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本研究根据GenBank上发表的牛瑟氏泰勒虫18S rRNA基因的核苷酸序列设计并合成1对特异性引物,对寄生于牛体内的瑟氏泰勒虫基因组DNA进行扩增,得到1 356 bp的18S rRNA基因片段,测序后blast分析表明该虫种属牛瑟氏泰勒虫。将该基因片段序列与GenBank中8种已知泰勒虫的相应序列进行比较分析,建立系统发育树。结果表明,牛瑟氏泰勒虫吉林分离株与水牛泰勒虫亲缘关系最近,与小泰勒虫亲缘关系较远。这一结果说明宿主因素对泰勒虫的基因型影响较大。 相似文献
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牛中华泰勒虫新种(梨形虫亚目:泰勒虫科)2.分子分类学研究 总被引：6，自引：0，他引：6
分离自我国甘肃中部地区土种黄牛的一种形态特异的泰勒虫，采用传统分类学研究鉴定为新种，被定名为中华泰勒虫（Theileria sinensis sp.nov）。又利用对泰勒虫属特异的PCR引物（92/93）对该种基因组DNA扩增，结果表明该种属泰勒虫属原虫确定无疑，并对代表虫种进化和分类的18S rRNA基因序列进行了测定，对已测出该基因1067bp长片断序列与基因库中9种巴贝斯虫，7种已知牛泰勒虫的相应序列进行比较、分，建立起系统发生树。树图表明，瑟氏泰勒虫和水牛泰勒虫的亲缘关系非常近， 中华泰勒虫与这两个种的亲缘关系较近，与附膜泰勒虫、小泰勒虫、环形泰勒虫、斑羚泰勒虫、突变泰勒虫差异较大。 相似文献
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羊泰勒虫PCR检测方法的建立和初步应用 总被引：1，自引：0，他引：1
利用羊泰勒虫18SrRNA基因的序列特点,设计合成种特异性引物,建立羊泰勒虫PCR检测方法,该方法能特异性扩增398bp的羊泰勒虫18SrRNA基因片段,而对羊巴贝斯虫、羊无浆体、牛环形泰勒虫和牛伊氏锥虫的基因组DNA没有扩增带出现。对羊泰勒虫基因组DNA的最小检测量为0.12fgDNA。通过检测124份临床样品,24份为羊泰勒虫感染阳性,其余为阴性。结果表明,建立的PCR检测方法具有极高的敏感性和特异性,可用于羊泰勒虫病和临床健康带虫羊的诊断。 相似文献
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《中国兽医杂志》2017,(6)
为建立特异、敏感、快速的羊泰勒虫病诊断方法。根据Gen Bank已报道的羊泰勒虫表面蛋白基因(AY274329)设计合成了1对引物,通过条件优化,建立了羊泰勒虫病二温式PCR诊断方法。该方法能扩增出335 bp的羊泰勒虫特异性基因片段,测序结果与已知基因序列同源性为100%。对羊泰勒虫基因组DNA的最小检测量为16 fg/μL,与新孢子虫、弓形虫、巴贝斯虫和瑟氏泰勒虫均无交叉反应。与普通PCR相比,二温式-PCR在敏感性方面无显著差异,但其反复升温降温时间消耗短。结果表明,二温式PCR诊断方法具有较高的特异性和敏感性,可用于羊泰勒虫病的快速诊断和流行病学调查。 相似文献
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为筛选羊尤氏泰勒虫裂殖子功能基因,用羊尤氏泰勒虫裂殖子提取物免疫BALB/c小鼠,将小鼠脾脏细胞与鼠骨髓瘤SP2/0-Ag14细胞融合,融合细胞培养上清用酶联免疫吸附法和免疫印记法检测。提取羊尤氏泰勒虫裂殖子mRNA后,进行cDNA合成与文库构建。文库在免疫学噬斑筛选后,将阳性克隆进行测序和序列BLAST搜索。筛选结果显示有两个阳性克隆。所得序列BLAST搜索结果显示,它与12类寄生虫新生多态相关复合物α多肽基因的同源性在39%到78%之间。结果提示,所筛选的序列为羊尤氏泰勒虫裂殖子新生多态相关复合物α多肽基因不完全序列。该研究为今后使用单克隆抗体筛选羊泰勒虫功能性基因提供了参考。 相似文献
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为寻求一种快速灵敏的环形泰勒虫病PCR检测方法,基于环形泰勒虫裂殖体表面蛋白（Theirelia annulata surface protein,TaSP）基因序列保守区设计特异性引物,通过PCR技术扩增出该基因长为393 bp的高免疫原性区片段。用该引物对环形泰勒虫、中华泰勒虫、瑟式泰勒虫、尤氏泰勒虫、吕氏泰勒虫、绵羊泰勒虫、马泰勒虫、驽巴贝斯虫、牛巴贝斯虫基因组模板进行特异性试验,对环形泰勒虫基因组模板进行梯度稀释后扩增,以确定试验的敏感性,同时用本试验建立的方法与常规显微镜镜检方法对150份血清样品进行检测。特异性试验结果显示,在被检测的9个样本中,只有环形泰勒虫基因组模板中扩增出了符合大小的特异核苷酸片段;敏感性试验结果表明,PCR对环形泰勒虫的扩增效率可达到10-10;通过对150份血清样品的检测,并与血涂片方法进行比较,结果显示PCR方法具有特异性强、敏感度高等特点,适用于牛环形泰勒虫病的检测。 相似文献
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线粒体在真核生物以及单细胞生物的生命活动中发挥着重要作用。寄生虫为适应不同的生存条件进化了一套自己的方式,包括线粒体基因组成、基因表达和能量代谢方式。梨形虫（巴贝斯虫和泰勒虫）是一类重要的胞内寄生虫,给畜牧业造成重大的经济损失。本文对梨形虫为适应宿主而进化出的一些特征进行了描述,为寻找防治梨形虫的新途径提供理论依据。 相似文献
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本研究根据GenBank牛瑟氏泰勒虫P33表面蛋白基因设计1对引物,利用全血基因组DNA提取试剂盒提取牛瑟氏泰勒虫全血基因组DNA,PCR扩增得到了大小为786 bp的基因片断,测序分析证明,它与已报道序列的核苷酸同源性可达99%.将该基因与pET-28a(+)载体进行连接,经酶切和PCR鉴定,证明成功构建了含有目的基因片段的重组表达载体pET-28a-P33.将此质粒转化至大肠杆菌Rosetta(DE3)中,用IPTG进行诱导.结果证实,该基因在大肠杆菌中用IPTG诱导4 h时表达量达到高峰,表达产物为分子质量约30.3 ku的融合蛋白,其表达量占菌体总蛋白的43%,以包涵体形式存在;Western blotting试验证明,表达蛋白P33可被牛瑟氏泰勒虫阳性血清所识别,表明该融合蛋白具有较好的反应原性. 相似文献
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Only in recent years, the draft sequences for several agricultural animals have been assembled. Assembling an individual animal’s entire genome sequence or specific region(s) of interest is increasingly important for agricultural researchers to perform genetic comparisons between animals with different performance. We review the current status for several sequenced agricultural species and suggest that next generation sequencing (NGS) technology with decreased sequencing cost and increased speed of sequencing can benefit agricultural researchers. By taking advantage of advanced NGS technologies, genes and chromosomal regions that are more labile to the influence of environmental factors could be pinpointed. A more long term goal would be addressing the question of how animals respond at the molecular and cellular levels to different environmental models (e.g. nutrition). Upon revealing important genes and gene-environment interactions, the rate of genetic improvement can also be accelerated. It is clear that NGS technologies will be able to assist animal scientists to efficiently raise animals and to better prevent infectious diseases so that overall costs of animal production can be decreased. 相似文献
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The first large-scale urban survey of Giardia infections in dogs was undertaken in the USA. It involved several locations in the Western United States with Giardia isolates from microscopy-positive samples characterised by multi-locus PCR and sequencing. A high prevalence of Giardia was confirmed in asymptomatic domestic dogs, and for the first time, provides evidence that zoonotic assemblages/subgroups of Giardia occur frequently in domestic dogs living in urban environments, and more frequently than the dog specific assemblages. 相似文献
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Beth Harris Jessica Hicks Melanie Prarat Susan Sanchez Beate Crossley 《Journal of veterinary diagnostic investigation》2021,33(2):248
With the cost of next-generation sequencing (NGS) decreasing, this technology is rapidly being integrated into the workflows of veterinary clinical and diagnostic laboratories nationwide. The mission of the U.S. Department of Agriculture–National Animal Health Laboratory Network (NAHLN) is in part to evaluate new technologies and develop standardized processes for deploying these technologies to network laboratories for improving detection and response to emerging and foreign animal diseases. Thus, in 2018, the NAHLN identified the integration of NGS into the network as a top priority. In order to assess the current state of preparedness across NAHLN laboratories and to identify which have the capability for performing NGS, a questionnaire was developed by the NAHLN Methods Technical Working Group and submitted to all NAHLN laboratories in December 2018. Thirty of 59 laboratories completed the questionnaire, of which 18 (60%) reported having some sequencing capability. Multiple sequencing platforms and reagents were identified, and limited standardized quality control parameters were reported. Our results confirm that NGS capacity is available within the NAHLN, but several gaps remain. Gaps include not having sufficient personnel trained in bioinformatics and data interpretation, lack of standardized methods and equipment, and maintenance of sufficient computing capacity to meet the growing demand for this technology. 相似文献
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Staphylococcus simulans, a coagulase-negative staphylococcal species, can cause several diseases in humans and animals. This report describes a case of dermatosis characterized by broad, well-circumscribed hyperkeratosis and alopecia on the back of a household pygmy hedgehog (Erinaceous albiventris). Quills and exudates were studied by microscopy. The microscopic examination of the exudates collected from the lesion revealed several leukocytes and numerous gram-positive cocci. An aerobic bacterial culture revealed overgrowth of the same gram-positive bacteria. The bacterium was identified as S. simulans by polymerase chain reaction amplification and direct sequencing targeted to the 16S ribosomal RNA gene. This report is the first to show that S. simulans could be related to the dermatitis of hedgehogs. 相似文献
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DNA测序技术从以Sanger法为代表的第一代测序技术到如今的高通量测序(HTS)技术,经历了漫长而曲折的发展过程,如今该技术在动植物基因组的研究中获得了很大的成功,它的问世可以使人们以更低廉的价格,更全面、更深入地对全基因组进行分析.作者阐述了几种关于HTS的测序技术平台:Roche/454测序技术、ABI/SOLID测序技术、Illumina/Solexa测序技术、单分子测序技术及Ion Torrent测序技术,并且还归纳了HTS在基因组学、转录组学及表观基因组学等方面的应用. 相似文献
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Imputation from SNP chip to sequence: a case study in a Chinese indigenous chicken population 总被引：1，自引：0，他引：1
Shaopan Ye Xiaolong Yuan Xiran Lin Ning Gao Yuanyu Luo Zanmou Chen Jiaqi Li Xiquan Zhang Zhe Zhang 《畜牧与生物技术杂志(英文版)》2018,(2)
Background: Genome-wide association studies and genomic predictions are thought to be optimized by using whole-genome sequence(WGS) data. However, sequencing thousands of individuals of interest is expensive.Imputation from SNP panels to WGS data is an attractive and less expensive approach to obtain WGS data. The aims of this study were to investigate the accuracy of imputation and to provide insight into the design and execution of genotype imputation.Results: We genotyped 450 chickens with a 600 K SNP array, and sequenced 24 key individuals by whole genome re-sequencing. Accuracy of imputation from putative 60 K and 600 K array data to WGS data was 0.620 and 0.812 for Beagle, and 0.810 and 0.914 for FImpute, respectively. By increasing the sequencing cost from 24 X to 144 X, the imputation accuracy increased from 0.525 to 0.698 for Beagle and from 0.654 to 0.823 for FImpute. With fixed sequence depth(12 X), increasing the number of sequenced animals from 1 to 24, improved accuracy from 0.421 to0.897 for FImpute and from 0.396 to 0.777 for Beagle. Using optimally selected key individuals resulted in a higher imputation accuracy compared with using randomly selected individuals as a reference population for resequencing. With fixed reference population size(24), imputation accuracy increased from 0.654 to 0.875 for FImpute and from 0.512 to 0.762 for Beagle as the sequencing depth increased from 1 X to 12 X. With a given total cost of genotyping, accuracy increased with the size of the reference population for FImpute, but the pattern was not valid for Beagle, which showed the highest accuracy at six fold coverage for the scenarios used in this study.Conclusions: In conclusion, we comprehensively investigated the impacts of several key factors on genotype imputation. Generally, increasing sequencing cost gave a higher imputation accuracy. But with a fixed sequencing cost, the optimal imputation enhance the performance of WGP and GWAS. An optimal imputation strategy should take size of reference population, imputation algorithms, marker density, and population structure of the target population and methods to select key individuals into consideration comprehensively. This work sheds additional light on how to design and execute genotype imputation for livestock populations. 相似文献
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Chiers K Deschaght P De Baere T Dabrowski S Kotlowski R De Clercq D Ducatelle R Vaneechoutte M 《Comparative immunology, microbiology and infectious diseases》2012,35(4):303-307
Routine cultivation methods are able to distinguish between isolates of the Mycobacterium avium and the Mycobacterium tuberculosis complex. However, molecular tools are needed to further identify the several subspecies in the M. avium complex, especially for the subspecies avium and silvaticum. A rapid technique using HhaI restriction digestion of a 349 bp amplification product of the 85B antigen (α-antigen) gene was used for the identification of M. avium subsp. silvaticum in a three-year-old gelding presenting with caseous, necrotizing, granulomatous lesions. The result was confirmed by sequencing of the 85B antigen gene. 相似文献
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基因组测序技术是一种分子生物学及其相关学科研究中最常用的技术。从20世纪70年代中期第一代测序技术出现以来,基因组测序技术已取得了重大进展,并改变了生命科学诸多领域的研究面貌。测序成本的急剧下降,测序通量呈指数提高,使得测序速度大大提升。DNA和RNA是生命体的2个基本遗传物质,其组成和序列变化创造了形形色色的生命世界。快速、准确地获取生物体的遗传信息对于生命科学的研究具有重要意义,测序技术能够真正地反映基因组、遗传信息转录,全面地揭示基因组的复杂性和多样性,在生命科学研究中起着重要的作用。综述了各代测序技术的发展成果、测序原理、测序优缺点、国内研究现状,并对测序技术未来的发展方向进行了展望。 相似文献