时间分辨荧光免疫分析法和酶联免疫吸附实验法测定乙型肝炎病毒大蛋白的方法学比较  

Methodological comparison of detecting HBV-LP by time-resolved fluoroimmunoassay and enzyme-linked immunosorbent assay

作　　者：李梅[1] 刘洁[1] 叶燕[1] 俞蕾[1] 王婷婷[1] LI Mei;LIU Jie;YE Yan(Department of Clinical Laboratory,Wuxi People′s Hospital Affiliated to Nanjing Medical University,Jiangsu 214023,China)

机构地区：[1]南京医科大学附属无锡市人民医院医学检验科,江苏214023

出　　处：《中国临床新医学》2018年第2期118-121,共4页CHINESE JOURNAL OF NEW CLINICAL MEDICINE

基　　金：无锡市医管中心科研项目(编号:YGZXM14013)

摘　　要：目的对时间分辨荧光免疫分析(time-resolved fluoroimmunoassay,TRFIA)和酶联免疫吸附实验(enzyme-linked immunosorbent assay,ELISA)检测乙型肝炎患者血清中病毒大蛋白(hepatitis B virus large surface protein,HBV-LP)进行方法学比较。方法收集30例正常体检血清标本作为阴性对照和150例慢性乙型肝炎患者血清标本,利用ELISA法检测HBV-LP,将其中70例阳性标本作为阳性样本,70例阴性标本作为阴性样本,再分别采用TRFIA法检测HBV-LP,对二者的灵敏度、特异度、一致性、检测线性范围、精密度、相关性及两种试剂盒的稳定性进行比较。其中TRFIA与ELISA结果不一致的12例标本采用PCR法进行验证。结果TRFIA检测HBV-LP的最小测定值为0.1 ng/ml,ELISA检测HBV-LP的最小测定值为2.5 ng/ml,二者的特异度均为100%。TRFIA和ELISA检测HBV-LP的Kappa值为0.83。12例ELISA和TRFIA检测不一致的标本,经PCR验证后有10例HBV DNA>103拷贝数。TRFIA试剂检测的线性范围在0.625~10 252 ng/ml之间,ELISA试剂盒检测的线性范围在5~1 281.6 ng/ml。TRFIA法检测高、中、低3个浓度水平的批内CV平均为6.10%,ELISA法的批内CV平均为8.98%。TRFIA批间CV平均为6.91%,ELISA的批间CV平均为10.45%。TRFIA试剂盒的结合下降率为6.61%,ELISA试剂盒的结合下降率为23.59%。结论 TRFIA与ELISA具有高度的一致性,但是两者相比,前者有更高的灵敏度和精密度,且TRFIA试剂盒的稳定性更好。ObjectiveTo compare the measure results of serum HBV LP between using time resolved fluoroimmunoassay(TRFIA)and enzyme linked immunosorbent assay(ELISA)in the patients with hepatitis B.MethodsA total of 150 serum specimens of hepatitis B patients were collected.30 serum specimens of healthy people were taken as the negative controls.HBV LP of 150 serum specimens was measured by ELISA to distinguish 70 negative from 70 positive samples.Then,these samples were measured by TRFIA.The sensitivity,specificity,consistency,detection range,precision,relativity and the stability of kits were evaluated.12 inconsistent samples between using ELISA and TRFIA were tested by polymerase chain reaction(PCR).ResultsThe sensitivity of TRFIA and ELISA was 0.1 ng/ml and 2.5 ng/ml respectively.The specificity of TRFIA and ELISA was 100%.The Kappa was 0.83.The HBV DNA copies of 10 inconsistent samples were>103.The detection range of TRFIA was between 0.625 ng/ml and 10.252 ng/ml.The positive rate of ELISA was between 5 ng/ml and 1 281.6 ng/ml.The intra assay average CVs of TRFIA and ELISA were 6.10%and 8.98%respectively for 3 different concentrations.The inter assay average CVs of TRFIA and ELISA were 6.91%and 10.45%respectively.The stability of the established assay kit was better than that of the commercially ELISA kit.ConclusionTRFIA and ELISA come with a high degree of consistency in detecting HBV LP.However,the sensitivity,precision and stability of TRFIA are better than those of ELISA.

关 键 词：乙型肝炎病毒 时间分辨荧光免疫分析 大蛋白 

分 类 号：R446.6[医药卫生—诊断学]