用模糊综合评价法研究从外周血获取DNA标本的影响因素

目的：用模糊综合评价法研究从外周血获取DNA标本的主要影响因素，提供适合于分子流行病学调查时血液标本运输保存方式、血液标本类型、DNA提取方法。方法：采集40位健康志愿者静脉血，各自分成三种不同的类型(全血、血凝块、白细咆)，并随机将其分为四种方法进行保存，用四种常用方法提取DNA。使用模糊综合评价方法评价血液标本类型、DNA提取方法的优劣。结果：评价三种血液标本类型，血凝块提取DNA的综合评分最高，其次为全血和WBC；评价四种DNA提取方法，综合评分排列顺序为盐析法、TKM法、KI法、酚氯仿法。结论：各类样品应以低温运输与低温保存。如果开展较大规模的分子流行病学研究．综合各种因素，建议选取血凝块或全血样品，采用盐析法提取DNA。如特别要考虑DNA纯度可使用酚／氯仿法提取。

Studying Influential Factor of DNA Specimen Obtained from Blood Sample with Fussy Comprehension Evaluation

Objective To study the impact of different ways of transport and storage,DNA extraction methods,types of blood samples on the quantity and quality of isolated DNA in large-scale epidemiological studies,and apply them to the management of blood samples on-the-spot epidemiological studies.Methods Forty blood samples of health donors were divided into two group by random:A(R24hr),B(L24hr),C(R3m),D(L3m),each sample was made to three types(whole blood,clotted blood and WBC),from which DNA was extracted by four method of Phenol/chloroform, Salt,TKM and KI respectively. Analysis of ANOVA was used to compare the difference in quantity and purity of DNA extracted. Fussy comprehension evaluation was used to evaluate that which was the best of blood sample types and DNA extraction methods. Results When three types of blood samples were assessed,the composite score about clotted blood was the most,whole blood secondly and WBC thirdly. When DNA extraction methods were assessed,the composite score arranged in number order from big to small was Salt,TKM, KI, Phenol/chloroform method.Conclusion For extracting more yield and better purity of DNA from blood sample,all kinds sample should be keeped in low temperature. When molecular epidemiological studies were carried out,it is may be good to collect whole blood or clotted blood sample.About the extraction methods,yield of DNA extracted by Salt is the most;if quality of DNA was more considered,Phenol/chloroform will be recommended.