金免疫层析法检测囊虫循环抗原试验研究  

The Detection of the Circulating Antigen(CA) of Taenia solium Gysticercosis with Gold Immunochromatographic Assay (GIGA)

作　　者：唐雨德[1] 周东明[1] 

机构地区：[1]南京军区军事医学研究所,江苏南京210002

出　　处：《动物医学进展》2004年第5期87-88,共2页Progress In Veterinary Medicine

摘　　要：应用胶体金和免疫层析技术进行了检测金免疫层析法(GICA)囊虫循环抗原(CA)试验研究。5株抗囊虫McAb和一株多抗用于最佳配对的筛选,一株用于标记胶体金,另一株包被NC膜。以双抗体夹心ELISA为对照,GICA用于CA检测,观察其敏感性、特异性和稳定性。结果显示,以McAb 1A5和1B6配对的结果最佳,1A5最适标记量为10μg／mL,186最佳包被浓度为1 g／L;用GICA和ELISA两种方法检测囊虫CA,痈猪血清和囊液的符合率达100％,病人血清和脑脊液的符合率达80％;CA最低检测量为10 ng,检测正常猪、人及其他疾病血清,均呈阴性;检测时间5-15 min,整个实验仅一步操作;GICA试纸条分别在4℃和室温下保存105 d以及在37℃保存45 d,检测结果均未发现改变。以上结果表明,快速检测囊虫循环抗原的金免疫层析法具有简便、快速、敏感、特异和稳定的特点,适合基层使用。For developing a simple, fast, easy-to-do and new method to detect the circulating antigen(CA) of Taenia solium cysticercosis for county veterinarian, a gold immunochromatographic assay (GICA) was established by combining colloidal gold technique with immunochromatographic assay. Five McAbs and one PcAb a gainst CA were used for screening the optimal composition, and two McAbs were purified and applied for preparation of GICA strips by labeling one McAb with colloidal gold and coating another on nitrocellulose (NC) membrane. As compared with sandwich ELISA, when GICA was used to detect CA, its sensitivity, specificity and stability were evaluated. The optimal composition was McAb 1A5 and 1B6. The optimal marked amount of 1A5 with one-milliliter colloidal gold was 10 μg/mL,and the optimal coating concentration of 1B6 was 1 mg/ mL. Concordance between GICA and sandwich ELISA for detecting CA of the sera of sick pigs and cyst fluid was 100%,and 80 % for the sera and cerebrospinal fluid of patients. Threshold value of GICA was 10ng of CA. The results were negative for detecting the sera of normal pigs, human and other disease pigs. The whole detecting time was 5 to 15 min. GICA strips coated with Abs can be stored at 4 ℃ and room temperature for 105 d and 37 ℃ for 30 d,repectively. It concluded that GICA detecting CA was a suitable method because of its convenience, rapidity, sensitivity, specificity and stability.

关 键 词：金免疫层析法 检测 囊虫病 循环抗原 

分 类 号：S855.91[农业科学—临床兽医学] S852.4[农业科学—兽医学]