国际医学寄生虫病杂志
國際醫學寄生蟲病雜誌
국제의학기생충병잡지
INTERNATIONAL JOURNAL OF MEDICAL PARASITIC DISEASES
2012年
1期
5-8
,共4页
蔡玉春%郭俭%陈韶红%田利光%陈木新%艾琳%张玲玲%陈家旭
蔡玉春%郭儉%陳韶紅%田利光%陳木新%艾琳%張玲玲%陳傢旭
채옥춘%곽검%진소홍%전리광%진목신%애림%장령령%진가욱
日本血吸虫%循环抗原%IgY%间接红细胞凝集试验
日本血吸蟲%循環抗原%IgY%間接紅細胞凝集試驗
일본혈흡충%순배항원%IgY%간접홍세포응집시험
Schistosoma japonicum%Circulating antigen%IgY%Indirect hemagglutination test
目的 建立基于IgY的血吸虫循环抗原的间接红细胞凝集试验(indirect hemagglutination test,IHA)并初步探讨IgY在诊断日本血吸虫病中的应用价值.方法 以日本血吸虫卵可溶性虫抗原(soluble egg antigen,SEA)免疫海蓝蛋鸡,获得抗SEA的IgY抗体.取人O型红细胞醛化、鞣化,形成鞣化后红细胞,并分别用磷酸盐、碳酸盐和柠檬酸盐缓冲液稀释抗SEA-IgY抗体,以确定IgY致敏红细胞最佳缓冲液.将IgY溶解于最佳的缓冲液中致敏鞣化后红细胞,形成IgY红细胞检测液,用此检测液分别检测感染30、20、10条日本血吸虫尾蚴和正常小鼠血清各10份,初步检测不同感染度小鼠血清的循环抗原.结果 pH7.2磷酸盐缓冲液溶解的IgY所得致敏红细胞检测血清最佳,效价可达1∶320.用此IgY-IHA法检测中、高感染度血清(感染20条和30条日本血吸虫尾蚴)阳性率为100％,低感染度血清(感染10条日本血吸虫尾蚴)阳性率为70％,对照组10只小鼠血清全为阴性,特异性为100％.结论 建立了IgY-IHA法,此法检测日本血吸虫病循环抗原具有较高的敏感性.
目的 建立基于IgY的血吸蟲循環抗原的間接紅細胞凝集試驗(indirect hemagglutination test,IHA)併初步探討IgY在診斷日本血吸蟲病中的應用價值.方法 以日本血吸蟲卵可溶性蟲抗原(soluble egg antigen,SEA)免疫海藍蛋鷄,穫得抗SEA的IgY抗體.取人O型紅細胞醛化、鞣化,形成鞣化後紅細胞,併分彆用燐痠鹽、碳痠鹽和檸檬痠鹽緩遲液稀釋抗SEA-IgY抗體,以確定IgY緻敏紅細胞最佳緩遲液.將IgY溶解于最佳的緩遲液中緻敏鞣化後紅細胞,形成IgY紅細胞檢測液,用此檢測液分彆檢測感染30、20、10條日本血吸蟲尾蚴和正常小鼠血清各10份,初步檢測不同感染度小鼠血清的循環抗原.結果 pH7.2燐痠鹽緩遲液溶解的IgY所得緻敏紅細胞檢測血清最佳,效價可達1∶320.用此IgY-IHA法檢測中、高感染度血清(感染20條和30條日本血吸蟲尾蚴)暘性率為100％,低感染度血清(感染10條日本血吸蟲尾蚴)暘性率為70％,對照組10隻小鼠血清全為陰性,特異性為100％.結論 建立瞭IgY-IHA法,此法檢測日本血吸蟲病循環抗原具有較高的敏感性.
목적 건립기우IgY적혈흡충순배항원적간접홍세포응집시험(indirect hemagglutination test,IHA)병초보탐토IgY재진단일본혈흡충병중적응용개치.방법 이일본혈흡충란가용성충항원(soluble egg antigen,SEA)면역해람단계,획득항SEA적IgY항체.취인O형홍세포철화、유화,형성유화후홍세포,병분별용린산염、탄산염화저몽산염완충액희석항SEA-IgY항체,이학정IgY치민홍세포최가완충액.장IgY용해우최가적완충액중치민유화후홍세포,형성IgY홍세포검측액,용차검측액분별검측감염30、20、10조일본혈흡충미유화정상소서혈청각10빈,초보검측불동감염도소서혈청적순배항원.결과 pH7.2린산염완충액용해적IgY소득치민홍세포검측혈청최가,효개가체1∶320.용차IgY-IHA법검측중、고감염도혈청(감염20조화30조일본혈흡충미유)양성솔위100％,저감염도혈청(감염10조일본혈흡충미유)양성솔위70％,대조조10지소서혈청전위음성,특이성위100％.결론 건립료IgY-IHA법,차법검측일본혈흡충병순배항원구유교고적민감성.
Objective To establish anti-SEA-IgY based indirect hemagglutination test (IHA) method of detecting circulating antigen of Schistosoma japonicum and explore its application value for diagnosis of schistosomiasis.Methods Hens were immunized with soluble egg antigen (SEA) of S.japonicum,specific anti-SEA IgY was purified from eggs after immunization.The anti-SEA IgY was diluted with three different buffer as phosphate (PBS,pH7.2),carbonate ( pH 9.6) and citrate buffer to choose the best buffer.After aldehyding and tanning,the human O type erythrocytes were sensitized with the anti-SEA IgY which was dissolved into the best buffer,formatting the IgY adsorbed erythrocytes test solution.With this test solution,30 sera of mice infected with 10,20 or 30 S.japonicum cercariae respectively and 10 sera of normal control mice were tested for detecting circulating antigens in the sera of mice infected with S.japonicum. Results Erythrocytes which were sensitized with the anti-SEA IgY dissolving into phosphate buffer (pH7.2) gave the best results,the best titer was up to 1:320.With this IgY-IHA method,the positive rate of sera of high degree infection( mice infected with 20 and 30 cercariae) were 100％,while the positive rate of sera of low degree infection(mice infected with 10 cercariae) were 70％,10 sera of control group mice were all negative.Conclusion The established IgY-IHA method exhibited higher specificity andsensitivity in detecting circulating antigen of S.japonicum.
