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1.
《畜牧与兽医》2017,(1):65-70
为检测弓形虫循环抗原,以实现弓形虫急性感染的早期诊断,本研究建立了基于ABC(avidin biotin-peroxidase complex)放大系统的双抗体夹心ELISA方法。通过制备弓形虫排泄分泌抗原(ESA)并免疫动物,经3种抗原的筛选以获得抗循环抗原(CAg)的单抗,以方阵滴定法确定最佳多抗包被浓度和单抗工作浓度,利用夹心法和ABC放大系统建立检测弓形虫循环抗原的夹心ABC-ELISA方法,用该方法对人工感染犬血清和其他阳性血清样本进行检测以确定该方法的检出时间和准确性,并应用于临床样品的检测。结果:获得了3株针对不同抗原表位的单克隆抗体,并对CAg有较高特异性。双抗体夹心ABC-ELISA反应条件:兔多抗包被浓度为3.7μg/mL,生物素标记3A5、3E5和10F5-3单抗在混合工作液中的浓度分别为0.1、0.13和0.12μg/mL。该ELISA方法对ESA最低检测限为11.9 ng/mL,并与隐孢子虫早期感染牛血清、血吸虫尾蚴早期感染牛血清、艾美耳球虫早期感染鸡血清、犬瘟热急性感染早期血清、犬细小病毒急性感染血清无交叉反应。用该ELISA方法检测人工感染的犬,于感染后2 d血清即显示阳性,该方法能明显区分标准阳性血清和阴性血清。用该方法检测68份猪临床血样(其中包括2份标准阳性血清),检测结果与Nest-PCR结果一致。表明该双抗体夹心ABC-ELISA方法特异性强、敏感性高,可用于弓形虫急性感染的早期或活动期的诊断。  相似文献   

2.
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。  相似文献   

3.
建立多房棘球绦虫粪抗原双抗体夹心ELISA检测方法,为犬感染多房棘球绦虫的早期诊断提供技术支撑。以5E10H5杂交瘤细胞株腹腔接种Balb/c鼠制备的腹水作为包被抗体,多房棘球绦虫成虫可溶性抗原免疫新西兰大白兔制备的多克隆抗体血清作为检测抗体,HRP标记的驴抗兔IgG作为二抗,建立双抗体夹心ELISA方法,检测试验犬(感染多房棘球绦虫)和阴性对照犬犬粪。结果显示,多房棘球绦虫成虫可溶性抗原具有良好的抗原性并能产生高效价抗体;对该方法的灵敏度进行检测,阳性粪样稀释至1∶10 000时仍显示为阳性;在感染动态分析中,该方法最早可在犬感染72 h后,最迟6 d后检测到多房棘球绦虫粪抗原。表明建立的方法可用于诊断犬多房棘球绦虫感染,为后期研制犬多房棘球绦虫粪抗原双抗体夹心ELISA检测试剂盒奠定了基础。  相似文献   

4.
为制备犬细小病毒特异性抗血清,用犬细小病毒DD株免疫健康易感比格犬,无菌采血后分离血清制备犬抗CPV-2血清。通过SN、ELISA、IFA、HI方法对制备的血清进行检测,未检出犬类常见的5种病毒抗体,说明该血清特异性良好。经测定,该血清中和抗体效价为1∶512;HI效价为1∶1280。应用结果表明,该血清对不同犬细小病毒毒株中和能力相当,对CPV-2感染犬有良好的治疗效果。  相似文献   

5.
为探究细粒棘球绦虫(E.granulosus,Eg)成熟虫体(MAW)特异性蛋白(EgM)(EgM9、EgM123)的抗Eg感染保护效果,本研究利用Eg疫苗候选抗原EgM9和EgM123重组蛋白,分别3次免疫实验犬,利用ELISA法检测免疫犬血清中IgG、IFN-γ和IL-10的变化情况,采用免疫组化法检测免疫犬肠系膜淋巴结和小肠中的特异性IgG抗体。结果显示,在第3次免疫后一周即免疫组犬抗体水平均最高时对其进行原头蚴攻击,攻毒后免疫组的抗体水平随之下降,且持续至第3次免疫后19周,免疫组均与对照组呈显著性差异(p0.05);第3次免疫后一周免疫组犬血清中IFN-γ和IL-10水平均呈升高趋势,且至第3次免疫后23周剖检,免疫组犬的肠系膜淋巴结和小肠组织经免疫组化法均检测出特异性IgG抗体。表明用EgM重组蛋白免疫犬,可诱导其产生IgG抗体而发挥抗Eg感染保护效果,为终末宿主犬包虫病的免疫机理探究和疫苗及相关检测试剂的研制奠定了基础。  相似文献   

6.
《中国兽医学报》2015,(11):1792-1798
RT-PCR扩增华支睾吸虫(Clonorchis sinensis)的Enolase基因后,进行原核表达,采用SDS-PAGE和Western blot检测表达产物;以纯化后的蛋白为包被抗原建立ELISA方法,并对工作条件进行优化。结果表明,ELISA最佳工作条件为:抗原最佳包被质量浓度为5mg/L,37℃1h再4℃过夜;5%脱脂奶粉,37℃封闭1h;待检血清1∶100稀释37℃孵育1h;酶标二抗1∶5 000稀释,37℃孵育45min;TMB显色作用时间10min;确定的阴阳性血清临界值为0.253。所建立的ELISA方法特异性较好,可检测犬华支睾吸虫病阳性血清,与犬卫氏并殖吸虫病阳性血清、犬蛔虫病阳性血清、犬弓形虫病阳性血清、犬新孢子虫病阳性血清均不发生反应。该方法敏感性为1∶3 200。批间批内重复性试验变异系数均小于10%。在临床应用中,对浙江地区353多份犬血清的检测表明,样品阳性率为1.13%。本研究建立的间接ELISA方法可以用于临床病例的血清学快速检测,为华支睾吸虫的血清流行病学调查提供了有效手段。  相似文献   

7.
寄生虫病是严重危害人畜健康的疾病,因其种类多,发病表现多样,病原诊断比较困难,检测感染宿主体内产生的抗体虽然对诊断有很好的辅助作用,但因其在治愈后仍能长期存在,容易造成误断。近年来已有不少关于检测感染宿主血液中循环抗原（circulatiflgantig6n,CAg）的报道,寄生虫循环抗原即活虫体排泄到宿主体内的大分子颗粒,主要是排泄分泌物或脱落物,具有抗原性,存在于各种组织内,含量与感染虫荷数相一致,CAg早于循环抗体（circulatingantithey,CAb）出现于宿主体内,因此检测血液中的CAg,不但能作早期诊断,而且能说明活虫…  相似文献   

8.
自然感染日本血吸虫病耕牛血清循环抗原消长规律的研究   总被引：1，自引：0，他引：1  
为了观察疫区自然感染日本血吸虫病耕牛血清循环抗原消长规律，本试验将血吸虫病非疫区的水、黄牛各１０头，转运至安徽省血吸虫病疫区，接受为期８周的自然感染。用斑点酶联免疫吸附试验测定牛血清循环抗原。结果，水、黄牛血清循环抗原滴度均于自然感染第４周后明显上升，水、黄牛自然感染第８周的血清循环抗原最高滴度分为１∶１０２４０和１∶５１２０，最低滴度则分别为１∶１２８０和１∶６４０，水牛循环抗原滴度高于黄牛。虫荷数与血清循环抗原滴度未见有相关性。  相似文献   

9.
为了观察绵羊用多头蚴抗原免疫及感染后的抗体消长规律,为羊脑多头蚴病的免疫预防和免疫诊断提供依据,本试验应用多头蚴原头节可溶性抗原、囊壁可溶性抗原、囊液粗抗原致敏绵羊红细胞对绵羊免疫3次及虫卵攻击感染后的血清抗体进行间接血凝试验（IHA）检测。结果表明,原头节抗原免疫组、囊壁抗原免疫组、囊液抗原免疫组及原头节ES抗原免疫组在首次免疫后1周,抗体滴度迅速升高,第3次免疫后1周达到峰值,虫卵感染后开始下降,到感染后30周接近正常水平。多头蚴3种抗原对同种抗原免疫组血清检测敏感性、特异性优于其它抗原,原头节免疫组、囊壁免疫组、囊液免疫组抗体水平明显高于原头节ES抗原免疫组。  相似文献   

10.
对试验猪用猪囊尾蚴细胞抗原免疫前、感染前、免疫后及用猪带绦虫卵感染后每隔10d经前腔静脉采血1次，分离血清，同时制备全血干血纸，分别用血清ELISA和干血纸ELISA进行检测。结果免疫组猪均在免疫后12d检出抗体；攻击感染组猪在攻击虫卵20d检出抗体，21d抗体水平达高峰，并一直持续到32周；阴性对照组的OD值一直在0.24～0.65。2种样品的检测结果基本一致。  相似文献   

11.
试验选用96头平均体重14.82 kg左右的杜×长×大断奶仔猪,随机分成4组,每组3栏,每栏8头(公母各半)。对照组饲喂基础日粮,试验1、2、3组分别添加1%80目白术、0.2%白术多糖和1%微米白术。试验期30 d。结果表明:在生长性能方面,与对照组相比,1%微米白术添加组可显著提高日增重(P0.05)、降低饲料增重比和腹泻率,而且效果优于1%80目白术组和0.2%白术多糖组,在肠道形态和肠道微生态区系方面,与对照组相比,日粮添加1%80目白术、0.2%白术多糖、1%微米白术均可不同程度的提高十二指肠和空肠的绒毛高度,加深十二指肠和空肠的隐窝深度,并且增加肠道微生态区系的多样性,其中以1%微米白术添加组的效果最佳。  相似文献   

12.
Sissay, M.M., Uggla, A. and Waller, P.J., XXXX. Prevalence and seasonal incidence of nematode parasites and fluke infections of sheep and goats in eastern Ethiopia. Tropical Animal Health and Production, XXXX. A 2-year abattoir survey was carried out to determine the prevalence, abundance and seasonal incidence of gastro-intestinal (GI) nematodes and trematodes (flukes) of sheep and goats in the semi-arid zone of eastern Ethiopia. During May 2003 to April 2005, viscera including liver, lungs and GI tracts were collected from 655 sheep and 632 goats slaughtered at 4 abattoirs located in the towns of Haramaya, Harar, Dire Dawa and Jijiga in eastern Ethiopia. All animals were raised in the farming areas located within the community boundaries for each town. Collected materials were transported within 24 h to the parasitology laboratory of Haramaya University for immediate processing. Thirteen species belonging to 9 genera of GI nematodes (Haemonchus contortus, Trichostrongylus axei, T. colubriformis, T. vitrinus, Nematodirus filicollis, N. spathiger, Oesophagostomum columbianum, O. venulosum, Strongyloides papillosus, Bunostomum trigonocephalum, Trichuris ovis, Cooperia curticei and Chabertia ovina), and 4 species belonging to 3 genera of trematodes (Fasciola hepatica, F. gigantica, Paramphistomum {Calicohoron} microbothrium and Dicrocoelium dendriticum) were recorded in both sheep and goats. All animals in this investigation were infected with multiple species to varying degrees. The mean burdens of adult nematodes were generally moderate in both sheep and goats and showed patterns of seasonal abundance that corresponded with the bi-modal annual rainfall pattern, with highest burdens around the middle of the rainy season. In both sheep and goats there were significant differences in the mean worm burdens and abundance of the different nematode species between the four geographic locations, with worm burdens in the Haramaya and Harar areas greater than those observed in the Dire Dawa and Jijiga locations. Similar seasonal variations were also observed in the prevalence of flukes. But there were no significant differences in the prevalence of each fluke species between the four locations. Overall, the results showed that Haemonchus, Trichostrongylus, Nematodirus, Oesophagostomum, Fasciola and Paramphistomum species were the most abundant helminth parasites of sheep and goats in eastern Ethiopia.  相似文献   

13.
1-(2-Chloroethyl)3-cyclohexyl-1-nitrosourea (CCNU) is an alkylating agent in the nitrosourea subclass. A prospective evaluation of CCNU was done to determine the maximally tolerated dosage of CCNU in tumor-bearing cats. Response data were obtained when available. Twenty-five cats were treated with CCNU at a dosage of 50-60 mg/m3 body surface area. Complete hematologic data were available for 13 cats. Neutropenia was the acute dose-limiting toxicity. The median neutrophil count at the nadir was 1,000 cells/microL (mean, 2,433 cells/microL; range, 0-9,694 cells/microL). The time of neutrophil nadir was variable, occurring 7-28 days after treatment, and counts sometimes did not return to normal for up to 14 days after the nadir. Based on these findings, a 6-week dosing interval and weekly hematologic monitoring after the 1st treatment with CCNU are recommended. The nadir of the platelet count may occur 14-21 days after treatment. The median platelet count at the nadir was 43,500 cells/microL. No gastrointestinal, renal, or hepatic toxicities were observed after a single CCNU treatment, and additional studies to evaluate the potential for cumulative toxicity should be performed. Five cats with lymphoma and 1 cat with mast cell tumor had measurable responses to CCNU. Phase II studies to evaluate antitumor activity should be completed with a dosing regimen of 50-60 mg/m3 every 6 weeks.  相似文献   

14.
Human and equine athletes are reported to have a high prevalence of gastric disease, and anecdotal evidence suggests a similar phenomenon applies to racing sled dogs. To investigate the prevalence of gastric disease in racing sled dogs, we conducted 2 gastroscopy studies on dogs competing in the annual Iditarod Sled Dog Race. A pilot study of dogs that were either dropped from the 2000 Iditarod Sled Dog Race because of illness or that finished the race indicated that, approximately 5 days after competing, 10 of 28 dogs (35%) had endoscopic evidence of gastric ulceration, erosion, or hemorrhage. The next year, an endoscopic study of 73 dogs participating in the 2001 Iditarod race was performed in order to evaluate a larger population of dogs. Data from 70 of these dogs could be used; 34 (48.5%) had ulceration, erosion, gastric hemorrhage, or some combination of these findings. When this group of 70 dogs was compared retrospectively to a control group of 87 dogs presented to the Texas A&M University (TAMU) Veterinary Medical Teaching Hospital, the Iditarod sled dogs had a significantly higher prevalence (P = .049) of gastric lesions. These findings suggest that, similar to athletes of other species, elite canine athletes have an increased prevalence of gastric disease compared to the canine population at large.  相似文献   

15.
Continuous rate infusion (CRI) of furosemide in humans is considered superior to intermittent administration (IA). This study examined whether furosemide CRI, compared with IA, would increase diuretic efficacy with decreased fluid and electrolyte fluctuations and activation of the renin-angiotensin-aldosterone system (RAAS) in the horse. Five mares were used in a crossover-design study. During a 24-hour period, each horse received a total of 3 mg/kg furosemide by either CRI (0.12 mg/kg/h preceded by a loading dose of 0.12 mg/kg IV) or IA (1 mg/kg IV q8h). There was not a statistically significant difference in urine volume over 24 hours between methods; however, urine volume was significantly greater after CRI compared with IA during the first 8 hours ([median 25th percentile, 75th percentile]: 9.6 L [8.9, 14.4] for CRI versus 5.9 L [5.3, 6.0] for IA). CRI produced a more uniform urine flow, decreased fluctuations in plasma volume, and suppressed renal concentrating ability throughout the infusion period. Potassium, Ca, and Cl excretion was greater during CRI than IA (1,133 mmol [1.110, 1,229] versus 764 mmol [709, 904], 102.7 mmol [96.0, 117.2] versus 73.3 mmol [65.0, 73.5], and 1,776 mmol [1,657, 2.378] versus 1,596 mmol [1,457, 1,767], respectively). Elimination half-lives of furosemide were 1.35 and 0.47 hours for CRI and IA, respectively. The area under the excretion rate curve was 1,285.7 and 184.2 mL x mg/mL for CRI and IA, respectively. Furosemide CRI (0.12 mg/kg/h) for 8 hours, preceded by a loading dose (0.12 mg/kg), is recommended when profound diuresis is needed acutely in horses.  相似文献   

16.
Genetic variations in chromosome Y are enabling researchers to identify paternal lineages, which are informative for introgressions and migrations. In this study, the male‐specific region markers, sex‐determining region‐Y (SRY), amelogenin (AMELY) and zinc finger (ZFY) were analysed in seven Turkish native goat breeds, Angora, Kilis, Hair, Honaml?, Norduz, Gürcü and Abaza. A SNP in the ZFY gene defined a new haplotype Y2C. All domestic haplogroups originate from Capra aegagrus, while the finding of Y1A, Y1B, Y2A and Y2C in 32, 4, 126 and 2 Turkish domestic goats, respectively, appears to indicate a predomestic origin of the major haplotypes. The occurrence of four haplotypes in the Hair goat and, in contrast, a frequency of 96% of Y1A in the Kilis breed illustrate that Y‐chromosomal variants have a more breed‐dependent distribution than mitochondrial or autosomal DNA. This probably reflects male founder effects, but a role in adaptation cannot be excluded.  相似文献   

17.
The purpose of this study was to determine the frequency of different tumor types within a large cohort of cats with intracranial neoplasia and to attempt to correlate signalment, tumor size and location, and survival time for each tumor. Medical records of 160 cats with confirmed intracranial neoplasia evaluated between 1985 and 2001 were reviewed. Parameters evaluated included age, sex, breed, FeLV/FIV status, clinical signs, duration of signs, number of tumors, tumor location(s), imaging results, treatment, survival times, and histopathologic diagnosis. Most of the cats were older (11.3 +/- 3.8 years). Primary tumors accounted for 70.6% of cases. Metastasis and direct extension of secondary tumors accounted for only 5.6 and 3.8% of cases, respectively. Twelve cats (7.5%) had 2 or more discrete tumors of the same type, whereas 16 cats (10.0%) had 2 different types of intracranial tumors. The most common tumor types were meningioma (n = 93, 58.1%), lymphoma (n = 23, 14.4%), pituitary tumors (n = 14, 8.8%), and gliomas (n = 12, 7.5%). The most common neurological signs were altered consciousness (n = 42, 26.2%), circling (n = 36, 22.5%), and seizures (n = 36, 22.5%). Cats without specific neurological signs were common (n = 34, 21.2%). The tumor was considered an incidental finding in 30 (18.8%) cats. In addition to expected relationships (eg, meninges and meningioma, pituitary and pituitary tumors), we found that lesion location was predictive of tumor type with diffuse cerebral or brainstem involvement predictive of lymphoma and third ventricle involvement predictive of meningioma.  相似文献   

18.
The epidemiology of H. placei and of other gastrointestinal nematodes in yearling dairy cattle was examined on two farms in Kiambu District, central Kenya during each of 13 one-month periods from April 1993 to April 1994. On each farm, 32 newly weaned dairy calves were given a single dose of albendazole and then placed on experimental pastures. Twelve of the animals were designated for bi-monthly slaughter (n = 2) and analysis of worm population characteristics and 20 were designated for blood and faecal collection and for weighing. Two parasite-free tracer calves were grazed alongside the weaner calves each month throughout the study period and were also slaughtered for analysis of worm populations. Faecal egg counts, haematological and serum pepsinogen determinations, herbage larval counts, and animal live weight changes were recorded monthly. The study revealed that Haemonchus placei, Trichostrongylus axei, Cooperia spp. and Oesophagostomum radiatum were responsible for parasitic gastroenteritis and that H. placei was the predominant nematode present in the young cattle on both farms. Faecal egg counts from resident cattle and necropsy worm counts revealed that pasture larval levels were directly related to the amount of rainfall. The total worm burdens in the animals were highest during the rainy season (March–June and October–December) and lowest during the dry seasons (July–September and January–February). The very low recovery of immature larvae of H. placei from the tracer calves indicated that arrested development is not a feature of the life cycle of this parasite in central Kenya. The maintenance of the parasite population depended on continuous cycling of infection between the host and the pasture. The agroclimatic conditions of the study area were such that, in general, favourable weather conditions for the development and survival of the free-living stages of gastrointestinal nematodes existed all year round.  相似文献   

19.
The occurrence of ectoparasites in sheep flocks is frequently reported but seldom quantified. Sheep production used to be a predominantly family activity in the state of Sa~o Paulo (Brazil), but it began to become a commercial activity in the past decade. Thus, information about the ectoparasites existing in sheep flocks has become necessary. The present data were obtained by means of questionnaires sent to all sheep breeders belonging to the `Associaça~o Paulista de Criadores de Ovinos' (ASPACO; Sa~o Paulo State Association of Sheep Breeders). Response reliability was tested by means of random visits paid to 10.6% of the respondents. Most of the properties (89.5%) reported the presence of one or more ectoparasites. Screw-worm (Cochliomyia hominivorax) was the most frequent ectoparasite (72.5%), followed by bot fly larvae (Dermatobia hominis, 45.0%), ticks (Amblyomma cajennense) and Boophilus microplus, 31.3%) and finally lice (Damalinia ovis, 13.8%). Combined infestations also occurred, the most common one being screw-worm with bot fly larvae (36.0%) followed by bot fly larvae with ticks (13.9%), screw-worm with ticks (9.3%), bot fly larvae with lice (6.9%), and ticks with lice (5.0%). The most common triple combination was screw-worm, bot fly larvae and ticks (12.8%). Breeds raised for meat or wool were attacked by bot fly larvae and ticks more often than other breeds. Lice were only absent from animals of indigenous breeds. The relationships among these ectoparasites are discussed in terms of sheep breeds, flock size, seasonality and the ectoparasitic combinations on the host.  相似文献   

20.
Comparison of different tests to diagnose feline infectious peritonitis   总被引：1，自引：0，他引：1  
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