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1.
2019年9月,陕西西安市某海兰褐蛋鸡场场出现产蛋率下降,死淘率升高,剖检部分死亡鸡只表现肝脾肿大且易脆破裂,为弄清该鸡场疫情的病因。我们采集了鸡血清32份,胆汁13份,粪便、肝脏和脾脏各11份,然后利用间接ELISA检测血清中是否存在禽HEV抗体,巢式RT-PCR检测粪便、肝脏、脾脏和胆汁样品中是否存在禽HEV核酸。间接ELISA检测结果发现,28份鸡血清禽HEV抗体阳性,阳性率为87.5%;巢式RT-PCR检测结果发现,6份胆汁,5份粪便,3份肝脏和脾脏样品为禽HEV RNA阳性。阳性PCR产物测序后基因序列同源性分析发现,获得的禽HEV ORF2部分基因序列相互之间同源性为99%～100%,与GenBank已知的禽HEV序列同源性在78%～99%,进化树分析表明分离的禽HEV与国内多数禽HEV分离株以及欧洲部分分离株在同一分支上,属于基因3型。上述临床剖检病变和实验室检测技术说明,该鸡场存在禽HEV的感染,此次疫情可能主要是由该病毒感染引起的。 相似文献
2.
为弄清辽宁某蛋鸡场鸡只产蛋量突然下降,死亡率上升及部分剖检鸡只肝脏肿大出血的原因,对采集的发病鸡肝脏组织进行病理学观察,同时对肠道内容物进行禽戊型肝炎病毒(Hepatitis E virus,HEV)核酸片段的RT-PCR检测。肝脏的组织病理学观察发现,肝细胞坏死和汇管区周围淋巴细胞浸润;RT-PCR从肠道内容物中成功扩增到禽HEV ORF2基因的部分片段;基因序列同源性分析结果表明,其与国内禽HEV参考株的序列同源性为97.5%~99.6%,而与其他国家的HEV序列同源性为77.9%~97.6%;进化树分析表明,该序列(命名为China 15-LN)与国内其他地区及欧洲的部分分离株在同一进化分支上,同属禽HEV基因3型。 相似文献
3.
为从核酸水平证实我国鸡群中禽戊型肝炎病毒(HEV)的存在,本实验从山东省某鸡场患有肝脾肿大综合征的病鸡中采集10份粪便和8份胆汁样品,利用RT-PCR方法检测其中禽HEV ORF2基因片段,并将阳性PCR产物克隆测序。结果显示:18份粪便和胆汁样品中,13份为禽HEV RNA阳性;其序列间的同源性为97%～99%,与GenBank中登录的参考序列同源性为76.6%～98.1%;进化树显示与欧洲地区的禽HEV在同一分支,属于禽HEV基因3型。禽HEV ORF2基因的检出为进一步了解禽HEV对我国家禽养殖业的危害以及在我国的流行情况奠定了基础。 相似文献
5.
为了解云南省猪群戊型肝炎感染情况,从云南省8个猪场采集166份新鲜猪粪便样品,用套式RT-PCR技术对戊型肝炎病毒(HEV)核酸进行检测,结果 21份样品阳性,阳性率为12.7%,感染率较高。通过PCR产物片段的测序分析,去除重复序列后得到11株基因序列。同源性分析结果表明,这11株毒株序列与选定的Gen Bank中基因1型、2型、3型和4型参考序列的核苷酸序列同源性分别为85.6%~87.8%,85.6%~87.8%,85.6%~95.6%和92.8%~98.9%,说明这些株毒株属于基因4型,与中国其他地区分离的毒株同源性最高,证实HEV的流行具有地理分布的特征。 相似文献
6.
为了解广西猪群中戊型肝炎病毒(HEV)基因型,利用套式RT-PCR用对南宁周边猪场采集到的104份新鲜猪粪便进行HEV检测、分离并成功扩增和克隆了11株阳性毒株的ORF2基因部分保守片段.序列测定结果表明,11株HEV ORF2序列自身核苷酸同源性为97.7%~100%,推导编码的氨基酸同源性为97.9%~100%;与4型HEV代表毒株Ch-S-1、Ch-T11、swGX40等的核苷酸同源性为84.6%~90.6%,而与其他各型之间的同源性较低.系统发育进化树结果表明,11株HEV广西株为基因4型,其中,10株HEV为4a亚型,1株毒株为4e亚型.结果表明,广西猪群中流行的HEV均为基因4型,且以4a亚型为优势流行毒株. 相似文献
7.
为了解河南地区猪戊型肝炎感染情况,同时掌握该地区猪感染戊型肝炎病毒的基因型,对来自河南新乡、平顶山、南阳等地区不同商品猪场血清、肝脏、粪样品,采用ELISA技术检测猪血清中抗HEV特异性抗体水平,并使用套式RT-PCR检测肝脏和粪便中猪戊型肝炎病毒核酸。结果显示530份样品中HEV抗体阳性率0.6%,HEV RNA均为阴性,推测目前河南省猪戊型肝炎病毒感染率较低。 相似文献
8.
采集江西省A猪场2～3月龄猪粪便40份,B猪场2～3月龄猪粪便40份,利用反转录套式聚合酶链方法(RT-nPCR),检测戊型肝炎病毒(hepatitis E virus,HEV)RNA,结果表明,A猪场7份样品为阳性,阳性率17.5%.B猪场10份样品为阳性,阳性率25.0%.总阳性率21.3%.对4份PCR扩增阳性产物进行纯化并测序,利用生物学软件进行序列分析和进化树绘制,4个分离株ORF2 348 bp同源性为92.1%～97.5%,为同一基因型,与HEV1、HEV2、HEV3同源性分别为70.7%～77.4%、72.5%~75.7%和71.6%～76.8%,与HEV4型的同源性79.2%～83.9%.4个分离株与HEV4型的代表株T1在同一分支上,属基因4型.与中国大陆6株猪源HEV比较,同源性为79.0%～87.3%,提示中国大陆猪源HEV的基因型比较一致,同属HEV 4型. 相似文献
9.
为了表达基因4型戊型肝炎病毒(HEV)上海株ORF2编码蛋白并分析其抗原性,采用套式RT-PCR方法扩增上海猪基因4型HEV代表株结构蛋白基因ORF2部分片段,构建含有该基因片段的pET30a(+)重组表达质粒,命名为pET-E;将重组表达质粒转化大肠杆菌BL21,经1mmol/L IPTG诱导得到表达,进行SDS-PAGE分析,然后用Western blot鉴定其抗原性,最后纯化蛋白。结果:SDS-PAGE证明该片段得到融合表达,分子量为33 ku。Western blot分析表明,重组蛋白可以与HEV阳性血清反应,表明该蛋白具有良好的抗原性;该融合蛋白含有组氨酸标签,可以用MagExtractor-His-tag-kit进行纯化。用纯化的重组蛋白作为包被抗原建立的间接ELISA检测89份猪血清样品,阳性率达80.4%,与已有的试剂盒检测结果相比,阳性符合率达95%。 相似文献
10.
戊型肝炎病毒(Hepatitis E virus,HEV)主要引起人的戊型肝炎,新近研究发现猪在病毒传播中可能发挥重要作用.本研究对我国部分省区HEV感染血清学调查,在被检的1 138份血清中,有666份(57.5%)为HEV抗体阳性,猪群抗体阳性率随着月龄增长而升高.通过RT-PCR方法从一份猪粪中扩增并克隆了HEVORF2 N端主要抗原决定区339bp基因片段,序列分析显示,该段基因与我国人群HEV基因4型毒株核苷酸序列同源性为85.9%,但氨基酸序列完全一致.这一结果提示我国猪群存在广泛的HEV感染,并在一定程度上与人群HEV毒株密切相关. 相似文献
11.
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
12.
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
13.
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
15.
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
16.
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
17.
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
19.
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
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