Flow cytometry: clinical applications in equine medicine     

Davis EG  Wilkerson MJ  Rush BR 《Journal of veterinary internal medicine / American College of Veterinary Internal Medicine》2002,16(4):404-410

The use of flow cytometry in veterinary diagnostics is becoming a valuable clinical tool with a broad range of applications. Physical characteristics of cells can be determined by the flow cytometer laser and electronics through the measurement of changes in light scatter properties. Other components and functions of cells can be defined through the application of fluorochrome dyes that have an affinity for cellular components. Traditionally, common clinical applications are immunophenotyping of cells of the hematopoietic system with fluorescent-labeled antibodies raised against specific cell surface proteins. Other approaches have been used to elucidate changes in cell function and DNA content. This review is intended to provide the reader with the fundamental uses of flow cytometry. Examples of clinical applications in equine patients include immune-mediated hemolytic anemia, immune-mediated thrombocytopenia (IMT), chronic inflammatory disease, and neoplasia.  相似文献   

Evaluation of the Abbott LCx Mycobacterium Tuberculosis Assay for Direct Detection of Mycobacterium Bovis in Bovine Tissue Samples     

Valente C  Cuteri V  Ausili E  Piersimoni C 《Veterinary research communications》2002,26(1):21-27

The commercial LCx amplification assay, usually employed to detect the Myocobacterium tuberculosis complex in respiratory specimens, was evaluated by comparing the results it gave with those obtained using Löwenstein-Jensen solid medium and pathological findings on 55 lymph nodes from cattle with positive and 10 lymph nodes from cattle with negative skin tests for tuberculosis. Fifty-three cultures (51 and 2, respectively) were positive for M. bovis, while the results for the LCx assay and the histological method were positive in 48 (45, 3) and 24 (20, 4) samples, respectively. None of the samples from cattle from certified tuberculosis-free herds were positive by any of the procedures. The results obtained with the LCx assay, compared with the culture procedure, regarded as the gold standard among the diagnostic techniques, gave a specificity of 91.6% and sensitivity of 90.5%. Although the sensitivity of LCx was suboptimal, DNA of M. bovis was detected in 81.8% of the skin test-positive animals. Amplification techniques could provide a rapid and reasonably reliable tool for detecting bovine tuberculosis.  相似文献   

DNA fingerprinting of Australian isolates of Mycobacterium avium subsp paratuberculosis using IS900 RFLP     

Cousins DV  Williams SN  Hope A  Eamens GJ 《Australian veterinary journal》2000,78(3):184-190

OBJECTIVES: To evaluate additional restriction enzymes for IS900 RFLP of Mycobacterium avium subsp paratuberculosis and examine the genetic diversity among Australian isolates for epidemiological studies of Johne's disease. DESIGN AND PROCEDURE: Seventy-one isolates of M paratuberculosis from cattle, sheep, goat, alpaca and rhinoceros in six Australian States and the Northern Territory, reference strains and reference DNA from previously characterised strains were tested for genetic variation. Bst EII, Pvu II and Pst I restriction enzymes were used, and four others (Bam HI, Alu I, Xho I and Dra I) were assessed for their ability to detect polymorphisms. Multiple isolates from some animals were tested. RESULTS: Bam HI, was the most effective enzyme for identifying polymorphisms (12 types), followed by Bst EII (11 types). Both Pvu II and Pst I were relatively ineffectual. Fifteen different types were identified, 12 in clinical isolates. Most isolates were cattle (C) strains and fell into the C1 (n = 28) and C3 (n = 32) groupings. All isolates from alpaca were type C1, and bovine isolates were commonly C1 (n = 15) or C3 (n = 28). All of the sheep were infected with sheep (S) strains; no S strains were identified in cattle. Two of six isolates from one animal had single band differences. CONCLUSION: The epidemiological features of M paratuberculosis in Australia are similar to those reported in New Zealand, where cattle and sheep are commonly infected with different strains. However, because of the lack of polymorphism identified within the major groups, it is unlikely that DNA fingerprinting will have a significant role in epidemiological studies of Johne's disease, unless an unusual strain in being studied.  相似文献   

扁蓿豆遗传多样性研究初探     

李鸿雁  米福贵  李志勇  师文贵  李兴酉 《四川草原》2007,(1)

论述了扁蓿豆遗传多样性研究的依据,利用不同生境条件下生长的野生和栽培品种,采用形态标记、细胞标记、生化标记和分子标记的方法多方面地对扁蓿豆进行遗传多样性的研究。  相似文献   

新扬州鸡早期增重的OPAY02-SCAR分子标记研究     

戴国俊  谢恺舟  王志跃  吴圣龙  施会强  Olowofeso O  盛浩伟  王金玉 《畜牧兽医学报》2006,37(6):544-549

OPAY02型2条多态性条带经克隆、测序和引物设计后,转换成SCAR标记,并对86个新扬州鸡随机交配后代基因组DNA进行了PCR扩增.2条带DNA序列与红色原鸡基因组序列比对结果表明,大分子量条带与位于红色原鸡第3号染色体上序列有98%的同源性,共检测到8个SNPS,其中195位的碱基T→G,316位的A→T,538位的G→A,731位的T→A,1 147位的G→A,1 329位的T→C,1 927位的C→T,2 081位的C→T,小分子量条带与红色原鸡没有同源序列,推测新扬州鸡野祖除红色原鸡外,还有其它来源.SCAR标记分析表明,经条件优化随机扩增的OPAY02型标记稳定、可靠,可用于遗传分析.2条带所在座位群体基因型平衡性测验结果表明,所测新扬州鸡群体处于平衡状态,选择可以打破平衡,有利于动物育种.  相似文献   

牛基因组DNA两种提取方法的比较研究   总被引：3，自引：0，他引：3  

程金华  朱化彬  王栋  郝海生  杨波 《畜牧兽医学报》2006,37(9):874-877

研究哺乳动物基因组DNA的水煮抽提法，并将传统的酚仿抽提法和水煮法进行比较，水煮法即通过对细胞的煮沸和冷却，使细胞破裂、蛋白变性，从而获得用于PCR扩增的模板DNA的方法。通过电泳分析和PCR扩增检测了所提取基因组DNA的完整性、可行性。检测时采用3对引物对2种方法提取的DNA进行了PCR扩增，同时对冷冻保存的基因组DNA也进行了扩增检测。结果表明：水煮法提取基因组DNA是一种快速、简便、经济、高效的方法。  相似文献   

A-FABP 在不同鸡种中遗传多态性分析   总被引：6，自引：0，他引：6  

陈宽维  章双杰  屠云洁  汤青萍  高玉时  李慧芳 《畜牧兽医学报》2006,37(11):1114-1117

利用PCR—RFLP技术和DNA测序检测了尤溪麻鸡、鹿苑鸡、隐性白羽鸡3个鸡种共计96只鸡的脂肪细胞型脂肪酸结合蛋白（A—FABP）第85位点和第1805位点的遗传变异。结果表明：1）在这2个位点上，3个鸡种均存在变异，外来品种隐性白羽鸡以杂合子Cc居多；地方鸡种以cc居多，3个鸡种均含有较高频率的等位基因c，其中以尤溪麻鸡最高，而以隐性白羽鸡最低。2）A—FABP第85位点不影响A—FABP的氨基酸序列；而第1805位点的遗传多态性影响氨基酸序列的变化：由脯氨酸变为丝氨酸，说明该多态性可能通过A-FABP基因的表达水平从而影响鸡的肉质。本试验为进一步分析A-FABP基因的遗传变异与肌内脂肪含量的关系及该分子标记在育种计划中的应用奠定基础。  相似文献   

羊驼Cytb基因序列的测定及其研究   总被引：3，自引：0，他引：3  

董常生  高莉  贺俊平  赫晓燕  耿建军  赵英虎 《畜牧兽医学报》2006,37(12):1254-1258

测定了羊驼线粒体DNA细胞色素b基因部分序列，并与骆驼科其它物种细胞色素b基因进行同源序列比较，分析了碱基组成及变异情况，并用邻接法和最大简约法构建了分子系统树，在分子水平上探讨了羊驼在骆驼科动物中的分类地位。结果表明，羊驼与骆马和美洲驼存在的遗传差异相对比羊驼与原驼大，为传统分类学中的羊驼是原驼驯化以后的一种家养驼的提法提供了分子学依据。  相似文献   

从牛奶中分离DNA方法的建立   总被引：7，自引：0，他引：7  

田雨 《乳业科学与技术》2006,28(3):112-113

本文通过将牛奶中提取白细胞技术和热裂解提取DNA技术相结合建立了从牛奶中提取DNA的技术程序。结果显示，提取DNA原液的平均浓度为0．5μg／mL。通过PCR分析发现，提取DNA的浓度已达到PCR反应的模板要求，可以作为一种快速的DNA提取方法。  相似文献   

瘤胃微生物总DNA快速提取法研究     

淡瑞芳  龙瑞军  杨予海 《家畜生态》2006,(1)

本文介绍了一种简便快速地提取瘤胃微生物细胞总DNA的方法,该法是对提取植物的CTAB法进行改进而成。与经典反复冻融 SDS/蛋白酶K裂解法相比,该法可在数小时内高效地提取瘤胃内容物总DNA,经琼脂糖凝胶电泳对所提取瘤胃微生物总DNA进行检测,证明所获得的瘤胃总DNA符合分子生物学操作要求。  相似文献