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1.
用重组杆状病毒表达的钎生殖和呼吸综合征病毒（ＰＲＲＳＶ）Ｎ蛋白作为抗原，建立了诊断猪生殖和呼吸综合征（ＰＲＲＳ）的Ｉ－ＥＬＩＳＡ，与ＩＤＥＸＸ公司试剂盒检测结果的符合率为９７．３％，与间接 光抗体试验９ＩＦＡ）的符合率为１００％。用该方法的检测国内猪血清８５份，阳性率为１８．８％；能检出ＰＲＲＳ疫苗免疫猪６ｄ的血清抗体，；与猪瘟、伪狂犬病、猪巴氏杆菌病、猪霉形体病阳性血清无交叉反应。  相似文献   

2.
用蔗糖不连续梯度纯化的减蛋综合征＊（ＥＤＳ－７６）病毒包被ＥＬＩＳＡ板，建立了检测ＥＤＳ－７６抗体的间接ＥＬＩＳＡ法。经测定，抗原最适包被浓度为１μｇ／ｍＬ，待检血清最佳稀释度为１：２００，以ＯＤ４９０〉０．３，Ｐ／Ｎ〉２．１判为阳性结果。对４个鸡场的１２０份鸡血清进行检测，阳性率分别为０，４３．３３％，８０．００％和９３．３３％。  相似文献   

3.
酶联免疫吸附试验检测猪生殖与呼吸综合征抗体的研究   总被引：7，自引：1，他引：6  
建立的检测猪生殖与呼吸综合征（ＰＲＲＳ）抗体的酶联免疫吸附试验（ＥＬＩＳＡ），其抗原包被浓度为１．７ｍｇ／Ｌ，血清的最适稀释度为１：１００，酶标兔抗猪ＩｇＧ的家访工为１：２０００。比较试验表明，ＥＬＩＳＡ的敏感性优于间接免疫荧光试验（ＩＦＡ）。用ＥＬＩＳＡ检测１９９１年从美国进口猪的血清７６份，阳性率为零；１９９５年从加拿大进口猪的血清１６３份，阳性率为５．５２％；北京地区甲猪场的血清１２１份，阳  相似文献   

4.
在内蒙古牙克石市图里河镇非马传贫注苗区，用琼脂扩散试验（ＩＤ）和酶联免疫吸附试验（ＥＬＩＳＡ）两种方法对６４匹马进行了对比检查，检出ＩＤ、ＥＬＩＳＡ双阳性４匹、ＤＬＩＳＡ单阳性马３匹。对３匹ＩＤ阴性，ＥＬＩＳＡ阳性马血清用不同浓度琼扩抗原重复检测结果为阴性，而用不同批次的ＥＬＩＳＡ抗原包被板及酶标抗体检测结果均为陧阳性，用马传盆病毒抗原对３匹单阳性马血清中和后，再进行ＥＬＩＳＡ测定，ＯＤ值均降低５  相似文献   

5.
Dot—ELISA检测猪生殖和呼吸综合征抗体的研究   总被引：10，自引：3，他引：7  
用差速离心法提纯ＰＲＲＳ病毒，利用ＮＣ膜作为载体，在国内外首次成功建立了检测ＰＲＲＳ血清抗体的斑点酶联免疫吸附试验（Ｄｏｔ－ＥＬＩＳＡ）。抗原包被浓度为１００μｇ／ｍｌ，被检血清工作浓度为１：１０，酶标兔抗猪ＩｇＧ工作浓度为１：６００。对７２份北京地区猪血清分别用Ｄｏｔ－ＥＬＩＳＡ和ＩＦ检测，Ｄｏｔ－ＥＬＩＳＡ检测阳性率为３３．３％，ＩＦ检测阳履率为３０．６％，与ＩＦ符合率较高，对部分待检血清检测  相似文献   

6.
用差速离心法提纯ＰＲＲＳ病毒，利用ＮＣ膜作为载体，在国内外首次成功建立了检测ＰＲＲＳ血清抗体的斑点酶联免疫吸附试验（Ｄｏｔ－ＥＬＩＳＡ）。抗原包被浓度为１００μｇ／ｍｌ，被检血清工作浓度为１∶１０，酶标兔抗猪ＩｇＧ工作浓度为１∶６００。对７２份北京地区猪血清分别用Ｄｏｔ－ＥＬＩＳＡ和ＩＦ检测，Ｄｏｔ－ＥＬＩＳＡ检测阳性率为３３．３％，ＩＦ检测阳性率为３０．６％，与ＩＦ符合率较高，对部分待检血清检测结果表明：６１份１９９１年进口美国猪血清阳性率为０％，１８２份１９９５年进口加拿大猪血清阳性率为４．９４％。本法不需特殊仪器，适用于基层兽医部门和猪场对该病的血清学诊断和流行病学普查  相似文献   

7.
应用改良阻断ELISA检测禽网状内皮组织增殖病血清抗体   总被引：3，自引：0，他引：3  
应用禽网状内皮组织增殖病病毒纯化抗原和抗ＲＥＶ单克隆抗体建立了改良阻断ＥＬＩＳＡ用于鸡血清中ＲＥＶ抗体检测，并对北京地区鸡群中随机采样的３６份血清样本进行了检测，阳性率为５．６％。与间接ＥＬＳＩＡ的检测结果进行了统计学比较，两种方法的阳性率无显著差异。结果表明本试验所建立的改良阻断ＥＬＩＳＡ可以用于鸡群ＲＥＶ感染的血清学调查。  相似文献   

8.
蓝舌病ＶＰ７抗原包被板在－２０℃保存期为６个月,４℃保存１个月。抗原批内重复性试验ＣＶ＜１０％,批间重复性试验ＣＶ＜１０％。ＶＰ７－ＥＬＩＳＡ与ＡＧＩＤ对４２０份血清平行检测结果：ＶＰ７－ＥＬＩＳＡ较ＡＧＩＤ试验多检出了 １３份阳性样品, ＶＰ,－ＥＬＩＳＡ检出的阳性样品对ＡＧＩＤ阳性样品的覆盖率为９７．４％,对采自陕西等省１８１６份牛、羊血清样品进行检测,检出阳性样品数为２５７份阳性检出率为１４．２％。  相似文献   

9.
应用ＳｅｐｈａｄｃｘＧ－２００层析法纯化鸡减蛋综合症病毒，利用ＮＣ膜作为载体，成功建立了检测ＥＤＳ－７６血请抗体的斑点酶联免疫吸附试验（Ｄｏｔ－ＥＬＩＳＡ）。抗原包被浓度为２μｇ／ｍｌ，被检血清浓度为１：２０，酶标兔抗鸡１ｇＧ浓度为１：２００．底物溶液最适ｐＨ值为７．２。对Ａ－Ｆ６个养禽场随机抽检血清１６０份，分别用Ｄｏｔ－ＥＬＩＳＡ、ＨＩ和ＡＧＰ检测，Ｄｏｔ－ＥＬＩＳＡ检出阳性率为５１．９％，ＨＩ检出阳性率为４６．９％，ＡＧＰ检出阳性率为３１．９％。对１４０日龄鸡人工感染试验，测定抗体消长规律。本方法不需特殊仪器。适用于基层兽医部门和养鸡场对ＥＤＳ－７６的血清学诊断和流行病学调查。  相似文献   

10.
旋毛虫病McAb快速ELISA诊断盒的研制与应用   总被引：7，自引：0，他引：7  
本研究建立了ＭｃＡｂ快速诊断猪旋毛虫病的ＥＬＩＳＡ试剂盒，应用旋毛虫单克隆抗体系和层析纯化抗原（ＰＡＡ）与旋毛虫肌幼虫排泄－分泌抗（ＥＳ）作常规ＥＬＩＳＡ平行检测６１头人工感染旋毛虫病猪血样，阳性率均为１００％，检测健康猪血样１０８２头，阴性率也为１００％，检测疫区自然感染猪血样１２５３头，阳性率分别为１．４４％和１．１２％，随机抽采１７５头猪血样和肉样作旋毛虫病消化法，常规法和快速法对比试验，诊  相似文献   

11.
Complement-fixing antibody to Mycoplasma hyopneumoniae in the serums of pigs experimentally infected with enzootic pneumonia was demonstrated by comparing the haemolytic titre of guinea-pig complement titrated in the presence of heated test serum, M. hyopneumoniae antigen and unheated normal pig serum with the titre obtained when the antigen was omitted. The haemolytic titres against sensitised sheep erythrocytes were determined after a fixation period of 16 to 18 hours at 5°C. When serums, collected at intervals of 3 to 7 days, from 43 pigs exposed to pigs experimentally infected with enzootic pneumonia were tested, 4.6 or more complement units were first fixed 14 to 44 (mean 23.4) days after contact began. Serums collected subsequently fixed from 4.6 to more than 31 complement units. This positive reaction usually persisted until the pigs were killed 4 to 35 weeks after contact began. Thirty-three had gross enzootic pneumonia lesions and 9 had lung lesions detected microscopically. Serum antibody was not detected in 73 weaned pigs aged 7 weeks in a pneumonia-free herd but serums from 9 of 15 unweaned piglets aged 9 to 14 days in the same herd, fixed between 3 and 7 complement units.  相似文献   

12.
Liu Q  Zhou YQ  Zhou DN  Liu EY  Du K  Chen SG  Yao BA  Zhao JL 《Veterinary parasitology》2007,143(3-4):260-266
Babesiosis has recently been recognized as an emerging infectious disease of buffalo in China. In order to investigate the epidemiology and enzootic potential of this parasite in Hubei province, we sought to develop a semi-nested PCR to detect Babesia orientalis in buffalo and the potential tick vector-Rhipicephalus haemaphysaloides by amplifying a specific 257bp fragment of B. orientalis 18S rRNA gene. The practical limit of detection showed that it had high sensitivity and an approximate parasitemia of 0.00000012% was detected by the PCR system. The blood samples of 121 asymptomatic buffaloes collected from four babesia endemic counties and that of 71 asymptomatic buffaloes collected from three babesia free counties in Hubei province of China were examined for the presence of B. orientalis using both Wright-Giemsa stained blood smear and semi-nested PCR. Microscopic examination revealed that 5/121 animals were positive, whereas 24/121 animals were positive by the semi-nested PCR assay. Of 378 ticks (R. haemaphysaloides) collected from buffaloes and examined by the semi-nested PCR, 35 were positive. The results showed that the semi-nested PCR was a useful method to investigate the epidemiology of buffalo babesiosis (B. orientalis), which is widely distributed in Hubei province, China.  相似文献   

13.
A capture operation to ascertain health status in free-ranging buffaloes from six different areas in the Caprivi Strip in the northeast corner of Namibia was conducted in October 2009. Basic information on the ticks and tick-borne pathogens normally found in wildlife from this area are scarce. The objective of this study was to assess the host status of African buffaloes, Syncerus caffer, for ixodid ticks and two selected tick-borne pathogens in the Caprivi Strip, a key area bordering Angola, Zambia, Botswana, and Zimbabwe. Four different tick species have been identified among the 233 collected specimens, and, of 95 tested buffaloes, 54 (57%) were positive for Theileria parva, whereas only 3 (3%) showed evidence of being infected with Ehrlichia ruminantium.  相似文献   

14.
Following primary infection of the eye, oral cavity, and/or nasal cavity, bovine herpesvirus 1 (BHV-1) establishes latency in trigeminal ganglionic (TG) neurons. Virus reactivation and spread to other susceptible animals occur after natural or corticosteroid-induced stress. Infection of calves with BHV-1 leads to infiltration of lymphocytes in TG and expression of IFN-gamma (interferon-gamma), even in latently infected calves. During latency, virus antigen and nucleic acid positive non-neural cells were occasionally detected in TG suggesting there is a low level of spontaneous reactivation. Since we could not detect virus in ocular or nasal swabs, these rare cells do not support high levels of productive infection and virus release or they do not support virus production at all. Dexamethasone (DEX) was used to initiate reactivation in latently infected calves. Foci of mononuclear or satellite cells undergoing apoptosis were detected 6h after DEX treatment, as judged by the appearance of TUNEL+ cells (terminal deoxynucleotidyltransferase-mediated dUTP-biotin nick end labeling). BHV-1 antigen expression was initially detected in lymphocytes and other non-neural cells in latently infected calves following DEX treatment. At 24h after DEX treatment, viral antigen expression and nucleic acid were readily detected in neurons. Our data suggest that persistent lymphocyte infiltration and cytokine expression occur during latency because a low number of cells in TG express BHV-1 proteins. Induction of apoptosis and changes in cytokine expression following DEX treatment correlates with reactivation from latency. We hypothesize that inflammatory infiltration of lymphoid cells in TG plays a role in regulating latency.  相似文献   

15.
A total of 236 urinary bladders (94 cattle and 142 buffaloes) collected from Bareilly, Uttar Pradesh, India, were studied for spontaneous lesions. These adult animals belonged to Institute’s organized dairy farm and rural areas in the Rohilkhand region of Uttar Pradesh. Grossly, congestion, hemorrhages, and cystoliths in urinary bladders were diagnosed. Histopathologically, the major conditions diagnosed were acute cystitis, 44 (18.64%), including, congestion, hemorrhages, sub-acute cystitis; chronic cystitis, 74 (31.35%), including chronic cystitis un-complicated type, lymphocytic cystitis, plasmolymphocytic cystitis, follicular cystitis, hyperplasia, nodular/acinar hyperplasia, and cystolithiasis; and nothing unusual diagnosed, 118 (50.00%). Similar types of pathological conditions were diagnosed in both species of animals with exception of follicular cystitis and nodular/acinar hyperplasia which was diagnosed respectively only in buffaloes and cystoliths in cows. In addition, a good number of 17/25 (68%) urinary bladder samples tested were found positive for presence of bovine papillomavirus type-2 (BPV-2) by polymerase chain reaction. These included eight cases of acute cystitis, an equal number of cases of chronic cystitis, and one normal bladder. BPV-2 is known as potential source of enzootic bovine hematuria along with other co-factors in enzootic areas. Lesions of zoonotic significance, like tuberculosis, etc., were not diagnosed. None of the observed lesions represented conditions, which, by themselves, would warrant carcass condemnation in buffaloes.  相似文献   

16.
苏皖水牛肝片吸虫和前后盘吸虫感染情况的调查   总被引：2，自引：0，他引：2  
苏皖两省１０县（市）共调查３０２农户的３０７头水牛,粪便学检查肝片吸虫阳性４３头,阳性率１４．０％,其中安徽的阳性率为１１．６１％（１８／１５５）,江苏阳性率为１６．４８％（２５／１５２）;母牛阳性率为１１．８％,公牛为１１．４％;前后盘吸虫阳性１７０头,阳性率为５５．４％（１７０／３０７）,其中安徽的阳性率为５８．１％（９０／１５５）;江苏的阳性为５２．６％（８０／１５２）,母牛的阳性率为６０．１％（１３７／２２８）,公牛的阳必国４１．８％（３３／７９）,两种吸虫的阳性率均呈现随年龄增加而提高的趋势,经统计学分析,两种吸虫的感染与地形无显著差异（Ｐ〉０．０５）,肝片吸虫的感染与性别、年龄亦无显著差异（Ｐ〉０．０５）,但对前后盘吸虫来说,感染与性别、年龄的差异显著（Ｐ〈０．０５）。而各县间两种吸虫感染则有显著关  相似文献   

17.
We conducted a seroepidemiological survey for antibodies to bovine leukemia virus (BLV), by using agar gel immunodiffusion technique, in dairy cows, water buffaloes, and yellow cattle throughout Taiwan. The positive reactors were 8.4% (376/4,459) in 1985 and 5.8% (1,277/22,190) in 1986, in 15 prefectures and 7 cities. Relatively high infection rate appeared in the northern and southern areas of Taiwan. Positive reactors increased gradually with age. The incidence of positive antibodies was 2 to 3 times higher in pasture-style farms than in housed-style farms. Among the 6,313 imported cattle, 302 (4.8%) showed positive reaction. Between 1985 and 1987, 5 cattle showed enzootic bovine leukosis among 351 sero-positive reactors in four highly positive prefectures. Survey of 134 water buffaloes and yellow cattle showed no positive reactors. This survey demonstrated that BLV-infection has increased over the years and spread throughout Taiwan.  相似文献   

18.
Despite low per-animal productivity of ruminants in developing countries, Johne's disease has not been investigated in buffaloes, which are primarily found in these countries. This is due to lack of expertise, diagnostic kits and priority to production diseases like Johne's disease. Presence of pathogenic Mycobacterium avium subspecies paratuberculosis (Map) was investigated by screening of target tissues (mesenteric lymph nodes and large intestine) by culture and IS 900 PCR, in 50 sacrificed buffaloes. Indigenous ELISA kit originally developed for goats and sheep was standardized in buffaloes and used to estimate sero-presence of Map in 167 serum samples representing population of buffaloes in Agra region of North India. In culture, 48.0% buffaloes were positive from 50 tissues each from mesenteric lymph nodes (34.0%) and large intestine (36.0%). IS 900 PCR was standardized using specific primers (150 C and 921) and 229 bp-amplified product was characteristic for Map. Of the 25 mesenteric lymph nodes, 40.0% were positive in IS 900 PCR. Genomic DNA from Map cultures was successfully amplified from all the 24 isolates (100.0%). Map was further genotyped as 'Bison type' using IS 1311 PCR-REA. Culture of tissues showed high presence of Map in target tissues, despite high culling rate in buffalos in view of high demand of buffalo meat. Specific tissue-PCR provided rapid confirmation of Map infection in sacrificed buffaloes. In tissue-PCR, all the cultures were positive as compared to 40.0% detected directly from tissues. ELISA kit using indigenous protoplasmic antigen was highly sensitive as compared to commercial antigen in detecting Map infection therefore, could be used as 'Herd Screening Test' in buffaloes against Johne's disease. This pilot study first time reports a highly pathogenic 'Bison-type' genotype of M. avium subspecies paratuberculosis from the riverine buffaloes (Bubalus bubalis) of Agra region in North India.  相似文献   

19.
Only limited information is available on the epidemiology and pathogenesis of Bovine Herpesvirus 1 (BoHV-1) in domestic buffalos. In this study, a virulent BoHV-1 field strain isolated from cattle was inoculated into buffaloes to evaluate their susceptibility to the virus and to investigate the establishment of viral latency through clinical, virological and serological investigations. Latency was also studied by attempting viral reactivation using pharmacological induction. Six of seven male, 5 months old buffaloes were intranasally inoculated with BoHV-1; the other animal was kept as negative control. The animals were clinically monitored during the post-infection (P.I.) and the post-pharmacological induction (P.P.) periods. During these periods, nasal and rectal swabs, and blood samples, with and without anticoagulant, were collected at 2–3 day intervals. On culling the animals, 206 days P.I., their trigeminal ganglia and tonsils were collected. No clinical signs referable to BoHV-1 were observed throughout the experimental period. However, seropositivity was detected in all infected animals within day 20 P.I., using BoHV-1 glycoprotein E and glycoprotein B competitive ELISAs (IDEXX) and virus neutralisation test. In real-time PCR (RT-PCR), five of these animals were positive, at least once, for nasal or rectal swabs, during the P.I. period. The sixth infected animal was found positive only in the trigeminal ganglia after culling. Ganglia were also positive for two other animals. Virus isolation in permissive cell-lines was successful for a part of the RT-PCR positive samples. The detected viruses were confirmed by genetic analysis as identical to the inoculated strain. No evidence of infection was observed in the negative control. This study represents the first experimental transmission of BoHV-1 in buffaloes, confirming their susceptibility to infection and their possible role as host/reservoirs of BoHV-1.  相似文献   

20.
A commercially available ELISA for detecting antibodies to liver fluke was evaluated for use in Australia. Milk and serum samples from cattle and sheep in which infection with Fasciola hepatica was confirmed by detection of eggs in faeces were used to estimate sensitivity. Similar samples collected from cattle and sheep outside the F. hepatica-endemic area were used to estimate specificity. The ELISA was also evaluated for detecting antibodies to F. hepatica in milk from sheep and antibodies to Fasciola gigantica in sera from cattle and buffaloes, but with small numbers of samples. In cattle, the sensitivity and specificity of the ELISA were 98.2% and 98.3% using serum and 97.7% and 99.3% using milk. In infected herds, 41.4% and 41.5% of animals were positive in the serum and milk ELISAs, respectively, whereas F. hepatica eggs were found in faecal samples from 26.5% of animals. In sheep, the sensitivity of the ELISA was 96.9% and the specificity was 99.4%. In infected flocks, 60.2% of animals were positive in the serum ELISA and F. hepatica eggs were found in faecal samples 52.2% of animals. There was perfect agreement in the ELISA between paired serum and milk samples collected from ewes. The assay detected antibodies in sera from cattle and buffaloes with natural and experimental F. gigantica infections. In the experimentally infected animals, antibodies were detected 2 weeks post-infection. We conclude that the ELISA will be a valuable tool for diagnosing F. hepatica infections in cattle and sheep. The assay may also be useful for diagnosing F. gigantica infections but further studies are required to establish sensitivity and specificity.  相似文献   

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