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Comparative Studies on Detection of Antibodies against Infectious Bursal Disease Virus with Test

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Agricultural Science&Technology201 2,1 3(9):1 826—1 828 ,Copyright 2012,Information Institute of HAAS.AIl rights reserved Agricultural Biotechnology Comparative Studies on Detection of Antibodies against Infectious Bursal Disease Virus with Test Strips and Agar Gel Immunodiffusion Method Jinliang ZHANG’,2 Wentong ZHANG。,Sishun HU’,Dingren BI ,Xiliang WANG’,Yuncai XIAO’ 1_College of Veterinary Medicine,Huazhong Agricultural UniversityWuhan 430070,China; ,2.College of Life Science,Shangqiu Normal Universiy,Shangqitu 476000,China; 3.Shandong Binzhou Animal Science&Veterinary Medicine Academy,Shandong Academy of Agricultural SciencesBinzhou 256600.China ,sity:agar-get-precipitin(AGP)anti- Abstract【0hiective】This study aimed to compare the detection results of antibodies veragainst infectious bursal disease virus with test strips and agar口eI Immunodifusion were detected using test strips developed in our laboratory.and the resutts were gen and AG P.positive serum of infec. chased frOm China lnstitute of Veteri— ous bursal disease virus were pur— method.[Methy1]Antibodies against infectious bursal disease virus in chicken serum ticompared with that using traditionaI agar difusion method. [Result1 The comparative study of the two methods showed that the sens ̄iviy oft test strips was eight times over agar gel immunodifusion;test strips showed higher detection rate in the deter— nary Drug Control;other chemical reagents were all made in China. Methods Strip test and result determination Test strips were used for detection at room temperature.During the de- tection,antibodies in the samples were mination test of 216 clinicaI samples,with high specfficiy,easy prteservation,and simple and rapid operation.thereby being more suitable for fhe monitoring of clinical antibodies. [Conclusion】Test strips could replace the existing serologicaI methods, having great promotion and application value in antibody monitoring.  ’Key words Infectious bursaI disease;Agar geI immunodifusion;Test strips combined with colloidaI gold—labeled chicken、IgG fFc fragment)antibodies, I nfectious bursal disease(IBD)is serum neutralizatiOn test. and en— infectious bursal disease antibodies were specifically combined with re- combinant Vp2 protein of infectious bursaI disease virus.thereby forming I an acute and highly contagious 0 chicken disease caused by jnfec. zyme-Iinked immunosorbent assay【3】. However,these methods are mainly used for laboratory test but not suitable for detection in the fields due to its re. strictions of time-consuming.high cost and requirement for expensive equip- ments.ColloidaI gold immunochro— matographic test strip was an immune tious bursal disease virus(IBDV), which directly causes certain mortality of infected chickens.and indirectly Ieads to Iong-term and severe immune suppression of subclinically infected or resistant chickens,increased suscep— red stripes at the test line.On the con- trary,the negative samples contained no infectious bursaI disease—specific antibodies,and could not form the red stripe at the test Iine.Excess colloidal tibility of chickens to other epidemics such as avian influenza and Newcastle disease and reduced ability to respond to the vaccine,thereby resulting in marker technology applying colloidal gold markers to the antigen—antibody reaction using fiber membrane as the carrier,which has only been applied in antigen detection of infectious bursal gold-labeled chicken IgG (Fc frag‘ ment)antibodies would combine with rabbit anti-mouse IgG thus forming the controlIine. immunization failure against deadly infectious diseases【”.At present.in- fectious bursal disease has been inter- nationally recognized as one of the three epidemics of chickens as a very thorny issue. Serum antibody test is one of the impo ̄ant measures to control infec- tious bursal disease.Currently.there are many l BD serum antibody detec— tion methods.including agar geIim— Specificity and sensitivity tests of lBDV antibodies using test strips disease.1n this study,self—developed test strips were used for detection of clinical samples and compared with Specificiy ttest The prepared test strips were used for detection of 0.85%saline.positive serum of infec— traditionaI AGID. tious bursaI disease virus(IBDV),pos- Materials and Methods Materials itive serum of Mycoplasma gallisep— ticum(MG),positive serum of New- Test strips for rapid detection of infectious bursaI disease antibodies castle disease virus(NDV),positive serum of Mycoplasma synoviae(MS), positive serum of infectious bronchitis were prepared in College of Veterinary munodifusion method(ACID)[a,micro- Medicine,Huazhong Agricultural Uni一 virus(IBV),positive serum of H5 avian influenza virus(H5 AIV),positive serum 0f H7 avian influenza virus(H7 AIV), positive serum of H9 avian influenza 圈 Corresponding author.E-mail:bidingren@mail.hzau.edu.cn Accepted:August 28,201 2 Received:August 4,201 2 virus (H9 AIV),and SPF chicken 1B2B 2012 antibodies were compared.According to the determination and comparison results of positive samples.the titers measured by using test strips were eight times of that measured with agar quirement for expensive experimental equipments and professionaI Iabora- tory personne1.the above methods can not meet the requirements for 究)fJ].Chinese Journal of Veterinary Science and Technology(中国兽医科 技),1 992,22(5):7-9. [3】MA XL(马 秀丽),CUI YS(崔言顺), ZHANG XM(张秀美),etaL Develop— ment and validation of indirect enzyme- Iinked immunosorbent assay test kit for rapid on-site detection[4-5].Compared wIlh these methods.the test strip de— veloped in our Iaboratory is rapid.ac- gel immunodifusion method;the test strips showed strong positive reaction with standard positive serum of infec- detection of lBDV antibody(鸡传染性法 氏囊病间接ELISA诊断试剂盒的研制 及初步应用)『J1.Chinese JournaI of curate and sensitive,and the results can be determined within 1 0 min with naked eyes.In addition,the test strip tious bursal disease virus.while no crOsS-reaCtiOn with other standard positive serum,SPF chicken serum and 0.85%saline.Test strips can be at least three months.with no signifi- cant changes ln the specificity and sensitivity.The positive derection rate of clinicaI samples using test strips reached 60.1 9%.showing a coinci- method has simple operation and can Veterinary Science(@国兽医学报),2003, 23(5):424-426. f41 WANG X,Ll K,SHl D,efa1.Develop— ment and validation of an immunochro- matographic assay for rapid detection of be used by ordinary farmers.which methods and shows great application value in antibody monitoring. stored at room temperature or 4 oC for could replace the existing serologicaI sulfadiazine in eggs and chickensfJ1.J Chromatogr B Analyt Technol Biomed Life Sci,2007,2(847):289-295 HeterenCeS 『51 PENG D,HU S,HUA Y,ef a1.Compari- son of a new gold--immunochromato-- graphic assay for the detection of anti— bodies against avian influenza virus with hemagglutination inhibition and [1】GAO YL筛玉龙),GAO HL墒宏雷 WANG XY(E.晓艳1.et aL Immunogenicity of re— combinant VP2 protein of infectious bursal disease virus expressed in re- dence rate of 68.52%with agar gel immunodifusion method.Therefore. the test strip is specific,sensitive and stable. agar gel immunodifusion assays[J].Vet- erlnary lmmunology and Immunopath・ combinant baculoviruses(鸡传染性法氏 囊病病毒VP2基因在杆状病毒表达系 Currently,agar gel immunodifu— ology,2007,1 1 7(5):1 7-25. 【6】YAN XJ,LI F,QIN LT,ef a1.Study on the knockout and the soluble prokaryot— ic expression of VP5 protein transmem- sion method (AGID),micro—serum neutralization test.and enzyme-linked immunosorbent assay are the main methods for detection of infectious bursal disease antibodies.However, due to the complex operation and re- Responsible editor:Xiaohui FAN 统中的表达及免疫原性研究)[J].Chi— nese JournaI of Preventive Veterinary Medicine(中国预防兽医学报),2007,29 (6):427—431. [2】LUO HL(罗函禄),FAN WM(范文明), ZHANG JY(张菊英),et a1.SPA COOrdi. nation agglutination test diagnosis of brane region of lBDV[J].AgricuituraI Sci- ence&Technology,201 1,12(4):621— 624. 旧D(SPA协同凝集试验诊断旧D的研 Responsible proofreader:Xiaoyan WU 试纸条法与琼脂扩散法检测IBDV抗体的比较 张进良1,2张文通3胡思顺 ,毕丁仁 ,王喜亮 ,肖运才 (1.华中农业大学动物医学院,湖北武汉430070;2.商丘师范学院生命科学学院,河 南商丘476000;3.山东省农业科学院滨州分院山东省滨州畜牧兽医研究院,山东滨州256600) 摘要[目的]比较分析IBDV抗体的试纸条与琼脂糖扩散两种检测方法。[方法]用自行研制的试纸条法检测鸡血清中的鸡传染性法氏囊病毒抗 体,并与传统的琼脂扩散法检测结果进行比较。[结果]试纸条较琼脂凝胶扩散试验敏感8倍,在216份I临床样品测定试验中,检出率更高,同时其 又具有特异性强、易保存、操作简单快速等优点,故试纸条更适合于临床抗体监测。[结论 E研究结果表明试纸条可替代现有的血清学方法,在抗 体监测方面具有极大的推广应用价值。 关键词传染性法氏囊病;琼脂扩散法;试纸条 作者简介收稿日期张进良(1980一),男,河南商丘人,博士研究生,讲师,主要从事兽医微生物学与免疫学研究,E-maihsuper20008@qq.oom。 通讯作者,教 授,硕士,博士生导师,从事兽医生物学与免疫学研究,E—mail:bidingren@mail.hzau.edu.on。 2012—08—04 修回日期2012—08-28 (Continued fr0m page 1 825) 蔽果金腰叶内寄生链格孢属真菌特性研究 段双全,许鹏辉 ,邢顺林(大学理学院,拉萨850000) 摘要[目的1了解虎耳草科植物蔽果金腰内生菌的生物学特性。[方法]利用PDA培养基分离蔽果金腰叶内的真菌,利用PDA培养基、察氏培养 基、改良型KB培养基分别对分离得到的菌株进行培养。[结果]菌株在PDA培养基、KB培养基和察氏培养基上均为菌落舒展,白色毡帽状,等径生 长,边缘整齐,呈波状,菌落到后期变为青褐色;孢子黑色,在PDA培养基上生长在菌落表面,基质呈黑色;而在KB培养基和察氏培养基上生长在 菌落边缘,基质呈黑色。孢子形态多样,多数为倒棒状,也有球形和椭圆形的,具有纵、横或斜的真隔膜,呈砖格状分隔,喙为短柱状假喙。[结论]分 离得到的菌株属于半知菌亚门丝孢纲丝孢目暗色孢科链格胞属真菌的一种。 关键词寄生;半知菌;链格胞属;孢子形态 作者简介收稿日期段双全(1962一),男,日喀则人,副教授,研究方向:微生物资源方向。 通讯作者,硕士,讲师,研究方向:植物生理生态。 2012—07—23 修回日期2012—09—03 Agricultural Science&Technology Vo1.13,No.9,2012 

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