GAg-ELISA检测丝虫病的方法与应用

ＧＡｇ－ＥＬＩＳＡ检测丝虫病的方法与应用徐礼周，冯子玲，叶萍，钟志萍，周勇志（中国农业科学院上海家畜寄生虫病研究所）ＥＬＩＳＡ技术具有灵敏度高、特异性强、设备简单、操作简便、试剂稳定、对人体无害等优点，是当今最先进的免疫诊断方法之一［“。目前国际上Ｅ...     

一种检测猪生殖——呼吸道综合征的ELISA抗原的制备

利用ＴｒｉｔｏｎＸ－１００助溶，由猪生殖－－呼吸道综合征病毒（ＰＲＲＳＶ）感染的ＭＡＲＣ－１４５细胞可制备出高纯度的ＥＬＩＳＡ抗原。这一技术消除了涉及到ＰＲＲＳＶ抗的及细胞抗在的常见本底反应。将这种高质量抗原应用到检测抗ＰＲＲＳＶ抗体的间接ＥＬＩＳＡ中，辅以一咱有效的血清封闭稀释剂可消除血清样品的非特异性反应。这种ＥＬＩＳＡ技术比间接免疫荧光试验（ＩＦＡ）更为敏感；特别是对感染后期血清有高度特异性     

快速检测鸡新城疫病毒的聚乙二醇单抗夹心ELISA试验的建立和应用

以抗鸡新城疫病毒（ＮＤＶ）单抗夹心ＥＬＩＳＡ试验为基础，在６０００，建立了ＰＥＧ－ＥＬＩＳＡ法，使用此法检测临诊样品，与单抗夹心ＥＬＩＳＡ相比，时间缩短７０分钟且提高了ＯＤ４９０值，易于识别。结果表明，ＰＥＧ－ＥＬＩＳＡＩ法具有实际应用价值。ＰＥＧ能加强抗原－抗体反应的速率和强度，这种效应在固相夹心ＥＬＩＳＡ第二步表现尤其明显。ＰＥＧ的这种非特异性效应对于检测其它抗原或抗体的ＥＬＩＳＡ试验可能具有     

现代诊断技术在兽医学上的应用及其进展

1　免疫酶技术 (ＥＩＡ)ＥＩＡ是根据抗原 -抗体结合的特异性 ,以酶作标记物 ,酶对底物具高效催化作用的原理而设计 ,既可用于抗原的检查 ,也常用于血清抗体的检测。1 .1　酶联免疫吸附试验 (ＥＬＩＳＡ)ＥＬＩＳＡ是应用最广泛的免疫酶技术 ,包括间接法、夹心法、竞争法等。自ＶＯＬＬＥＲ和ＢＩＤＷＥＬＬ(1 975 )首先报道了应用间接ＥＬＩＳＡ方法检测病毒特异性抗体以来 ,ＥＬＩＳＡ方法已普遍应用于传染病的流行病学普查、抗体水平的评价以及疫苗质量的监控和标化等 ,迄今为止大多数动物病原体均已发展或报道了ＥＬＩＳＡ方法的研究与…     

应用Dot—ELISA检测鸡白血病病毒群特异性抗体

利用斑点酶联免疫吸附试验（Ｄｏｔ－ＥＬＩＳＡ）的基本原理,根据蛋清可以直接吸附在硝酸纤维素（ＮＣ）膜上的特点,建立了Ｄｏｔ－ＥＬＩＳＡ检测鸡白血病病毒（ＡＬＶ）群特异性抗原的方法,并与双抗体夹心酶联免疫吸附试验（ＤＡＳ－ＥＬＩＳＡ）进行比较,两种方法具有相同的特异性和敏感性。     

银加强胶体金技术检测猪细小病毒的研究

本研究首次成功地建立了检测猪细小病毒（ＰＰＶ）抗原的银加强胶体金检测法（ＳＥＣＧＡ），并确定了操作流程中的最佳试验条件。应用该法对纯化ＰＰＶ抗原的最低检出量为０．３１２５ｎｇ／点，其敏感性分别为间接Ｄｏｔ－ＥＬＩＳＡ和ＨＡ的８倍和１０００倍。特异性阻断试验和交叉反应试验证明ＳＥＣＧＡ具有较高的特异性。２０份样本ＳＥＣＧＡ的检测结果与病毒分离和鉴定结果完全相符。ＳＥＣＧＡ和间接Ｄｏｔ－ＥＬＩＳＡ对７７份样本检测的阳性率分别为８３．１％和８０．５％，其阳性符合率为９６．９％。研究结果表明本法具有经济、敏感性、特异性强等优点，可用于ＰＰＶ感染的特异诊断。为胶体金技术应用于畜禽传染病的诊断和研究奠定基础。     

抗原捕捉ELISA与常规方法滴定IBDV的比较

作者建立了两种抗原捕捉酶联免疫吸附试验方法（多抗和单抗ＡＣ－ＥＬＩＳＡ），用于滴定用不同宿主系统增殖的传染性氏囊毒病，这些宿主系统包括ＢＧＭ－７０传代细胞系。鸡胚成纤维细胞和鸡法氏囊，两种检测方法都有较高的特异性，但是多克隆ＡＣ－ＥＬＩＳＡ比单克ＡＣ－ＥＬＩＳＡ更加敏感（Ｐ〈０．０５），结果还表明，滴定ＩＢＤＶ抗原的常规方法（细胞培养物和鸡胚）比多抗ＡＣ－ＥＬＩＳＡ更敏感。     

Ab—ELISA和Ag—ELISA诊断骆驼伊氏锥虫感染的比较

采用四种方法：即①单克隆抗体检抗原的酶联免疫吸附试验（Ａｇ－ＥＬＩＳＡ）②检测抗体的在酶联免疫吸附试验（Ａｂ－ＥＬＩＳＡ）③表膜检查（ＢＣＥ）④小白鼠接种（ＭＩ），检查了肯尼亚１８３头骆驼锥虫的循环抗原，ＢＣＥ查出尼亚１８３头骆驼锥早的循环抗原，ＢＣＥ查出３７头阳性２０％），ＭＩ６０头阳性（３３％），Ａｇ－ＥＬＩＳＡ６３头阳性（４９％），Ａｂ－ＥＬＩＳＡ检出出的２４头中，Ａｇ－ＥＬＩＳＡ检出１８头     

单抗—ELISA一步法检测兔粪样中的轮状病毒

用单抗２Ｂ４包被ＥＬＩＳＡ微孔反应板，将处理后的粪样和酶标兔抗ＬａＲＶＩｇＧ同时加入包被孔内孵育，置微型振荡器上振摇，建立了检测粪样中兔轮状病毒抗原的单抗－ＥＬＩＳＡ一步法快速诊断技术，与常规ＥＬＩＳＡ，病毒分离培养，电镜，荧光抗体等作了实验室和临床应用比较，结果证明单抗－ＥＬＩＳＡ－一步法第三特异，快速，简便１，适于基层临床推广应用。     

诊断B组轮状病毒的三种酶联免疫方法的建立

用从当地分离鉴定的山羊Ｂ组轮状病毒ＫＢ－６３毒株，在剥夺初乳的山羊羔体内传代繁殖病毒，制备抗原，建立了酶联免疫吸附试验（ＥＬＩＳＡ）、斑点酶联免疫吸附试验（Ｄｏｔ－ＥＬＩＳＡ）及生物素－亲和素系统酶联免疫吸附试验（ＢＡＢ－ＥＬＩＳＡ）。这三种方法经阻断试验、重复性试验以及交叉试验表明，其特异性、重复性好。以双盲法将此三种方法与对流免疫电泳法（ＣＩＥ）和聚丙烯酰胺凝胶电泳法（ＰＡＧＥ）作了对比研究，     

应用酶标SPA Dot-ELISA同时检测牛羊血吸虫病和肝片吸虫病

采用给雄性家兔注射黄牛、水牛、山羊ＩｇＧ混合液（１∶１∶１）制备的兔抗黄牛、水牛和山羊ＩｇＧ连接酶标ＳＰＡ，解决了酶标ＳＰＡ与牛羊血吸虫和肝片吸虫免疫复合物相结合的技术关键，并制成了可同时用于牛羊血吸虫病和肝片吸虫病诊断的酶试剂。建立了Ｄｏｔ－ＥＬＩＳＡ同时检测牛羊血吸虫病和肝片吸虫病技术，能测出人工感染１～１５０条虫体７～４２ｄ病畜抗体，阳性检出率达１００％。     

Serodiagnosis of Fasciola gigantica infection in buffaloes

The agar gel precipitation test (AGPT), counter immunoelectrophoresis (CIEP) and indirect haemagglutination (IHA) were evaluated for the diagnosis of fascioliasis due to Fasciola gigantica in buffaloes. The sensitivity of these tests varied with the intensity of infection; and was greatest when the fluke burden in liver exceeded 100. CIEP detected 76.06% of infected sera and was most sensitive, followed by IHA which detected 68.37% of the infected sera. The AGPT was found to be least sensitive, detecting only 57.4% of the infected sera. Although these tests were limited by the occurrence of false-positive reactions, their use may be an aid for effective diagnosis of fascioliasis in buffaloes.     

穴位接种IBD双价细胞苗鸡的抗原定位及免疫活性细胞监测

１５日龄滨白鸡后海穴接种鸡传染性法氏囊病（ＩＢＤ）双价细胞苗，采用免疫荧光组化跟踪抗原定位和免疫活性细胞的免疫形态学监测，以评价其免疫效果。同时与口服和肌肉免疫途径进行了对比试验。结果表明，后海穴穴位免疫优于口服免疫和肌肉免疫。本文对穴位免疫机制进行了探讨     

Studies on the pathology of dicrocoeliasis and fascioliasis in the goat. I. The histopathology of the liver and bile ducts

Comparative studies were made on the histopathology of the liver and bile ducts in dicrocoeliasis and fascioliasis of the goat. Investigations revealed certain similarities in the nature of the pathological phenomena produced by D. dendriticum and F. hepatica. However, a feature peculiar to fascioliasis was the migratory tracks of immature flukes in the liver parenchyma. Further, the lesions produced by fascioliasis were distinctly more pronounced than those present in dicrocoeliasis.In both parasitic diseases the bile-duct walls showed glandular hyperplasia, increased frequency of goblet cells and globule leucocytes, and extensive fibrosis in the outer layers. The desquamation produced by Dicrocoelium remained superficial whereas Fasciola was capable of causing deep erosions in the walls of the main bile ducts. The bile-duct walls did not become calcified. Increased fibrosis was present in the interlobular areas, too, but only fascioliasis produced extensive disorganisation of the hepatic architecture, due to the scars originating from repaired migratory tracks. The liver parenchymal cells were degenerated, showed decreased glycogen but were increasingly infiltrated by neutral fats.Keyword: Dicrocoelium dendriticum, Fasciola hepatica, liver, bile ducts     

A study of a restricted programme of strategic dosing against Fasciola hepatica with triclabendazole

A restricted programme of strategic dosing with triclabendazole was used to treat a sheep flock with severe clinical fascioliasis. After five years the percentage of ewes passing fluke eggs was reduced from 49 per cent to less than 1 per cent, without the reappearance of clinical fascioliasis. It is suggested that this programme offers an efficient and practical means of controlling fascioliasis.     

Comparative evaluation of the enzyme-linked immunosorbent assay (ELISA) in the diagnosis of natural Fasciola gigantica infection in cattle

The enzyme-linked immunosorbent assay (ELISA) was evaluated for the diagnosis of naturally acquired Fasciola gigantica infection in cattle in comparison with conventional parasitological techniques. Using unfractionated whole worm extract of F. gigantica as the antigen, it was observed that there was a good correlation between ELISA positivity and positive diagnosis of fascioliasis by post mortem liver examination, bile egg sedimentation and faecal egg sedimentation. There was, however, a disparity between ELISA results and faecal egg flotation results.     

Influence of 17beta-oestradiol, nortestosterone and dexamethasone on the adaptive immune response in veal calves

In veal calf production androgens, estrogens and glucocorticoids are used to stimulate growth. However, sexhormones and glucocorticoids also influence the function of the immune system. From studies in humans and mice, androgens are known as immunosuppressive, while estrogens stimulate the production of antibodies and glucocorticoids also enhance the T-helper 2 response. To investigate whether the adaptive immune system is influenced by hormone administration, calves were treated with a hormone cocktail containing androgens, estrogens and glucocorticoids and vaccinated against Mycobacterium avium spp. paratuberculosis. The activity of the adaptive immune system was measured by using an antigen specific elispot assay (ES), lymphocyte stimulation test (LST) and an enzyme-linked immuno sorbent assay (ELISA). The results showed that the hormone treatment did not lead to significant differences in the function of the adaptive immune system between the hormone treated and the not hormone treated group while growth was stimulated in the hormone treated group.     

Prevalence of bovine fascioliasis in a new-emerging focus of human fascioliasis in BoyerAhmad district,southwest of Iran

BackgroundThe prevalence of fascioliasis in a population of cattle in a new-emerging focus of human fascioliasis in Boyer-Ahmad District, in the southwest of Iran, was investigated, using an ELISA system and coprological methods.MethodsBlood, as well as stool samples, were collected from 150 dairy cattle consisted of 82 (54.7%) males and 68 (45.3%) females, aged from 1 to 8 years, selected from different pastures. The stool samples were evaluated by direct wet mount microscopic examination as well as sedimentation methods. Moreover, modified Telemann was used for detecting of Fasciola eggs. Anti-Fasciola antibodies in the cattle sera samples were determined, using a Fasciola excretory-secretory antigen-specific ELISA.ResultsAnti-Fasciola antibodies were detected in 64 (42.4%) of the subjects by ELISA. Out of 64 seropositive cases, 29 (45.3%) were female and 35 (54.6%) were male. No significant association was found between sex and seropositivity to fascioliasis. Considering the age of the cattle, the highest prevalence of fascioliasis (23%) was found in the 5-year old age group. However, the difference between the age of the cattle and seropositivity to fascioliasis was not statistically significant (P > 0.05). Fasciola eggs were not detected in fecal samples of any of the studied cattle, while Paramphistomide, as well as Dicrocoelium dendriticum eggs, were detected in 6 (4%) and 17 (11.3%) of the subjects, respectively.ConclusionFindings of the current study provide basic information about bovine fascioliasis in a new focus of human fascioliasis in Iran, which is needed for effective control of this parasitic infection in such areas.     

牛肝片形吸虫病的普查与防治

牛肝片形吸虫病是海口市东北部地区养牛最常见的一种寄生虫病。在海口市东北部地区的府城、灵山、龙塘、旧州等乡镇由于该地区河流小溪多,水塘水域多,终年积水的水田多,且肝片形吸虫的中间宿主锥实螺在该地区水域广泛分布和农民有在水田、水边放牧的习惯,所以在该地区感染牛肝片形吸虫病的牛很多。本文结合牛肝片形吸虫病的流行病学调查及防治等方面的情况,以及对当前海口市东北部地区流行的牛肝片形吸虫病的一些常见问题进行分析,提出了一些解决方法和对策,为该地牛肝片形吸虫病的有效防制提供参考。     

Antibody response of calves to a single infection of Fasciola gigantica determined by an indirect fluorescent antibody technique.

The antibody response of six calves infected with 500 metacercariae of Fasciola gigantica each was monitored throughout 30 weeks of infection using an indirect fluorescent antibody technique (IFA). In vitro excysted F gigantica were employed as the test antigen. All animals showed high antibody titres from two to six weeks post-infection. Thereafter the antibody titres diminished gradually. Although all the experimentally infected animals harboured fluke burdens at autopsy, most gave negative IFA tests from 22 weeks post infection onwards. Specific immunofluorescent staining occurred on the surface glycocalyx of the newly excysted flukes. It is likely that this glycocalyx provides one of the earliest antigenic stimuli for the host's immune reactions to fascioliasis.