自动化工作站与Chelex-100法对血样本检验效果比较

Comparison of the DNA extraction effect of blood samples by using automation workstation and Chelex-100 method

目的 比较自动化工作站与Chelex-100法对不同浓度血液样本的检验效果.方法 取新鲜血液,按倍量稀释法制成2～1 024倍10种不同浓度的血样本,分别采用自动化工作站和Chelex-100法提取模板DNA,应用IdentifilerPlus试剂盒进行扩增及毛细管电泳检测,并对电泳结果进行比较.结果 稀释2～32倍的血样本,采用两种方法的检验成功率均可达到100％;稀释64倍的样本,Chelex-100法成功率为41.3％、完整分型数为78.3％,自动化工作站成功率为13.0％、完整分型数为43.5％,两者比较,差异均有统计学意义（P＜0.01）;稀释128倍的样本,成功率在Chelex-100法（8.7％）和自动化工作站（2.4％）之间比较,其差异无统计学意义（P＞0.05）;稀释256倍以上,两种方法均无法获得分型结果.结论 自动化工作站适用于一定浓度血样的检验,而对于微量血样,Chelex-100法的效果可能更优.

Objective Comparison of the DNA extraction effect of different concentration blood samples by using automation workstation and Chelex-100 methods. Methods Fresh blood was diluted 2 - 1 024 times according to double dilution method. Next, automation workstation and Chelex-100 methods were used to extract DNA template from those 10 different concentrations of blood samples, respectively. Then the template DNA were performed PCR with Identifiler Plus kit and capillary eleetrophoresis, the electrophoresis results were compared. Results The test success rates of the diluted 2 -32 times blood samples test with both methods were 100% ;The diluted 64 times blood samples, the test success rate and complete typing number rate of Chelex-100 method was 41.3% and 78.3% , the test success rate and complete typing number rate of automation workstation method was 13.0% and 43.5% , there were significant difference between both methods（P 〈0. 01 ） ; the diluted 128 times blood samples, the test success rate of Chelex-100 and automation workstation methods was 8.7% and 2. 4% , respectively, there was no significant difference between both methods （ P 〉 0. 05 ） ; when the blood samples were diluted 256 times and more, the STR typing were failed with both methods. Conclusion Automation workstation method was suitable for DNA extraction of a certain concentration blood samples. Chelex-100 method may be better for the trace blood samples.