一种改良的人DNA微量快速检验技术

An improved method for fast detection of trace amount human DNA

目的探索一种以口腔粘膜脱落细胞为材料的安全、高效、低成本、敏感性高的人DNA微量快速检测方法。方法采用从漱口液或棉签擦拭口腔粘膜获取脱落细胞,应用混合树脂(Chelex100)煮沸沉淀法提取DNA;用PCR分别对线粒体特异性DNA片断和基因组中的特定基因进行扩增检测。结果通过对线粒体DNA中440 bp的非编码片段和染色体基因组中220 bp的乙醛脱氢酶DNA片段进行扩增,结果显示,从漱口液脱落细胞提取的DNA中可以稳定地扩增出上述两种DNA片断。结论建立了一种改良的人口腔粘膜脱落细胞DNA微量快速检验技术。该法取样方便,DNA样品获取量较大,一次取样可同时进行多项线粒体和基因组标志DNA片段的快速PCR检测。

Objectlve To explore and establish a safe,efficient,low cost and sensitive fast detection method of trace amount human DNA from easily accessible materials.Methods A simple Chelex100 boiling-precipitation method was developed to extract clean DNA from oral exfoliated cells. The obtained DNA sample was used as template to PCR-amplify specific DNA fragments from mitochondrial and genomlc DNA. Results A 440 bp fragment from the noncoding region of mitochondrial DNA and a 220 bp ALDH gene fragment from genomic DNA can be efficiently and readily amplified from DNA samples derived from oral exfoliated cells. Conclusion An improved method for fast detection of trace amount human DNA was established. The method can easily obtain clean DNA sampies from gargles that can be used for fast multiple detections of any mitochondrial and genomic marker DNA fragments by PCR.