共查询到20条相似文献，搜索用时 390 毫秒
1.
应用PCR检测成年鸭体内鸭瘟强毒的分布 总被引：8，自引：0，他引：8
鸭瘟病毒(DPV)强毒经人工接种和同居感染100日龄鸭后，应用聚合酶链反应(PCR)检测病毒在鸭体内各组织器官的动态分布。试验结果表明，DPV强毒经肌肉注射进入鸭体后6h可在肝、脾、血液和粪便中检测到DPV DNA；DPV强毒经肌肉注射到鸭体后各受检样品被检测到DPV DNA的先后顺序为：肝、脾、血液和直肠粪便(6h)→肺、脑和腿肌(12h)→肾和胸肌(24h)；同居鸭于混群后48h在肝、肺、血液和直肠粪便中检出DPV DNA；DPV强毒经同居感染鸭后各受检样品检测到DPV DNA的先后顺序为：肝、肺、血液和直肠粪便(48h)→脾和脑(72h)→胸肌和腿肌(96h)→肾(120h)。 相似文献
2.
用地高辛标记鸭圆环病毒（DuCV）全基因组克隆制成核酸探针,与鸭病毒性肝炎Ⅰ型病毒（DHV-Ⅰ）的cDNA、鸭瘟病毒（DPV）的DNA和DuCV的DNA进行斑点杂交以检测核酸探针的特异性,结果该探针只与DuCV的DNA杂交呈阳性。敏感性结果表明,该探针对DuCV的最低检出量约为5PgDuCV的DNA。应用该探针对从山东、江苏、四川等地采集的196只鸭病料,共980份脏器的组织DNA进行检测,结果个体阳性率为33．2％（65／196）,在各脏器中法氏囊和肝脏中DuCV检出率较高,检出率分别为52．3％和49．2％。同时对上述样品进行PCR检测,核酸探针检测与PCR检测2种方法的符合率为87．5％。结果表明,制备的DIG标记的核酸探针特异性强、敏感性高,适合对DuCV进行诊断和流行病学调查。 相似文献
3.
SYBR Green Ⅰ实时定量PCR快速检测鸭瘟病毒的研究 总被引：6，自引：0，他引：6
根据GenBank中鸭瘟病毒(DPV)UL6和UL7基因的保守序列,设计了一对特异性引物,扩增位于UL6和UL7基因第891～991位长度为101bp片断。以DPV标准强毒株DNA为模板,建立了SYBRGreenⅠ实时荧光定量PCR检测鸭瘟病毒的方法,用该方法检测鸭瘟标准强毒、疫苗毒及野毒株均为阳性,检测其他受试的非鸭瘟病毒DNA均为阴性,对鸭瘟强毒和疫苗毒人工感染鸭胚尿囊液的检出率均为100%,对6个临床送检样本的检出率为6/6,与病毒分离鉴定结果一致;最小检出量为1.57×104拷贝/ul。 相似文献
4.
根据GenBank收录的美洲型猪繁殖与呼吸综合征病毒（PRRSV）ATCCVR-2332株ORF6和ORF7基因序列，用O1igo软件设计并合成大小为37bp的寡核苷酸探针，经生物素标记后，成功建立了原住检测石蜡组织切片中PRRSV核酸的方法。该探针能检测到56PgPRRSV核酸的RT—PCR产物DNA，能特异检测出PRRSV核酸及其PCR产物，而对猪瘟病毒（HCV）、猪细小病毒（PPV）、猪伪狂犬病病毒（PRV）、猪乙脑病毒（JEV）的核酸呈阴性反应。应用该方法检测PRRSVSC-1株人工感染的28日龄仔猪，在感染后7d即可在肺脏、肾脏、扁桃体、胸腺、肺门淋巴结、十二指肠和大脑检测到PRRSV核酸。该法可用于仔猪PRRSV感染的诊断和组织中核酸的定位及分布研究，也可用于甲醛固定组织的回顾性诊断。 相似文献
5.
间接酶免疫组化检测DPV在感染鸭体内细胞定位的研究和应用 总被引：4，自引：3，他引：4
以超速离心浓缩结合蔗糖密度梯度离心法提纯的鸭瘟病毒（DPV）作为抗原免疫家兔制备兔抗DPV血清,用饱和硫酸铵沉淀结合DEAE-Sephedax-A-50离子交换柱层析提纯兔抗DPV IgG,建立了检测甲醛固定鸭体组织石蜡切片上DPV抗原的间接酶免疫组化法。该法能特异地检测出DPV感染鸭组织中的DPV而对鸭疫里默氏菌、鸭源多杀性巴氏杆菌（5∶A）、鸭源大肠杆菌（O1）感染致死的鸭组织以及正常鸭组织呈阴性反应,检测DPV感染死亡鸭的肝、十二指肠、脑、脾和胸腺的结果与PCR检测结果相同。其敏感性高于原位杂交。对DPV CH强毒株人工感染致死鸭的检测结果表明,该法可特异检测到肝、肺、肾、脑、十二指肠、空肠、回肠、直肠、法氏囊、脾脏、腺胃以及食管中的DPV。DPV主要分布于这些器官的上皮细胞和巨噬细胞。对1992-2004年经10%福尔马林保存的鸭瘟临床病例的肝脏检测结果均为阳性。该法可用于DPV感染鸭的诊断和定位检测,也可用于对甲醛固定组织进行回顾性诊断检测。 相似文献
6.
实时荧光定量PCR快速检测鸭病毒性肠炎标准方法的建立 总被引：1，自引：0，他引：1
根据GenBank(登录号:EF417996)中鸭病毒性肠炎病毒U31基因的序列,设计了1对特异性引物及TaqMan探针,扩增片段长度为76 bp.以鸭病毒性肠炎弱毒疫苗株DNA为阳性标准品模板,建立了实时荧光定量PCR检测鸭病毒性肠炎病毒的方法.该方法能在鸭病毒性肠炎病毒DNA样本中检出荧光信号,定量范围为:2.1×109～2.1×100个拷贝数,最小检出量为2.1×100个拷贝;用该方法对人工感染试验的4只鸭组织器官、粪便、血液等样品重复测定3次,病毒模板DNA的检出率为100%,对鸭正常组织、巴氏杆菌、大肠杆菌、沙门菌和鸭病毒性肝炎、鹅源禽流感H5毒株、新城疫、小鹅瘟病毒等DNA检测不出现特异性荧光信号.经过重复性和实际临床样本检验证实,该方法真实可靠,而且从核酸提取到报告检测结果耗时不超过4 h,不仅实现了对鸭病毒性肠炎的快速诊断,也实现了对该病毒DNA由定性到定量的检测. 相似文献
7.
从GenBank下载禽流感病毒(Auian in fluenza virus,AIV)NP基因序列,通过比对选取NP基因中较为保守的片段,设计1对引物,通过RT-PCR方法扩增270 bp的目的片段,经测序确认目的片段序列正确后,用地高辛标记扩增产物制备探针.在BSL-3实验室,用AIV人工感染鸡胚成纤维细胞和SPF鸡后,制作细胞涂片和组织石蜡切片,然后在经前处理后的细胞涂片和石蜡切片上进行原位RT-PCR,再与地高辛标记的探针进行原位杂交,对细胞和组织中的AIV进行检测和定位.研究结果表明,本方法能检测出约1μg/L质粒的DNA,并能特异性地在细胞涂片和石蜡组织切片中检测和定位AIV,且成纤维细胞感染病毒8 h后即可检测到阳性信号,具有良好的特异性和较高的敏感性,为动物组织中AIV的检测定位和致病机理研究提供了敏感、特异和更为直观的方法. 相似文献
8.
本研究建立了检测鸭瘟病毒（Duck pl ague vi rus,DPV）的PCR方法,并运用建立的检测方法对分离毒株和人工感染样品进行临床应用检测。根据GenBank（登录号为EF643558）中的DPV UL35基因保守区域,设计合成了一对引物,以DPV疫苗株为模板,优化PCR反应条件,建立了一种快速、有效的DPVPCR方法。结果显示：该方法能从DPV中扩增到与预期大小相符,长度为354 bp的特异性片段,而对禽流感病毒（AIV）、番鸭呼肠孤病毒（MDRV）、新城疫病毒（NDV）、番鸭细小病毒（MPV）、鹅细小病毒（GPV）等样品的扩增结果均为阴性;检测灵敏度达到470 ng病毒DNA。应用该方法对3株DPV分离株和6份由DPV BL8毒株人工感染鸭的肝脏和脾脏等组织进行PCR检测均为阳性。表明所建立的DPV PCR方法特异性强、灵敏度高,可用于DPV的临床诊断和流行病学调查。 相似文献
9.
10.
以猪繁殖与呼吸综合征 ( PRRS)病毒 ATCC VR-2 332株基因组为模板 ,通过 RT-PCR扩增出 586bp的 ORF6片段的 c DNA。将该片段克隆进 p GEM-T easy载体 ,转化大肠杆菌 JM1 0 9,成功获得重组质粒转化菌。将重组质粒经 Eco R 及 Sma 双酶切 ,回收到 1个 4 63bp的 c DNA片段 ,并制备出地高辛标记的c DNA探针。特异性检测表明 ,该探针对 PRRS病毒的 PCR产物呈现特异性 ,而对照的 HCV、PPV、PRV的核酸均呈阴性 ;敏感性检测表明 ,该探针对同源 DNA的检出限量为 4 pg。应用该探针对 6株 PRRS病毒北京分离毒及 ATCC VR-2 332株感染细胞进行原位杂交检测 ,结果均呈阳性。以上结果表明 ,该探针可成功用于组织原位杂交检测 相似文献
11.
Slocombe JO McMillan I 《The Canadian veterinary journal. La revue veterinaire canadienne》1984,25(9):347-350
In late December 1983, 2 800 veterinarians across Canada were sent a questionnaire in order to assess the status of heartworm disease in Canada in 1983 and 26% of them responded. Veterinarians reported that 59 504 dogs were blood-tested to check for microfilariae and 771 dogs (1.30% of those tested) were found with Dirofilaria immitis. Heartworm disease was diagnosed in all provinces except New Brunswick and Newfoundland but most (733) of the cases were in Ontario.
Heartworm disease was found most frequently in companion dogs over three years of age maintained mainly outdoors in rural areas. About 31% of the cases were observed with clinical signs of heartworm disease and 64% had a history of not having left Canada. Southwestern Ontario continues to be the focus of the infection and most of the dogs there had not left the province previously.
相似文献12.
Heartworm in dogs in Canada in 1982 总被引：8，自引：8，他引：0
Slocombe JO McMillan I 《The Canadian veterinary journal. La revue veterinaire canadienne》1983,24(7):227-229
13.
Owen J Slocombe D McMillan I 《The Canadian veterinary journal. La revue veterinaire canadienne》1986,27(9):324-328
In late December 1985, 1485 institutional veterinarians and small and mixed animal clinics across Canada were sent a questionnaire in order to assess the status of Dirofilaria immitis in Canada in 1985 and 44% of them responded. Veterinarians reported that 137,300 dogs were blood-tested to check for microfilariae and 1210 dogs were found with heartworm. Another 36 dogs were amicrofilaremic but diagnosed with heartworm disease to give the total number diagnosed in 1985 as 1247 (0.91%).
Heartworm was reported from all provinces except Prince Edward Island, Newfoundland and Saskatchewan but most (1126) of the cases were in Ontario. Southwestern Ontario continued to be the primary focus of the infection in Canada. From Quebec, 91 cases were reported mostly from and around Montreal. From Manitoba, 19 cases were reported from Winnipeg and surrounding areas. Heartworm was found most frequently in companion dogs over three years of age maintained mainly outdoors in rural areas. About 28% of the cases were observed with clinical signs of heartworm disease and 78% had a history of not having left Canada.
相似文献14.
Slocombe JO McMillan I 《The Canadian veterinary journal. La revue veterinaire canadienne》1989,30(6):504-508
In late November 1988, 1581 small and mixed animal clinics and institutional veterinarians across Canada were sent a questionnaire in order to assess the status of Dirofilaria immitis in Canada in 1988, and 46% of them responded. Veterinarians reported that 181,577 dogs were blood-tested for heartworm disease and 367 dogs were found with D. immitis microfilariae. Another 60 dogs were amicrofilaremic but diagnosed with heartworm disease to give the total number of cases diagnosed in 1988 as 441 (0.24%). 相似文献
15.
Slocombe JO McMillan I 《The Canadian veterinary journal. La revue veterinaire canadienne》1985,26(10):323-327
In late December 1984, 1853 institutional veterinarians and small and mixed animal clinics across Canada were sent a questionnaire in order to assess the status of Dirofilaria immitis in Canada in 1984 and 35% of them responded. Veterinarians reported that 97,794 dogs were blood-tested to check for microfilariae and 1417 dogs (1.45% of those tested) were found with heartworm. Another 34 dogs were amicrofilaremic, but were diagnosed as having heartworm disease, to give the total number diagnosed in 1984 as 1451 (1.48%). Heartworm was reported from all provinces except Prince Edward Island and Newfoundland but most (1310) of the cases were in Ontario. In Quebec, 126 cases were reported mostly from west of Montreal.
Heartworm was found most frequently in companion dogs over three years of age maintained mainly outdoors in rural areas. About 27% of the cases were observed with clinical signs of heartworm disease and 72% had a history of not having left Canada. Southwestern Ontario continued to be the primary focus of the infection.
相似文献16.
17.
Slocombe JO Villeneuve A 《The Canadian veterinary journal. La revue veterinaire canadienne》1993,34(10):630-633
In late November 1991, 1883 clinics in Canada were sent a questionnaire to assess the status of Dirofilaria immitis in dogs in 1991 and there was a 60.0% response. There were 344,031 dogs tested for heart-worm (HW), 627 were found infected and the prevalence of HW infection was 0.18%. There were 417 dogs with HW in Ontario, 116 in Manitoba, 38 in Quebec, 53 in British Columbia, three in Alberta, and one in Nova Scotia. In British Columbia, all of the infected dogs but one were from the Okanagan valley which, as from 1991, is a new focus of infection in Canada. Most dogs with HW had not been on preventive medication in 1990, and the prevalence among dogs tested and unprotected was 0.59%. That prevalence was considerably higher in endemic areas. Companion dogs, over three years of age and maintained primarily outdoors in rural areas, were most frequently infected. One cat was diagnosed with D. immitis and 33 dogs had Dipetalonema reconditium. 相似文献
18.
In late November 1989, 1732 clinics and institutional veterinarians were sent a questionnaire to assess the status of Dirofilaria immitis, and 51.7% responded. Of 247,716 dogs tested, 394 had D. immitis microfilariae and 51 were amicrofilaremic for a total of 445 cases and heartworm prevalence of 0.17%. Most (408) of these dogs had no preventive medication and the prevalence among dogs tested and unprotected was 1.01%. That prevalence was considerably higher in endemic areas. Thirty-seven dogs with heartworm had preventive medication. Heartworm was most frequent in companion dogs over three years of age maintained outdoors in rural areas. About 75% of the cases had never left Canada, 26% had clinical signs and 125 were not treated.
Heartworm was reported from British Columbia, Manitoba, Ontario, Quebec, Nova Scotia and Newfoundland, but 383 cases were in Ontario. South-western Ontario was the primary focus of infection. There were 33 cases in Quebec and 24 in Manitoba, mainly found in and around Metropolitan Montreal and Winnipeg respectively.
相似文献19.
Slocombe JO McMillan I 《The Canadian veterinary journal. La revue veterinaire canadienne》1988,29(8):641-646
In late November 1987, 1246 institutional veterinarians and small and mixed animal clinics across Canada were sent a questionnaire in order to assess the status of Dirofilaria immitis in Canada in 1987, and 50% of them responded. Veterinarians reported that 165,428 dogs were blood tested for heartworm disease and 511 dogs were found with D. immitis microfilariae. Another 78 dogs were amicrofilaremic but diagnosed with heartworm disease to give the total number of cases diagnosed in 1987 as 589 (0.35%). 相似文献
20.
Owen J Slocombe D McMillan I 《The Canadian veterinary journal. La revue veterinaire canadienne》1987,28(8):491-495
In late December 1986, 1224 institutional veterinarians and small and mixed animal clinics across Canada were sent a questionnaire in order to assess the status of Dirofilaria immitis in Canada in 1986; 46% of them responded. Veterinarians reported that 150,989 dogs were blood-tested for microfilariae and 869 dogs were found with heartworm. Another 65 dogs were amicrofilaremic but diagnosed with heartworm disease and one was found with heartworm at necropsy to give the total number diagnosed in 1986 as 935 (0.62%).
Heartworm was reported from Manitoba, New Brunswick, Ontario and Quebec, but most (810) of the cases were from Ontario. South-western Ontario continued to be the primary focus of the infection in Canada. There were 103 cases reported from Quebec, mostly from and around Montreal, and 21 cases from Manitoba, from Winnipeg and surrounding areas. Heartworm was found most frequently in companion dogs over three years of age maintained mainly outdoors in rural areas. About 33% of the cases were observed with clinical signs of heartworm disease and 81% had a history of not having left Canada.
相似文献