二代测序建库中福尔马林固定石蜡包埋标本DNA超声打断方法的优化效果观察  

Optimization of DNA ultrasonic interruption method for Formalin-fixed and paraffin-embedding samples in the Second Generation Sequencing Database

作　　者：李余发[1] 许洁[1] LI Yufa;XU Jie(Pathology Department,Guangdong Provincial People’s Hospital of Southern Medical University(Guangdong Academy of Medical Sciences),Guangzhou,Guangdong 510010,China)

机构地区：[1]南方医科大学附属广东省人民医院<广东省医学科学院>病理科,广东广州510010

出　　处：《医药前沿》2023年第20期24-27,共4页Journal of Frontiers of Medicine

基　　金：广东省中医药局科研项目(20231008)。

摘　　要：目的:分析24例福尔马林固定石蜡包埋(FFPE)标本DNA采用新的改良的超声打断方法得到的结果与原打断方法得到结果的一致性,进一步优化目前福尔马林固定石蜡包埋样本DNA超声打断方法。方法:收集广东省人民医院病理科2023年4—5月进行二代测序检测病例24例,建库过程中根据使用的打断方法不同分为两组,1组采用原DNA打断方法(打断条件为:稀释后样本转移入配套的microTUBE AFA Fiber Snap-Cap打断管),2组采用改良DNA打断方法(打断条件为:稀释后样本转移入一次性超声波核酸打断耗材),比较两组实验结果是否一致。结果:两组实验结果得到的插入片段大小均>150bp,符合二代测序实验需要,差异无统计学意义(P>0.05)。结论:在临床实践中,二代测序建库过程中采用新的福尔马林固定石蜡包埋标本DNA超声打断方法可以达到原方法的实验效果,且更加经济实惠,从而能降低二代测序实验成本,值得在临床应用。Objective To investigate the consistency between the results of 24 Formalin-fixed and paraffin-embedding(FFPE)sample DNA using the new improved ultrasonic interruption method and the results obtained by the original interruption method,and further optimize the current formaldehyde fixed paraffin embedded sample DNA ultrasonic interruption method.Methods A total of 24 cases of second-generation sequencing were collected from the Department of Pathology of Guangdong Provincial People’s Hospital from April to May 2023,and were divided into two groups according to the different interruption methods used during the construction of the database.The first group was treated with the original DNA interruption method(the interruption condition was:After dilution,the samples were transferred into the matching microTUBE AFA Fiber Snap-Cap interrupting tube),and the improved DNA interrupting method was adopted in the second group(the interrupting condition was:After dilution,the samples were transferred into disposable ultrasonic nucleic acid interrupting consumables).The final experimental results of the two groups were compared.Results The insertion fragment sizes obtained from both experimental groups were greater than 150 bp,which meeted the needs of second-generation sequencing experiments,and the difference was not statistically significant(P>0.05).Conclusions In clinical practice,the new Formaldehyde fixed paraffin embedded DNA ultrasonic interruption method can achieve the experimental effect of the original method in the process of building the second generation sequencing database,and it is more economical,which can reduce the cost of the second generation sequencing experiment,it’s worth of the clinical promotion.

关 键 词：DNA 超声打断 福尔马林固定石蜡包埋 二代测序 

分 类 号：R445.1[医药卫生—影像医学与核医学]