牛腹腔丝虫抗原血清学免疫反应性的探讨

马、绵羊和山羊脑脊髓丝虫病在我国诸多省份常有发生。该病按其临床症状难以诊断,故常以死后剖检在脑或脊髓中发现童虫而确诊。为了寻求家畜脑脊髓丝虫病的血清学诊断方法,本文就牛腹腔丝虫成虫抗原与马、骡、山羊、黄牛和免疫家兔等动物血清的免疫反应性作了试验,探讨了牛腹腔丝虫成虫抗原用于家畜脑脊髓丝虫病血清学检测的可行性。     

羊脑脊髓丝虫病的防治

羊脑脊髓丝虫病是由寄生于牛腹腔的指形丝状线虫和唇乳突丝状线虫的幼虫迷路移行后,童虫寄生于幼羊的脑脊髓,而引起的以脑脊髓炎和脑脊髓实质破坏为特征的寄生虫病。1流行情况羊脑脊髓丝虫病有明显的季节性,多发生于夏末秋初,一般为7~10月份,其发病率随气温的升高及雨季的来临逐渐升高[1]。羊脑脊髓丝虫病在师宗县呈区域性零星散发,海拔在1 200 m以上的丹凤、雄壁、葵山等6个乡(镇)发病较少,在海拔低于1 200 m的高良、五龙2个乡(镇)发病较多,随海拔升高,发病数量逐步下降。自1999年以来,师宗县的羊因感染脑脊髓丝虫病而发病4 520只,死亡2 17…     

东北梅花鹿肾虫病的防治方法

笔者首次报道了东北梅花鹿肾虫病的发生情况、流行特点、临床症状、病理变化,以及病原体的形态特征、致病作用和本病的主要防治措施,概述了本病和鹿脑脊髓丝虫病主要症状之区别。     

羊脑脊髓丝虫病、牛副丝虫病的诊疗

<正>1羊脑脊髓丝虫病羊脑脊髓丝虫病是由寄生于羊腹腔的指形丝状线虫和唇乳突丝状线虫的幼虫迷路移行后,童虫寄生于羊的脑脊髓而引起的以脑脊髓炎和脑脊髓实质破坏为特征的疾病,又称摆腰病、趔腰病。1.1病原特点本病的病原是指形丝状线虫的晚期幼虫(童虫)。为乳白色小线虫,其形态已基本近似成虫。当幼虫侵入血管,随血流循环侵入脑脊髓内引发本病。指形线状线虫的中间宿主是蚊子,是本病的唯一传播者,本病的流行与蚊虫     

鹿脑脊髓丝虫病

自 1 98 0年以来 ,双辽市鹿场所饲养的马鹿和东北梅花鹿先后不断发生脑脊髓丝虫病。鹿患本病在1 995年以前无人报道。在本病未被人们了解时 ,饲养员们依据患鹿的临床表现称之为“偏胯病”、“拖拉胯病”、“扭腚病”、“晃腰病”、“腰痿病”或“后躯麻痹病”。从 1 990— 1 99     

唇乳突丝虫感染蚴接种非固有宿主动物致发脑脊髓丝虫病研究

用东乡伊蚊叮吮含有唇乳突丝虫微丝蚴的模型动物——小鼠血液,待微丝蚴在蚊体内发育至感染期时,分离出感染坳,皮下多点接种43只昆明小鼠(每只15～200条)、1只山羊及2只绵羊(每只200～300条)和4匹驹(每匹750～1 250条);将指状丝虫接种到20只昆明鼠体内,比较两种虫体的致病性。结果:唇乳突丝虫感染鼠有16只经1～8 d潜伏期发病,呈现瘫痪、昏迷症状后死亡.实验羊和驹经5～21d潜伏期后均呈不同程度的运动和神经症状,于接种虫体后40～140 d扑杀.经病理学观察,3种动物脑脊髓均呈现虫伤性液化坏死灶及非化脓性脑脊髓炎变化,在其中枢神经系统组织切片中发现丝虫虫体断面或钙化碎片,从而证明唇乳突丝虫可人工感染昆明小鼠、羊、驹发生脑脊髓丝虫病,进而提出该虫可以成为马、羊脑脊髓丝虫病的病原之一。指状丝虫感染鼠死亡率高于唇乳突丝虫感染鼠(P<0.01),初步认为唇乳突丝虫致病性较指状丝虫弱。     

马脑脊髓丝虫病的研究进展

<正> 马脑脊髓丝虫病(EquineCerebrospinal setariasis)是由牛腹腔丝虫的幼虫侵入马脑脊髓而引起的一寄生虫病。本病的病原学、流行病学、病理学、临床症状、诊断、治疗、免疫、预防等方面的内容在《马脑脊髓丝虫病》的专著中均作了一定程度的介绍。近年来,对本病的病原学、流行病学、诊断学、发病机制及免疫等方面又作了进一步的探讨,并取得了一些成果。现根据笔者所在的马脑脊髓丝虫病发病机制研究组实验情况和所能搜集到的资料,就《马脑脊髓丝虫病》     

绵羊脑脊髓丝虫病的诊断与防治

2017年9月-11月,新疆某县某牧场2个羊群中先后有14只羊出现后肢无力,起立困难,甚至瘫痪,3d~5d后死亡。根据临床症状、解剖后观察病理变化,确诊为绵羊脑脊髓丝虫病。本文主要介绍绵羊脑脊髓丝虫病的诊断与防治方法。     

犬心丝虫病的诊治

犬心丝虫病又称犬恶丝虫病 ,是由线虫科的犬恶丝虫寄生于犬心脏的右心室及肺动脉 (少见胸腔、支气管 )引起的循环障碍 ,呼吸困难及贫血等症状 ,除犬外 ,猫和其它野生肉食动物均可成为犬恶丝虫的终未宿主。犬心丝虫在我国分布很广 ,北至沈阳 ,南至广州均有发现。在中西部正日益成为问题 ,感染率为2 0 %～ 40 % ,高的流行率总是与蚊子密度相关连。1　病原犬心丝虫呈黄白色细长粉丝状 ,雄虫长 1 2～ 1 8ｃｍ ,尾部数回盘转。雌虫长 2 5～ 3 0ｃｍ ,尾部直。受精卵在雌虫的子宫内发育和孵化。早熟的活动胚胎称为微丝蚴 ,长 3 0 7～ 3 2 2ｍｍ。…     

用EMA-1基因重组蛋白对马巴贝斯虫病的血清学调查

在寄生虫病的血清学诊断中 ,常用的虫体抗原有纯化难、制备有限和部分交叉反应等缺点。以重组蛋白作为诊断抗原具有特异性强、敏感性高、制备简单和易纯化等优点。本试验以EMA-1基因重组蛋白为诊断抗原 ,用 EL ISA方法对延边地区马巴贝斯虫病进行了血清学调查。结果表明 ,该地区马巴贝斯虫病的血清阳性率为 3 4.2 % ( 3 8/ 1 1 1 ) ,而血涂片染色镜检法的阳性率为 1 3 .5% ( 1 5/ 1 1 1 ) ,两者之间差异极显著 ( P <0 .0 1 )。不同牧地和不同年龄被检血清阳性率之间未见显著差异。     

Evaluation of coproantigen diagnosis for natural Echinococcus multilocularis infection in red foxes.

The validity of a coproantigen ELISA for Echinococcus multilocularis was evaluated by comparison of three diagnostic methods; autopsy, egg examination and the ELISA. Of 71 foxes, 39 were found to be infected with the cestode at autopsy. The overall mean of worm burdens was 3,451, but the number varied (1-34,522). The ELISA could detect 94.9% (37/39) of the worm positives and there were no false-positives. Two false-negatives were infected with 1 and 4 cestodes, whereas 3 cases with similar worm burdens (2, 4 and 6 worms) were diagnosed as positives. This indicates the detection limit of the assay may be equivalent to less than 10 (in the worm burden). On the other hand, egg examination showed low sensitivity (43.6%, 17/39). These results suggest the ELISA has a potential to replace for the conventional methods.     

Use of ELISA to assess lungworm infection in calves

The enzyme-linked immunosorbent assay (ELISA) technique was applied to the detection of lungworm infections in calves. In experimentally infected animals different responses to larval and adult worm antigens were observed. The response to adult worm antigens was delayed in vaccinated animals when infection occurred by the gradual uptake of infective larvae from contaminated pasture. A serological survey in The Netherlands demonstrated a high incidence of lungworm infection in both vaccinated and unvaccinated herds. There was a good correlation between anti-adult worm and anti-larval ELISA-titres. ELISA appeared to be a useful technique for assessing the level of lungworm infection in a herd.     

Serologic prevalence of selected infectious diseases in cats with uveitis.

Serologic evidence of infection by Toxoplasma gondii, feline leukemia virus, feline coronaviruses, or feline immunodeficiency virus (FIV) is commonly found in cats with uveitis. Serum samples from 124 cats with uveitis were assayed by use of ELISA for the detection of T gondii-specific immunoglobulin M (IgM), IgG, and circulating antigens (Ag), as well as an ELISA for feline leukemia virus Ag, an ELISA for antibodies to FIV, and an indirect fluorescent antibody assay for antibodies to feline coronaviruses. Serologic evidence of infection by 1 or more of the infectious agents was detected in 83.1% of the samples. Serologic evidence of T gondii infection, defined as the detection of T gondii-specific IgM, IgG, or Ag in serum, was found in 74.2% of the samples. The seroprevalence of T gondii infection was significantly greater in cats with uveitis than in healthy cats from a similar geographic area. Serum samples from cats with serologic evidence of both T gondii and FIV infections were more likely to contain T gondii-specific IgM without IgG than samples from cats with serologic evidence of T gondii infection alone. Cats with serologic evidence of FIV and T gondii coinfection had a higher T gondii-specific IgM titer geometric mean and a lower T gondii-specific IgG titer geometric mean than did cats with serologic evidence of T gondii infection alone. Serologic evaluation for T gondii infection should include assays that detect IgM, IgG, and Ag, particularly in cats coinfected with FIV.     

犬细粒棘球绦虫粪抗原夹心ELISA检测方法的建立

为建立特异性和敏感性高的检验犬细粒棘球绦虫感染的方法。用细粒棘球绦虫(简称,Eg)成虫抗原分别免疫兔和绵羊,收集高免血清,纯化的高免抗体。依据抗体夹心ELISA工作原理,以兔抗体包被,检测感染Eg、不同犬带科绦虫的实验犬和空白犬粪样,绵羊抗体扑捉抗原,HRP标记兔抗绵羊IgG(1∶8 000)催化显色,用酶标仪测定OD 405nm吸光度,用以确定其特异性和敏感性。试验结果表明,敏感性为82.69%(43/52),特异性为85.88%(140/163);粪抗原在感染细粒棘球绦虫16d后可检出,最低抗原浓度为9.7ng/mL即犬感染5条成虫时可检测出阳性。该检测方法具有较好的特异性和灵敏性,为进一步研制检测细粒棘球绦虫虫体抗原ELISA检测试剂盒奠定了基础。     

The use of Ag85 complex as antigen in ELISA for the diagnosis of bovine tuberculosis in dairy cows in Brazil

Several enzyme-linked immunosorbent assay (ELISA) methods have been investigated to evaluate their performance in the diagnosis of tuberculosis in cattle. Increased production of antibodies to the proteins of antigen 85 complex (Ag85) after experimental and natural infection in cattle shows that they are strongly immunogenic in vivo. The purpose of this study was to evaluate the use of Ag85 as an antigen in ELISA for the diagnosis of bovine tuberculosis in dairy cows in Brazil. The test groups consisted of 46 serum samples from intradermal tuberculin test (ITT)-positive animals (Group A) and 46 samples from ITT-negative animals (Group B). Group C comprised 12 samples from a tuberculosis-free herd and was the control group of the study. Samples were tested in an ELISA using Ag85 as antigen. Differences between the mean ODs of groups A and B and A and C were significant (P < 0.01), but no significant difference (P > 0.05) was observed between groups B and C. The sensitivity of the ELISA using Ag85 was 91.3% (42/46) and its specificity was 94.8% (55/58). These results were not significantly different (P > 0.05) from those observed in a previous study of an ELISA using purified protein derivative (PPD). We concluded that, although Ag85 can be used as antigen for ELISA tests in the diagnosis of bovine tuberculosis with good sensitivity and specificity rates, no significant advantages were observed in relation to the ELISA using PPD that could justify the purification and utilization of Ag85 as a single antigen in routine methods of diagnosis.     

Development of a copro-antigen capture ELISA for detecting Ostertagia ostertagi infections in cattle

The present study reports on the development of a copro-antigen capture ELISA for detecting Ostertagia ostertagi infections in cattle. The ELISA was based on polyclonal rabbit antibodies, which recognize O. ostertagi excretory/secretory antigens (ES). ES antigens are released by the metabolic active stages of the parasite in the abomasum, and passed in the faeces of the host. The detection limit of pure ES material was 30 ng ml(-1) in sample buffer and 125 ng ml(-1) in faecal extract. The test was evaluated using a follow up from six artificially infected calves. Elevated levels of Ostertagia coproantigens could be measured from 21 days after infection, indicating that only the presence of adult parasites can be detected. To evaluate the capacity of the assay to measure levels of infection, three groups of cattle were tested: 38 artificially infected calves, 17 naturally infected first grazing season calves and 16 naturally infected adult dairy cows. Optical densities were significantly correlated to the worm burdens of the animals and the ELISA had an overall sensitivity of 91% and a specificity of 45%. The test gave negative readings for faeces of animals carrying patent mono-infections with Cooperia oncophora.     

Evaluation of recombinant fructose-1,6-bisphosphate aldolase ELISA test for the diagnosis of Schistosoma japonicum in water buffaloes

Fructose-1,6-bisphosphate aldolase (FBPA) is an ubiquitous enzyme essential for glycolysis, gluconeogenesis and the Calvin cycle. It has been demonstrated to induce immune responses and to be useful in the immunodiagnosis of malaria. In this study, FBPA was cloned from the adult worms of Schistosoma japonicum and tested as an antigen for the diagnosis of S. japonicum infection in water buffaloes. Enzyme-linked immunosorbent assay (ELISA) was performed on the sera from 32 infected water buffaloes and 20 negative controls using the recombinant FBPA protein or soluble worm antigen preparation (SWAP) as an antigen. The OD cut-off values were determined to be 0.57 with 100% specificity and 100% sensitivity for the FBPA ELISA and 1.13 with 93.8% specificity and 95.0% sensitivity for the SWAP ELISA. These findings indicate that the recombinant FBPA of S. japonicum should be an useful diagnostic tool for the detection of antibodies against S. japonicum.     

Evaluation of recombinant fructose-1,6-bisphosphate aldolase ELISA test for the diagnosis of Schistosoma japonicum in water buffaloes

Fructose-1,6-bisphosphate aldolase (FBPA) is an ubiquitous enzyme essential for glycolysis, gluconeogenesis and the Calvin cycle. It has been demonstrated to induce immune responses and to be useful in the immunodiagnosis of malaria. In this study, FBPA was cloned from the adult worms of Schistosoma japonicum and tested as an antigen for the diagnosis of S. japonicum infection in water buffaloes. Enzyme-linked immunosorbent assay (ELISA) was performed on the sera from 32 infected water buffaloes and 20 negative controls using the recombinant FBPA protein or soluble worm antigen preparation (SWAP) as an antigen. The OD cut-off values were determined to be 0.57 with 100% specificity and 100% sensitivity for the FBPA ELISA and 1.13 with 93.8% specificity and 95.0% sensitivity for the SWAP ELISA. These findings indicate that the recombinant FBPA of S. japonicum should be an useful diagnostic tool for the detection of antibodies against S. japonicum.     

牛乳中黄曲霉毒素B_1和M_1检测方法的比较

比较了免疫亲和柱-高效液相色谱法(HPLC)和酶联免疫吸附法(ELISA)测定牛乳中黄曲霉毒素B1(AFB1)和M1(AFM1)的回收率、方法检出限及精密度。结果表明,HPLC法测定AFB1、AFM1的检测限分别为0.02和0.01μg·L-1,ELISA法测定AFB1、AFM1的最低检出限分别为0.05和0.02μg·L-1;阴性牛乳试样AFB1(0.003、0.006、0.012、0.024μg·kg~(-1))和AFM1(0.005、0.01、0.025、0.05μg·kg~(-1))的回收率试验表明,HPLC法测定AFB1与AFM1的回收率分别为77.58%~82.81%和77.51%~82.23%,变异系数分别为1.93%和3.51%;ELISA法测定AFB1与AFM1回收率分别为77.28%~79.11%和75.40%~76.34%,变异系数分别为2.20%和3.80%,两种方法测定AFB1的回收率差异不显著(P0.05),当AFM1添加浓度大于0.025μg·kg~(-1)时,HPLC法回收率显著高于ELISA法(P0.05);综上,两种方法灵敏度高、重复性好,HPLC法测定高浓度AFM1时准确性优于ELISA法。     

Quantitation of canine plasma von Willebrand factor antigen using a commercial enzyme-linked immunosorbent assay.

The purpose of this study was to evaluate a commercial enzyme-linked immunosorbent assay (ELISA) for human von Willebrand factor antigen (vWF:Ag) with respect to its potential value in quantitating the protein in canine plasma. The assay was a sandwich technique using F(ab')2 fragments specific for von Willebrand factor (vWF) and a peroxidase conjugated rabbit anti-vWF second antibody, with a microplate as the support surface. Canine plasmas were assayed by ELISA, and by Laurell electroimmunoassay (EIA), our reference methodology. The ELISA had a within-day variation of 1.21-4.44% and a between-day variation of 0.85-4.88% depending on the level of vWF:Ag. The sensitivity of the assay was less than 0.1% vWF:Ag. The range of vWF:Ag concentrations in plasmas from 24 clinically normal dogs compared favorably with the range for the same plasmas when assayed by EIA (ELISA = 60-152% of normal; EIA = 50-142% of normal). In 121 canine plasmas with vWF:Ag concentrations (as assessed by EIA) ranging from undetectable levels (less than 6% of normal) to 142% of normal, there was good correlation with measurements made by ELISA (correlation coefficient = 0.835). It was concluded that this commercial ELISA technique could be used to provide reliable, same-day measurements of canine plasma vWF:Ag. Since it requires no special equipment other than a microplate reader and washer it is particularly suitable for laboratories lacking the electrophoretic expertise or equipment required for EIA.