| 荷斯坦奶牛CN和DUMPS遗传缺陷病的巢式PCR检测方法的建立和应用 |
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| 引用本文： | 代元元 / 吴晓薇 / 洪洁心 / 等. 荷斯坦奶牛CN和DUMPS遗传缺陷病的巢式PCR检测方法的建立和应用[J]. 中国动物检疫, 2014, 0(1): 76-80,85 |
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| 作者姓名： | 代元元 / 吴晓薇 / 洪洁心 / 等 |
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| 作者单位： | [1]华南农业大学兽医学院,广东广州510642 [2]广东出入境检验检疫局,广东广州510623 [3]苏州科技学院,江苏苏州215009 |
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| 摘 要： | 为建立荷斯坦奶牛瓜氨酸血症和尿苷酸核酶缺乏症两种遗传缺陷病的检测方法，根据文献报道的精氨酸琥珀酸合成酶、UMPS基因核苷酸序列，设计引物，通过条件的优化，建立了巢式PCR反应体系。结合PCR产物测序，建立了检测CN和DUMPS方法。取243份临床牛血清样品用巢式PCR检测并测序，发现CN野生型238份，CN杂合子型5份，CN突变型0份；DUMPS野生型236份，DUMPS杂合子型5份，DUMPS突变型2份。所建立的巢式PCR方法灵敏、特异，还兼具高通量的特点，适合口岸对荷斯坦奶牛CN和DUMPS遗传缺陷病的大样品量检测。
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| 关 键 词： | 荷斯坦奶牛 瓜氨酸血症 尿苷酸核酶缺乏症 巢式PCR |
Development and Application of a Nested PCR Assay for Detection of CN and DUMPS in Holstein Cattle |
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| Dai Yuanyuan,Wu XiaoweiL,Hong Jiexin,Wang Yiffei,Liu Zhongyong,Zhao Tianchu,Guo Hui,Guo Xiaofeng. Development and Application of a Nested PCR Assay for Detection of CN and DUMPS in Holstein Cattle[J]. China Animal Health Inspection, 2014, 0(1): 76-80,85 |
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| Authors: | Dai Yuanyuan Wu XiaoweiL Hong Jiexin Wang Yiffei Liu Zhongyong Zhao Tianchu Guo Hui Guo Xiaofeng |
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| Affiliation: | 1.College of Veterinary Medicine, South China Agricultural University, Guangzhou 510642, China 2. Guangdong Entry- Exit Inspection and Quarantine Bureau, 510623; 3. Suzhou University of Science and Technology, Suzhow, Jiangsu 215009 ) |
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| Abstract: | The aim of the study was to develop a diagnostic method for detecting the two genetic defects---CN and DUMPS in Holstein cattle. Nested PCR primers were designed based on ASS gene and UMPS gene and PCR conditions were optimized. 243 cow blood samples were detected to confirm the presence of CN and DUMPS in the cows using the developed Nested PCR method and the PCR products were sequenced. The results showed that 238 cows were with-out CN, 5 suffered from heterozygote CN, and 236 cows without DUMPS, 5 heterozygote DUMPS and 2 mutant DUMPS respectively. These results indicated that the new assay was suitable to handle large quantity of CN and DUMPS deficient dairy cows in port quarantine inspection. |
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| Keywords: | Holstein cattle CN DUMPS nested PCR |
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