摘要
目的 建立一种快速检测NJ001特异性抗原的胶体金免疫层析法.方法 以肺腺癌细胞株SPC-A1为抗原免疫新西兰兔,制备多抗.在硝酸纤维素膜上分别包被多抗和羊抗鼠IgG,作为检测线和质控线,用胶体金标记单抗N J001滴加在玻璃纤维素膜上,组装成胶体金免疫层析试纸条检测NJ001特异性抗原.用上述制备的试纸条对2007年10月至2013年11月从南京医科大学第一附属医院收集的经病理确诊的52例肺腺癌患者（肺腺癌组）和40名健康体检者（健康对照组）血清中NJ001特异性抗原进行检测.肺腺癌组中有TNM分期资料的NSCLC患者49例,早期组（Ⅰ/Ⅱ期）15例,晚期组（Ⅲ/Ⅳ期）34例,3例无TNM分期资料;45例有淋巴转移资料的患者中无淋巴结转移者22例,有淋巴结转移者23例,7例无淋巴结转移资料.组间阳性率比较采用x2检验或Fisher确切概率法.结果 在52例肺腺癌患者血清中N J001特异性抗原的阳性率为55.77％,高于40名健康体检组阳性率（7.5％,x2=23.22,P＜0.001）.49例有TNM分期资料的肺腺癌患者中34例Ⅲ/Ⅳ期肺腺癌患者血清的NJ001特异性抗原的阳性率高于15例Ⅰ/Ⅱ期者（64.17％ ＞33.33％,x2=4.141,P =0.042）.45例有淋巴转移资料的患者中23例有淋巴结转移的患者阳性率高于22例无淋巴结转移者（69.57％ ＞36.36％,x2=4.98,P=0.026）.结论 本研究成功建立了快速检测NJ001特异性抗原的胶体金免疫层析法,可以用于临床血清样本的检测.
Objective To develop a colloidal gold immunochromatographic assay for rapid detection of NJ001 specific antigen. Methods Polyclonal antibody was prepared by immunizing New Zealand rabbit with SPC-Al cells. Polyclonal antibody and Goat anti mouse IgG, as test line and control line respectively, were immobilized on nitrocellulose membrane, NJ001 was labeled by colloidal gold particles and then sprayed and fixed on fiberglass membrane as the combination pad. Together with a specimen pad ahead. The nitrocellulose membrane and the eonbination pad were assembled into a test strip. The poisitive rate of NJ001 were examined from 52 lung adenoearcinoma patients and 40 healthy controls （ HC ） . In the lung adenocarcinoma group, 49 were comfirmed by TNM, 15 stage Ⅰ/Ⅱ 34 stage Ⅲ/Ⅳ, and 3 cases without TNM staging data. 45 patients had lymphatic metastasis data, 22 cases were of no lymphatic metastasis and 23 cases were of lymphatic metastasis. There were another 7 cases without the data of lymphatic metastasis. Results Positive rate of NJ001 in lung adenocarcinoma patients was 55.77% , significantlyhigher than that of HC （7.5 % ） （X2 = 23.22, P 〈 0. 001 ）. Positive rate of N J001 specific antigen in patients at TNM stage Ⅲ/Ⅳ was higher than that at TNM stage Ⅰ/Ⅱ （64. 17% vs 33.33% ,x2 =4. 141,P = 0. 042 ） ; Positive rate in patients with lymph nodes metastasis was higher than that in patients without lymph nodes metastasis （ 69.57% vs 36. 36% , X2 = 4. 98, P = 0. 026 ） . Conclusion This study successfully establishes a colloidal gold immunochromatographic assay that can be used to rapid detect NJ001 specific antigen in clinical serum samples.
出处
《中华检验医学杂志》
CAS
CSCD
北大核心
2014年第7期548-552,共5页
Chinese Journal of Laboratory Medicine
基金
国家自然科学基金资助项目（81371894、81272324、81000754、81101322、81201359）
国家临床检验重点专科建设项目基金资助
江苏高校优势学科建设工程基金项目资助课题
江苏省实验诊断学重点实验室基金资助项目（XK 201114）
教育部博导基金（20113234110012）
关键词
肺肿瘤
腺癌
抗体
单克隆
胶体金
免疫层析法
Lung neoplasms
Adenocarcinoma
Antibodies, monoclonal
Gold colloid
Immunochromatography
