,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)、乙肝原發(fā)性肝癌(PHCC)患者血清中水平變化及血清檢測的臨床價(jià)值。 方法：利用上轉(zhuǎn)發(fā)光材料UCP作為示蹤劑結(jié)合免疫層析技術(shù)組裝乙肝病毒外膜大蛋白定量試紙條。利用基于上轉(zhuǎn)發(fā)光法的免疫層析檢測技術(shù)(UPT)檢測260例慢性乙型肝炎、190例乙肝肝硬化及45例乙肝原發(fā)性肝癌患者血清中乙肝病毒外膜大蛋白(HBV-LP)的含量；化學(xué)發(fā)光法檢測乙肝E抗原(HBeAg)含量；實(shí)時(shí)熒光定量PCR法檢測乙肝病毒核酸(HBV DNA)。另外選取60例健康體檢者作為對(duì)照組,同時(shí)用上轉(zhuǎn)發(fā)光免疫層析法和ELISA法檢測其血清HBV-LP的含量。分析乙肝病毒外膜大蛋白(HBV-LP)檢測在不同疾病狀態(tài)的診斷中的意義。 結(jié)果： 1.成功建立基于上轉(zhuǎn)發(fā)光免疫層析技術(shù)定量檢測HBV-LP的試紙條；產(chǎn)品相關(guān)性能檢測結(jié)果良好,最低檢測限約為0.1U/ml, cut-off值約為1.0U/ml,批內(nèi)與批間CV%均小于15%。 2.60例健康體檢人員血清經(jīng)UPT-LF法檢測,IHBV-LP含量均小于1U/ml,特異性為100%,ELISA法檢測結(jié)果顯示59例血清標(biāo)本HBV-LP陰性,另一例陽性,特異性為98.3%。 3.495例乙肝患者血清標(biāo)本同時(shí)采用UPT-LF法與ELISA法檢測HBV-LP,兩種方法檢測總符合率為91.7%,UPT法檢測HBV-LP陽性率為58.5%,ELISA法檢測HBV-LP陽性率為59.5%。 4.HBV-LP陽性結(jié)果濃度水平CHB組主要以40U/ml的高濃度為主,LC組和PHCC組主要集中在10U/ml的低濃度范圍內(nèi)。 5.HBeAg、HBV DNA和HBV-LP的陽性率均表現(xiàn)為CHB組LC組PHC組,HBeAg和HBV DNA在LC和PHCC患者中的陽性率均明顯低于CHB患者,差異有統(tǒng)計(jì)學(xué)意義(P0.01)。 6.HBV-LP在LC和PHCC患者中的陽性率明顯高于HBV DNA和HBeAg,差異有統(tǒng)計(jì)學(xué)意義(P0.05) 結(jié)論：上轉(zhuǎn)發(fā)光免疫層析法檢測血清HBV-LP簡便、快捷、可靠,特異性優(yōu)于ELISA法,并且HBV-LP的檢測對(duì)于慢性肝病進(jìn)展的診斷以及療效監(jiān)測有重要的價(jià)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 .
【學(xué)位授予單位】：浙江大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2015
【分類號(hào)】：R446.6

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