博尔纳病病毒荧光定量pcr 试剂盒pcr 扩增缓冲液的选择和优化.docVIP

博尔纳病病毒荧光定量PCR试剂盒PCR扩增缓冲液的选择和优化 金戈1,2,3，徐晓艳1,2,3，张亮1,2,3，马丽华1,2,3，黄荣忠1,2,3，邓婧1,2,3，李文娟1,2,3，房亮2,3，谢鹏1,2,3（重庆医科大学附属第一医院神经内科1；重庆医科大学神经科学研究中心2，重庆市神经生物学重点实验室3，重庆 400016） [摘要] 目的 选择并优化适合博尔纳病病毒实时荧光定量PCR诊断试剂盒反应体系中的扩增缓冲液，使PCR反应体系对该病毒p24模板基因的扩增效率得到明显提高。方法方法太简单，写详细一点，写明具体比较的酸、添加剂、以及不同的浓度等具体的方法步骤 通过常规PCR比较选择三3种常用酸（盐酸、磷酸、硫酸）滴定的缓冲液体系，并在其基础上进一步摸索适当的添加剂进行优化组合，以得到最佳的扩增效果，最后用荧光定量PCR进行对比验证。结果 结果太简单，应该写明各种优化条件的具体参数，比如各种添加剂的组合及浓度等在滴定酸上选择了效果较好的硫酸，在添加剂的优化组合上选择效果最好的TBTT buffer，在其与Takara PCR buffer的对比中，TBTT buffer效果好于后者。结论 成功研制出适合扩增博尔纳病病毒p24模板基因的扩增缓冲液，为开发博尔纳病病毒实时荧光定量PCR诊断试剂盒奠定了基础。 方法太简单，写详细一点，写明具体比较的酸、添加剂、以及不同的浓度等具体的方法步骤 结果太简单，应该写明各种优化条件的具体参数，比如各种添加剂的组合及浓度等 [关键词] 关键词太长，可改为：PCR扩增缓冲液；PCR试剂盒；博尔纳病病毒博尔纳病病毒荧光定量PCR试剂盒；扩增缓冲液；优化 关键词太长，可改为：PCR扩增缓冲液；PCR试剂盒；博尔纳病病毒 [中图法分类号] R373.9; R185 [文献标识码] A Selection and optimization of PCR buffer in the development of Borna Disease Virus Real-time PCR diagnostic kit Jin Ge1,2,3,Xu Xiaoyan1,2,3,Zhang Liang1,2,3,Ma Lihua1,2,3,Huang Rongzhong1,2,3,Deng Jing1,2,3,Li Wenjuan1,2,3,Fang Liang2,3,Xie Peng1,2,3 (Department of Neurology1, First Affiliated Hospital; Neuroscience Center2; Key Laboratory of Neurobiology Designated by the Municipality of Chongqing3, Chongqing Medical University, Chongqing 400016, China) [Abstract] Objective Select and optimize the PCR buffer for Borna disease virus Real-time fluorescence quantitative PCR diagnostic kit to significantly improve the PCR amplification efficiency to BDV p24 gene. Methods Conventional PCR was used to contrast the PCR buffers which titrated with three commmmol/Lon acids respectively,and further explored the appropriate PCR additive mix to enhanced the quantitative and qualitative output of PCR and finally verified the efficiency of some PCR buffers by Real-time PCR. Results Compared to other acids,sulfuric acid is more suitable to titrate the buffer. Furthermore, Real-time PCR results indicated that TBTT buffer is the most effective buffer compare to other PCR additive mix and is more effective to ampli