摘要
目的研发生物兼容性好并能模拟实际临床样本性质的基因检测质控品。方法质控品研究。通过将含有特定基因突变的细胞或DNA包埋于γ-聚谷氨酸与明胶交联形成的生物基质,再经过福尔马林固定及石蜡包埋制备质控品。采用傅立叶红外光谱及扫描电镜分析质控品的形态结构。通过对质控品不同部位的DNA提取量、突变频率进行检测验证其重复性和均一性。将质控品放置于25℃条件下保存16周,通过检测DNA提取量、突变频率以及DNA片段分布验证其稳定性。结果傅立叶红外光谱及扫描电镜分析结果表明本研究制备的质控品通过交联作用形成,具有致密的三维骨架结构。质控品不同部位的DNA提取量及突变频率波动较小(CV<10%)。质控品在25℃条件下保存16周DNA提取量及突变频率波动较小(CV<5%),并且DNA片段分布范围与质控品制备当天比较没有明显变化。结论本研究成功制备了基于交联生物基质的基因检测质控品,具有良好的重复性、均一性及稳定性。
ObjectiveTo develop genetic testing quality control material which has good biological compatibility that can simulate the real sample properties.MethodsStudy of the quality control material.The quality control matertial was prepared by embedding the cells and DNA containing specific gene mutations in a biological matrix formed by cross-linkingγ-polyglutamic acid and gelatin,followed by formalin fixation and paraffin embedding.Fourier infrared spectroscopy and scanning electron microscopy were used to analyze the morphology and structure of the quality control material.The repeatability and homogeneity were verified by detecting the DNA extraction amount and mutation frequency at different parts of the quality control material.The quality control material was stored at 25℃ for 16 weeks,and the stability was verified by detecting the DNA extraction amount,mutation frequency and DNA fragment distribution.ResultsThe results of Fourier infrared spectroscopy and scanning electron microscopy analysis indicated that the quality control material prepared in this study was formed by cross-linking and had a dense three-dimensional framework structure.The amount of DNA extraction and mutation frequency from different parts of the quality control material varied within a small range(CV<10%).After storage at 25℃for 16 weeks,the DNA extraction amount and mutation frequency of the quality control material varied within a small range(CV<5%),and the distribution range of the DNA fragments did not show significant change compared to the day of preparation of the quality control material.ConclusionThis study successfully prepared a genetic testing quality control material based on cross-linked biological matrix,which exhibited good repeatability,homogeneity and stability.
作者
谢建飞
李春妞
马颖颖
Xie Jianfei;Li Chunniu;Ma Yingying(Shanghai Greenikon Clinical Laboratory,Shanghai 201702,China)
出处
《中华检验医学杂志》
北大核心
2025年第8期1085-1090,共6页
Chinese Journal of Laboratory Medicine
关键词
基因检测
质控品
Γ-聚谷氨酸
明胶
石蜡包埋
Genetic testing
Quality control material
γ-polyglutamic acid
Gelatin
Paraffin embedding
