摘要
目的建立宫颈上皮细胞中人乳头瘤病毒(HPV)16/18型E6蛋白的荧光免疫层析联合定量检测方法。方法采用荧光免疫层析原理和双抗体夹心法,制备荧光免疫层析联合检测试纸条,并对线性范围、精密性、特异性等性能指标进行评价。结果实现了对HPV16/18 E6蛋白的联合定量检测;HPV16 E6蛋白线性范围为2.0~200 ng/ml,HPV18 E6蛋白线性范围为2.0~200 ng/ml;与鳞状细胞癌抗原(SCCa)、人附睾蛋白4(HE4)无明显交叉反应;批内CV均小于10%,批间CV均小于15%,平均回收率分别为102.4%和98.1%;检测经临床病理确诊的宫颈癌样本46份、癌前病变CINⅢ样本38份及健康样本52份,本方法敏感性分别为91.3%、84.2%,特异性达到100%。结论初步建立了联合定量检测HPV16/18 E6蛋白的荧光免疫层析方法,为临床筛查宫颈癌提供一种新的检测手段。
This study aimed to establish a method of fluorescence immunochromatography for quantitative detection of human papillomavirus（HPV） 16/18 E6 protein in human cervical epithelial cells. Fluorescence immunochromatographic assay and double antibody sandwich method were used to prepare fluorescence immunoassay test strip, and the linear range, precision and specificity were evaluated. The quantitative detection of HPV16/18 E6 protein was achieved. The linear range of the method for HPV16 E6 protein detection was 2.0-200 ng/ml and the linear range for HPV18 E6 protein was 2.0-200 ng/ml. There was no significant cross-reaction with SCCa and HE4 in the detection processes. The intra-assay CV was less than 10%, the inter-assay CV was less than 15%,and the average recovery rates were 102.4% and 98.1% respectively. The sensitivity of this method were 91.3%,84.2% and the specificity up to 100% in detecting 46 cervical cancer samples, 38 precancerous lesions CIN Ⅲsamples and 52 healthy samples. In conclusion, the fluorescence immunochromatographic assay for the quantitative detection of HPV16/18 E6 protein has been preliminarily established,which provides a new detection method for clinical screening of cervical cancer.
作者
石英杰
王云龙
明亮
李玉林
王继创
潘维成
SHI Yingjie;WANG Yunlong;MING Liang;LI Yulin;WANG JichuangZ,;PAN Weicheng(Department of Clinical Laboratory, First Affiliated Hospital of Zhengzhou University, Zhengzhou 450052, Chin;Henan Bioengineering Technology Research Center, Zhengzhou 450001, China;Henan Engineering Laboratory for Biomarker Quantitative Detection, Zhengzhou 450046, China;Henan University Joint National Laboratory for Antibody Drug Engineering, Zhengzhou 4500046, China)
出处
《免疫学杂志》
CAS
CSCD
北大核心
2018年第6期534-538,546,共6页
Immunological Journal
基金
河南省科技攻关项目(162102310008)
