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将隐孢子虫鼠基因型(Cryptosporidium mouse genotype)P23基因亚克隆到pGEX-4T-1载体中,并用大肠杆菌BL21(DE3)为宿主菌诱导表达。以纯化的rP23为诊断抗原,建立隐孢子虫间接ELISA检测方法,观察其敏感性、特异性和重复性,并对临床血清样品进行检测。结果隐孢子虫鼠基因型P23基因在大肠杆菌中获得了高效表达,Western blot检测显示rP23能被隐孢子虫感染兔血清识别。以纯化rP23为诊断抗原,成功地建立了检测兔隐孢子虫病的间接ELISA技术。对23份临床血清检测结果显示,该方法检出率高于Sheather's蔗糖漂浮法、套式PCR法。本研究结果为研制兔隐孢子虫病诊断试剂盒打下了基础。 相似文献
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隐孢子虫病(Cryptosporidiosis)是一种全球性的人兽共患原虫病,具有广泛的宿主范围,可以感染鸟类、哺乳类、鱼类、爬行类、两栖类以及人在内的240多种动物。野生动物隐孢子虫做为人隐孢子虫病感染的重要传播来源,对其进行生物学研究具有重要的公共卫生意义。本文分别叙述了野生动物隐孢子虫的种类及基因型,为进一步研究野生动物的隐孢子虫病提供了有效的参考。 相似文献
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为将微小隐孢子虫(Cryptosporidium parvum)P23基因在巴斯德毕赤酵母(Pichia pastoris)系统中进行表达,利用表达蛋白初步建立隐孢子虫病间接ELISA诊断技术,设计引物从微小隐孢子虫基因组DNA中扩增P23基因序列,构建pPIC9K-P23重组质粒,在毕赤酵母中进行表达,用阴离子交换层析柱进行纯化。以重组P23纯化蛋白为抗原建立间接ELISA检测方法,对现场采集的猪血清样品进行检测。SDS-PAGE显示所表达的蛋白大小约为23 kDa。Western blot检测表明该蛋白能与兔抗P23蛋白血清特异性结合。用建立的间接ELISA技术对186份猪血清样品进行检测,阳性率为83.3%。本研究获得了真核表达的P23重组蛋白,初步建立了微小隐孢子虫病间接ELISA诊断技术,为隐孢子虫病的诊断和流行病学调查打下了基础。 相似文献
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禽隐孢子虫病是由火鸡隐孢子虫(Cryptosporidium meleagridis)和贝氏隐孢子虫(C、baileyi)引起的以消化道和呼吸道症状为主的禽类寄生原虫病。自1929年Tyzzer首次报道在鸡的盲肠上皮细胞上发现隐孢子虫以未,有关学者对禽隐孢子虫病的病原学、流行病学、临床病理学、诊断学等方面做了大量研究,取得了一定的成就。本文就禽隐孢子虫病的研究概况,作一综述。 相似文献
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应用巢式聚合酶链反应(Nested PCR)建立了一种检测隐孢子虫(Cryptosporidium)的方法。试验中隐孢子虫卵囊纯化采用庶糖密度梯度离心法,以液氮-热水浴反复冻融及酚-氯仿抽提冷乙醇沉淀法制备模板DNA,根据隐孢子虫18S rRNA序列高度保守区设计2对引物,建立Nested PCR诊断方法。该方法特异性强,可检出牛源微小隐孢子虫(C.parvum)、羊源微小隐孢子虫(C.parvum)、牛源安氏隐孢子虫(C.andersoni)、鸡源贝氏隐孢子虫(C.baileyi)及猪源隐孢子虫(C.suis);敏感性高,该方法最低核酸DNA检测量达到10fg。初步应用结果表明,所建立的Nested PCR方法适合于隐孢子虫病的诊断和分子流行病学调查。 相似文献
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隐孢子虫是可以感染小反刍动物的肠道原生生物,具有潜在的公共卫生学问题。隐孢子虫可引起人和动物的腹泻和肠道疾病,严重情况下会出现死亡。被感染的动物可能是人畜共患型隐孢子虫的宿主,可引起公共卫生风险、农场利润减少和动物福利等问题。隐孢子虫病已被报道是新生牛、羊等反刍动物腹泻和死亡的一个重要原因,认为是新生羔羊腹泻的第二大诱因,仅次于轮状病毒。羊隐孢子虫病在世界范围内都有发现,不同程度地威胁着人和动物的身体健康,基于此,现就隐孢子虫对羊感染的危险因素和常用检测方法,以及我国羊感染的现状进行综述,为其防控提供参考。 相似文献
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隐孢子虫病是由隐孢子虫(Cryptosporidium)寄生于人类、家养动物及多种野生动物胃肠道引起的一种严重的人畜共患胃肠疾病。该病不但威胁人类健康,亦严重影响畜牧业发展。近年来随着研究的深入,隐孢子虫的流行表现出许多新的特点,危害远远超出了人们的估计。这些特点主要表现在:感染家畜,增加人类感染风险;与动物其他病原混合感染危害加重;感染野生动物,具有自然疫源性;感染海洋和水生动物,造成水体污染;经水源和食物传播引起人类群体感染;暴发感染对人群危害严重。深入开展隐孢子虫病流行病及防控研究对提高公共卫生水平具有重要意义。 相似文献
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Information concerning histologic significance of naturally occurring bursal Cryptosporidium sp. infection in chickens was obtained by retrospective examination of histopathology reports and clinical data from histology accessions received during 1986. Cryptosporidiosis was diagnosed in 197 bursas. In two-thirds of the accessions, more than 50% of the examined bursas were infected with Cryptosporidium sp. The histologic morphologic lesion diagnosis for Cryptosporidium sp.-infected bursas most often was marked diffuse chronic-active superficial purulent protozoal bursitis with mucosal epithelial hyperplasia. Our study clearly indicates that Cryptosporidium sp. is associated with inflammation and disturbed growth in chicken bursas. Additionally, our data indicate that Cryptosporidium sp. infection is not dependent on bursal damage attributable to other agents, including infectious bursal disease virus. 相似文献
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C.Z. Chako J.W. Tyler L.G. Schultz L. Chiguma B.T. Beerntsen 《Journal of veterinary internal medicine / American College of Veterinary Internal Medicine》2010,24(1):37-43
Cryptosporidiosis is one of the most common causes of infectious diarrhea in people. Although dairy calves are high-risk hosts, the role of other livestock, pets, and humans in the disease should not be underestimated. Some Cryptosporidium species and strains are specific to people, others are specific to animals while some are zoonotic pathogens. Cryptosporidium hominis is the species responsible for the majority of human cases in the United States, Sub-Saharan Africa, and Asia, while Cryptosporidium parvum accounts for more human cases in Europe and particularly in the United Kingdom. A deeper understanding of Cryptosporidium host range, reservoirs, and transmission is needed to develop preventive strategies to protect the general public. 相似文献
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Miller DL Mauel MJ Liggett A Hines ME Frazier KS Pence M Whittington L Baldwin CA 《Veterinary journal (London, England : 1997)》2006,171(3):478-482
Although Cryptosporidium spp. are found throughout the world and in multiple environmental conditions, few data are available that explore the possibility of an association between specific environmental parameters and the species or strain of Cryptosporidium. This study examines the potential association between a particular Cryptosporidium species/strain found in calves and soil provinces in Georgia, USA. Necropsy cases spanning the years 1996-2002 were tested. No significant differences (P=0.962, chi(2) test of homogeneity) between numbers of positive cases were noted among soil provinces. Phylogenetic analysis of the sequences for the PCR products revealed sequence similarity of the products with Cryptosporidium parvum strain C1. Although, clinical Cryptosporidiosis in calves was not found to be affected by soil province and may be caused by a single genotype, other genotypes may be responsible for subclinical infection and warrant further investigation. 相似文献
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为快速检测并准确鉴别奶牛隐孢子虫种,以隐孢子虫18S rRNA基因的特殊区域为基础,设计内、外引物,并根据软件分析确定相应的内切酶EcoT141,采用Nested PCR-RFLP方法进行虫种种型鉴别分析。在Nested PCR两次PCR反应中,以微小隐孢子虫(Cryptos poridium parvum,C.p)和安氏隐孢子虫(Cryptosporidium andersoni,C.an)卵囊提取的DNA为模板,均能扩增出长约800bp和500bp的明亮条带,且特异性强,其他虫种不能扩增出条带,该方法最低可检测到5个卵囊/g粪便;对于由内引物扩增出的500bp的条带,C.an的PCR产物能被内切酶EcoT141酶切,酶切后的片段分别为416bp和92bp,C.p的PCR产物不能被此酶酶切。用所建立的Nested PCR-RFLP法对上海奶牛389头和进口奶牛200头的共计589份粪样进行检测,Nested PCR的结果表明上海奶牛和进口奶牛的隐孢子虫阳性率分别为19.02%和3.5%,RFLP的结果表明上海奶牛感染的主要是Cp和C.an,进口奶牛感染的主要是C.p。研究结果表明,本研究建立的检测奶牛粪便中的隐孢子虫的NestedPCR-RFLP法,可用于奶牛隐孢子虫流行病学调查并有效鉴别奶牛隐孢子虫种。 相似文献
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应用巢式聚合酶链反应(Nested PCR)-限制片段长度多态性 (restriction fragment length polymorphism, RFLP)方法对微小隐孢子虫(C.parvum)、安氏隐孢子虫(C.andersoni)和火鸡隐孢子虫(C.meleagrides)的鉴别进行了研究。结果显示C.parvum BOCC2、C.andesoni BOCC2和C.meleagrides CHCC1扩增产物片段大小分别为830bp、828bp和828bp,扩增产物分别经VspI酶切后形成3种不同的RFLP图谱,根据RFLP图谱可鉴别C.parvum、C.andersoni和C.meleagrides。本研究为我国隐孢子虫的分类和隐孢子虫病的分子流行病学研究打下了良好基础。 相似文献