CAJ | 학술논문

目的研制快速诊断广州管圆线虫感染的金标层析检测试剂.方法双抗体(12D5和21B7)夹心ELISA检测实验感染广州管圆线虫的大鼠、广州管圆线虫感染病例血清循环抗原(CAg),同时将12D5和21B7单抗点样于固相的硝酸纤维膜上,以胶体金-protein A为标记物制备金标免疫层析试剂快速检测实验感染广州管圆线虫的大鼠、广州管圆线虫感染病例血清循环抗原.结果经鉴定单抗12D5为IgG1,21B7为IgM,两株单抗同时识别广州管圆线虫成虫相对分子质量55×10~3的蛋白,12D5和21B7双抗体夹心ELISA和金标层析法对实验感染广州管圆线虫的大鼠血清中CAg检出率为100%(48/48),广州管圆线虫感染病例血清CAg检出率为100%(32/32),与日本血吸虫、肝吸虫、肺吸虫、旋毛虫、蛔虫、包虫病例血清无交叉反应,与健康人血清无反应.结论 12D5、21B7双抗体夹心ELISA和金标层析法对感染广州管圆线虫人和动物血清中CAg检测的特异性强,金标层析法操作简便、快速,结果判读容易,不需特殊设备,且敏感性高,能够确定现症感染.
목적 연제쾌속진단엄주관원선충감염적금표층석검측시제.방법 쌍항체(12D5화21B7)협심ELISA검측실험감염엄주관원선충적대서、엄주관원선충감염병례혈청순배항원(CAg),동시장12D5화21B7단항점양우고상적초산섬유막상,이효체금-protein A위표기물제비금표면역층석시제쾌속검측실험감염엄주관원선충적대서、엄주관원선충감염병례혈청순배항원.결과경감정단항12D5위IgG1,21B7위IgM,량주단항동시식별엄주관원선충성충상대분자질량55×10~3적단백,12D5화21B7쌍항체협심ELISA화금표층석법대실험감염엄주관원선충적대서혈청중CAg검출솔위100%(48/48),엄주관원선충감염병례혈청CAg검출솔위100%(32/32),여일본혈흡충、간흡충、폐흡충、선모충、회충、포충병례혈청무교차반응,여건강인혈청무반응.결론 12D5、21B7쌍항체협심ELISA화금표층석법대감염엄주관원선충인화동물혈청중CAg검측적특이성강,금표층석법조작간편、쾌속,결과판독용역,불수특수설비,차민감성고,능구학정현증감염.
Objective To detect the rate of Angiostrongylus cantonensis infection and to study the effects of treatment so as to prepare monoclonal antibodies (McAbs) ,and gold immunochroma-tography assay (GICA) with 12D5 and 21B7 McAbs could be prepared in advance. Methods Two McAbs (12D5 and 21B7) were applied to detect the circulating antigen (CAg) in the sera of rats infected with A. cantonensis and angiostrongyliasis patients respectively by double antibody sandwich ELISA. Either 12D5 or 21B7 McAbs was used as antibody and protein A was conjugated with colloid gold as the detection marker. A special pad for GICA was designed according to the reaction procedure, and CAg were detected by GICA in the sera of rats infected with A. cantonensis and angiostrongyliasis patients respectively. Results 12D5 McAb was identified as IgG1 and 21B7 McAb was IgM. Results from Western blotting showed that two McAbs could be used to identified 55 KD protein of adult worms of A. cantonensis. The detection rates of CAg in the sera of infected rats was 100% (48/48) and the detection rates of CAg in the sera of angiostrongyliasis patients was 100% (32/32). No cross-reaction to sera of patients with other infection of parasites, such as clonochiasis, fasiolopsiasis, ancylostotniasis, ancylostomiasis, anisakiasis as well as schsitosomiasis wee seen and normal sera did not react with 12D5 and 21B7 McAbs. Conclusion Results from sandwich ELISA and GICA with 12D5 and 21B7 McAbs showed high specificity and acting as detecting CAg of A. cantonensis in sera of infected animals and patients. We noticed that GICA with 12D5 and 21B7 was not only rapid and simple that without requirement of special instrument, but also rather sensitive and specific for the detection of current infection with A.cantonensis.