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1.
以皱纹盘鲍脓足病致病病原-创伤弧菌为抗原,制备兔抗血清,抗体纯化后以辣根过氧化物酶标记,建立检测创伤瓜菌的双抗夹心ELSIA检测法。结果表明,双抗夹心ELSIA法有较高的灵敏度,可检出含菌10^4个/ml的菌悬液。  相似文献   

2.
应用Dot_ELISA技术检测草鱼出血病病毒的研究   总被引:4,自引:0,他引:4  
报道了斑点免疫吸附ELISA(Dot-ELISA)快速检测草鱼出血病病毒(GCHV)的方法以及对提纯病毒、染毒细胞内病毒和病鱼组织的检测结果,并对该方法的灵敏度与SPA-CoA和常规ELISA法进行了系统的比较。结果表明,Dot-ELISA检出GCHV的最低含量为3pg,该灵敏度比SPA-CoA和常规ELISA法分别提高10倍和20倍,而且快速易行,是目前检测GCHV最为有效的方法,并可在草鱼出血病临床诊断和病毒疫苗质量鉴定等方面得到应用。  相似文献   

3.
应用Dot—ELISA技术检测草鱼出血病病毒的研究   总被引:5,自引:2,他引:3  
报道了斑点免疫吸附ELISA快速检测草鱼出血病病毒的方法以及提纯病毒、染毒细胞内病毒和病鱼组织的检测结果,并对该方法的灵敏度与SPA-CoA和常规ELISA法进行了系统的比较。  相似文献   

4.
副溶血弧菌的外膜蛋白及其抗原性研究↑(*)   总被引:11,自引:1,他引:10  
本文对13株不同来源的副溶血弧菌的外膜蛋白及其抗原性进行了比较研究。13株副溶血弧菌的外膜蛋白有相似的SDS-PAGE图谱。大多数菌株有五条主要的蛋白质,其大致分子量分别为:a、69kDa;b、63kDa;c、40kDa;d、30kDa;e、28kDa。其中蛋白质b是所有菌株共有的。与吸附后的兔抗副溶血弧菌血清Western印迹法结果显示抗血清与多数副溶血弧菌菌株的外膜蛋白有一条共同的免疫反应带,其大致分子量为44kDa。  相似文献   

5.
近几年中国南方杂色鲍(Haliotis diversicolorReeve)养殖场频繁爆发严重的肌肉萎缩症,该病能感染各种规格杂色鲍,死亡率很高。病鲍症状主要表现为外套膜和内脏团萎缩、无生气,附着力下降,最终死亡。该试验从病鲍组织分离到1株优势菌WS,人工感染试验证实其对健康杂色鲍有很强致病性,且与自然发病的杂色鲍症状相同。对菌株WS进行形态学观察和生理生化检测,结果显示与哈维氏弧菌(Vibrio harveyi)最为接近。为了进一步确定其分类地位,采用1对通用引物扩增其16S rDNA序列片断进行序列分析和同源性对比,构建系统发育树,结果表明,菌株WS与弧菌属的V.harveyi[AY750577]聚为1个分支,且同源性达99.70%。综合上述3种鉴定结果,确定该菌为哈维氏弧菌。  相似文献   

6.
应用葡萄球菌A蛋白协同凝集试验快速检测3株鱼类致病菌   总被引:5,自引:0,他引:5  
本试验用富含葡萄球菌A蛋白的国际标准菌株cowanI分别标记3株引起民鱼类致病的荧光假单胞菌、鳗弧菌和鲁克氏耶尔森菌的特异高兔血清制成SPA诊断液,建立了检测估摸力的协仙凝集试验。将待检病料增减16-18h用该法检测3min可判定结果,比常规玻片敏感25-50倍;经交叉试验和重复性试验,表明特异性强,敏感性高可重复性好,一具有简便快速经济等特点。适用于基层鱼场,可用于淡水鱼类病原菌的快速鉴定,疫情  相似文献   

7.
皱纹盘鲍幼鲍溃烂病病原菌的ELISA检测法   总被引:8,自引:0,他引:8       下载免费PDF全文
皱纹盘鲍幼鲍溃烂病病原菌的ELISA检测法Aelisamethodfordetectingthepathogenicbacteriaoffesterdiseaseinculturedjuvenileabalone叶林俞开康王如才(青岛海洋大学水产学院...  相似文献   

8.
杂色鲍幼苗“急性死亡脱落症”病原菌分析   总被引:2,自引:0,他引:2       下载免费PDF全文
从“急性死亡脱落症”发病期的杂色鲍幼苗体内和病苗附着的薄膜上,共分离到4株优势菌,即NA0301、NA0302、NA0303和NA0304。经人工浸泡感染实验证明,从病幼苗体内和苗附着的薄膜上分离到的优势菌株NA0301和NA0302是杂色鲍幼苗的致病菌,它们均为革兰氏阴性、短杆状、极生鞭毛,能运动。常规生理生化和Biolog细菌鉴定系统测试结果表明,菌株NA0301与溶珊瑚弧菌(Vibrio coralliilyticus)的亲缘关系最近,而菌株NA0302与溶藻弧菌(Vibrio alginolyticus)的亲缘关系最近。为了进一步确定它们的分类学地位,分别测定了溶珊瑚弧菌和溶藻弧菌的16S rDNA序列,分析相关细菌相应序列的同源性,并构建系统进化树。结果表明,菌株NA0301与溶珊瑚弧菌的亲缘关系最近,菌株NA0302与溶藻弧菌的亲缘关系最近。因此,菌株NA0301和菌株NA0302分别鉴定为:溶珊瑚弧菌和溶藻弧菌。[中国水产科学,2006,13(4):655—661]  相似文献   

9.
对虾病原菌2—5B菌株16SrRNA基因片段的克隆和序列测定   总被引:2,自引:0,他引:2  
用引物PL1-PL2PCR扩增对虾病原菌-坎普氏弧菌2-5B菌株16SrRNA基因-1223bp的片段,采用pUC19质粒构建dT载体法完成该片段的克隆。部分序列测定及分析结果表明,该菌株与GenBank中坎普氏弧菌标准株序列之间同源性为96.94%。  相似文献   

10.
杂色鲍养殖环境中致病性弧菌分布调查   总被引:3,自引:0,他引:3  
从深圳不同的杂色鲍人工养殖场采集并分类环境生物样品。应用TCBS平板分离并作生理生化初步鉴定弧菌,平板涂布法统计水样和各类生物每mL(或每g)所含弧菌总数,并应用基于16S~23SrDNA间区序列设计的4种水产病原弧菌特异性引物进行PCR,定性检测各类环境样品所携带致病性弧菌的分布状况。结果显示,弧菌广泛存在于杂色鲍养殖环境中,且杂色鲍养殖池池水中的弧菌密度大于进水口海水。在养殖环境生物中,不同环境生物中每克生物所携带的弧菌数差别很大,其中盘管虫、海蟑螂、等足类所携带的弧菌数最多,而海鞘携带的弧菌数最少。在常见的4种致病性弧菌的检测结果上,创伤弧菌和溶藻弧菌阳性率均为3.4%。通过研究弧菌在杂色鲍养殖环境中弧菌的分布特征,为杂色鲍养殖中流行性弧菌病的预防提供科学依据。  相似文献   

11.
为了从对虾体内分离出的菌株中快速筛选出蜡样芽孢杆菌和溶藻胶弧菌,将蜡样芽孢杆菌与溶藻胶弧菌作为抗原,免疫SPF新西兰大白兔,获得免疫血清,效价均高于1:2000。将免疫兔血清作为一抗,HRP-羊抗兔血清作为二抗,建立了蜡样芽孢杆菌和溶藻胶弧菌快速检测的间接ELISA方法。此方法中,兔抗血清最佳稀释度为1:10000,菌液最佳包被浓度为106 CFU/ml;HRP-羊抗兔血清最佳稀释浓度为1:1000,可检测细菌最低浓度为104 CFU/ml。抗蜡样芽孢杆菌血清与苏云金芽孢杆菌有交叉反应,抗溶藻胶弧菌血清与其亲缘相近菌株无交叉反应,具有较强的特异性。2012年和2013年,分别从斑节对虾、中国对虾、凡纳滨对虾等9批样本中分离纯化到109株海洋细菌,利用建立的多抗间接ELISA方法对其中部分菌株进行快速检测,共检测出6株溶藻胶弧菌,未检测出蜡样芽孢杆菌。  相似文献   

12.
大鳞大马哈鱼生长激素抗体的制备与鉴定   总被引:1,自引:0,他引:1  
本文用大鳞大马哈鱼生长激素(sGH)免疫日本大耳白兔制备了特异抗血清。应用常规酶联免疫吸附测定法(ELISA)对抗血清效价和特异性进行了测定,结果表明sGH抗血清只与sGH特异反应,与草鱼GH、草鱼催乳素(PRL)、牛GH及大群大马哈鱼促性腺激素(sGtH)等基本上没有交叉反应,该抗血清的效价约为1:70000左右。应用sGH抗血清,初步建立了sGHELISA检测方法。  相似文献   

13.
牙鲆迟钝爱德华氏菌血清型及荧光抗体检验   总被引:3,自引:0,他引:3  
以迟钝爱德华氏菌的代表菌株(HC010907-1)为免疫原,制备免疫血清,对分离于牙鲆鱼的130株迟钝爱德华氏菌进行了血清型检定,结果表明:供试的130株迟钝爱德华氏菌均为同种血清型;同时以此免疫血清为第一抗体,以标准羊抗兔IgG荧光抗体为第二抗体,进行了荧光抗体技术检验迟钝爱德华氏菌的可行性试验,表明亦具有较强的特异性。  相似文献   

14.
Abstract A 63-year-old man was admitted to hospital for septicaemia and severe metastatic skin infection, 24h after he had eviscerated fresh eels. V. vulnificus was isolated from his blood and wounds. The strain was indole negative, was ornithine decarboxylase positive and grew at 42°C. A strain of V. vulnificus with these characteristics was isolated in 1987 from diseased eels. The characteristics differed from those of V. vulnificus strains (biogroup 1) that have been reported from patients world-wide. V. vulnificus biogroup 1 was isolated from 3 of 11 seawater samples collected along the coast of the Netherlands, but indole negative strains of V. vulnificus were not isolated. We conclude that an indole-negative variant of V. vulnificus is pathogenic for eels and for human beings and that eels may transmit V. vulnificus to humans.  相似文献   

15.
用十二烷基肌氨酸钠(Sarkosyl)抽提结合超速离心的方法提取了一株大菱鲆致病性溶藻弧菌(Vibrio alginolyticus)SR1和其他7株弧菌的外膜蛋白。通过SDS-PAGE图谱分析比较了这8株弧菌外膜蛋白的组成,结果表明,8株弧菌的外膜蛋白电泳一般可得到6-12条条带,其分子量多集中在65-106 kD和28-48 kD,其中36 kD的蛋白带为8株弧菌所共有。用兔抗SR1全菌血清进行Western-blot印迹显示,菌株SR1的外膜蛋白条带中有6条发生了阳性反应,其分子量分别为73 kD、48 kD4、5 kD3、9 kD、36 kD和32 kD。而其他7株弧菌的外膜蛋白与兔抗SR1血清也发生程度不等的阳性反应,这些阳性反应条带的分子量集中在65-73 kD、45-48 kD和36-41 kD之间,其中36 kD的外膜蛋白在8株弧菌中均出现明显的阳性反应,说明36 kD的外膜蛋白是这8株弧菌共有的特异性抗原。  相似文献   

16.
Abstract. Specificities of polyclonal salmon antisera made against the fish pathogens Vibrio salmonicida and Vibrio anguillarum were studied. Using ELISA and Western blot techniques, antisera made against V. salmonicida or V. anguillarum serovar 1 demonstrated high responses against the homologous bacterium or its isolated LPS. In contrast, antisera obtained after immunization with V. anguillarum serovar 2 displayed low antibody titres against homologous antigens. Elcctrophoretic transfer of SDS-PAGE separated V. salmonicida LPS antigen to nitrocellulose strips and subsequent immunostaining with salmon antisera revealed a strong reaction exclusively in the low molecular weight region (<14kD). On the other hand, immunoblots of V. anguillarum LPS preparations using salmon immunesera raised against this species showed a heterogenous staining pattern ranging from high to medium LPS-size. In addition, most of the salmon antisera made against V. anguillarum serovar 2 also reacted with a low molecular weight LPS antigen band.  相似文献   

17.
创伤弧菌、溶藻弧菌外膜蛋白特性的比较研究   总被引:1,自引:0,他引:1  
对用Sarkosyl法分离的创伤孤菌、溶藻弧菌、副溶血弧菌、鳗弧菌的外膜蛋白进行了初步的比较分析.这4种弧菌外膜蛋白的SDS-PAGE和Western blotting的图谱有相似性亦有差异.SDS-PAGE显示,4种弧菌中除副溶血弧菌外均能分离到47、38 ku的外膜蛋白;创伤弧菌和溶藻弧菌存在4种共同的外膜蛋白,大...  相似文献   

18.
IPTG诱导含有鲮(Cirrhinus molitorella)生长激素基因(growth hormone,GH)的工程菌M15(pQE30-GH)重组表达鲮生长激素蛋白。SDS-PAGE凝胶电泳结果表明,在24.5kDa处有1条重组表达鲮生长激素条带,经分离纯化、复性和浓缩后得到有活性的重组鲮生长激素蛋白。以复性浓缩后的重组鲮生长激素蛋白为抗原,采用改进的方法对新西兰大白兔进行皮下多点免疫注射,获得兔抗鲮生长激素多克隆抗体。经ELISA检测该免疫血清的滴度为12800。以该多抗为一抗,Western blotting可以检测到10ng的抗原量;并且在鲮垂体组织提取液中和血清中检测到一种能与该血清作用的大小约为21.5kDa的蛋白条带。说明该试验所制备的兔抗鲮生长激素多克隆抗体具有较好的免疫活性。  相似文献   

19.
Abstract. Seven marine Vibrio strains pathogenic for fish or shellfish (including strains of Vibrio alginolyticus, V. anguillarum, V. tubiashi and V. ordalii ) were categorized into three groups on the basis of their extracellular proteinases. Antiserum raised against purified Vibrio alginolyticus NCMB 1339 proteinase neutralized the enzymes produced by all seven Vibrio strains and, on immunodiffusion, the V. alginolyticus proteinase gave a reaction of partial antigenic identity with the other Vibrio strains. In experimental infections of Ostrea edulis larvae with V. alginolyticus , the production of proteinase was demonstrated by immunofluorescence with fluorescein-labelled anti-serum. The toxicity of purified proteinase to larval O. edulis was significantly reduced by preincubation with antiserum but, when larvae were challenged with V. alginolyticus culture supernate containing a similar proteinase concentration, preincubation with antiserum had no protective effect.  相似文献   

20.
Japanese flounder, Paralichthys olivaceus (Temminck et Schlegel), were immunized by the intraperitoneal (i. p.) injection route with formalin‐killed whole cells of Vibrio anguillarum that originated from a diseased fish. Fifty days later, a booster vaccination was given by the same route. Control fish were similarly treated with sterile phosphate‐buffered saline. The efficacy of vaccination was evaluated based on protection against two bacterial challenges and immune responses (both specific and non‐specific). The challenges were performed by i. p. injection with V. anguillarum or V. parahaemolyticus. The results indicated that the vaccinated fish showed higher non‐specific immune activity than the unvaccinated fish. The effects of vaccinations on the phagocytic activity of phagocyte, bactericidal and lysozyme activities were notable, especially on bactericidal activity. Determined by ELISA, antiserum of vaccinated fish displayed high antibody titres. The vaccination conferred protection against V. anguillarum challenge (81.25–93.75% relative percentage survival (RPS)). The RPS was 46.15–53.85% against V. parahaemolyticus challenge, indicating some degree of cross‐protective immunity.  相似文献   

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