研究报告 - 中国实验动物学报.doc

技术方法 树鼩巴尔通体PCR方法的建立及初步应用 冯育芳, 邢 进, 付 瑞，巩 薇，贺争鸣，岳秉飞 （中国食品药品定研究院实验动物资源研究所） 摘要 目的方法结果（2.0×10-5 μg/mL），60份树鼩样本中有15份阳性样本，通过序列测定证实树鼩巴尔通体感染率为25%，与扩增结果完全相符，验证了该方法的可行性。结论 该方法的建立为树鼩巴尔通体的检测奠定了基础。 关键词；；【中图分类号】R33【文献标识码】A【文章编号】1671-7856（201）-0000-00 Doi:10.3969/j.issn.1671-7856.2014.006.000 Establishment and application of a PCR method for detection of Bartonella in tree shrew FENG Yu-fang, XING Jin, FU Rui, GONG Wei, HE Zheng-ming, YUE Bing-fei (Institute for?Laboratory Animal?Resources,?National Institutes for Food and Drug Control, Beijing 100050, China) [Abstract] Objective To establish an effective PCR assay for detection of Bartonell, and application of this assay in tree shrew. Methods Sequence of Bartonella was obtained from NCBI Genbank. Three pairs of primers were designed based on this sequence. One pair of primers was determined through amplifying the major strains in China. Sixty tree shrew blood samples were tested with this PCR assay. The positive amplified fragments were sequenced to verify the reliability of this method. Results A PCR method for detection of Bartonella is successfully established, with a high specificity and the sensitivity was of 2.0×10-5 μg/mL. Among the tested 60 blood samples, 15 positive cases were detected. Sequencing of the samples confirmed a 25% infection rate of Bartonella in the tree shrews, well consistent with amplification results, and verified the applicability of this detection method. Conclusion The establishment of this method provides the basis for detection of Bartonella in tree shrew. 【Key words】 Tree shrew; Bartonella, detection; PCR; Application [基金项目] 实验动物新品种的种群建立与质量标准化研究（2011BAI15B01）。 [作者冯育芳1981-），助理研究员，研究方向：实验动物微生物检测E-mail: fyf307@126.com。 [通讯作者(1960-)，男，研究员，研究方向：实验动物学。E-mail树鼩（tree shrew, Tupaia belangeri），树鼩科树鼩属的动物，许多分子和细胞结构近似于人类，对多种人类重要病毒易感，可作为重要人类病毒的动物模型[1]在肿瘤学、内分泌学、神经生物学、生殖生物学、免疫学等方面得到科研人员的肯定[2, 3]。而微生物背景检测是树鼩实验动物化的必备条件之一[2, 3]。 巴尔通体（Bartonella）是巴尔通体病的病原体能