Microfluidic leukocyte isolation for gene expression analysis in critically ill hospitalized patients
Microfluidic leukocyte isolation for gene expression analysis in critically ill hospitalized patients
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DOI：
10.1373/clinchem.2007.099150
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发表时间：
2008-05-01
影响因子：
9.3
通讯作者：
Toner, Mehmet
中科院分区：
文献类型：
--
作者：
Russom, Aman;Sethu, Palaniappan;Toner, Mehmet
背景：微阵列技术正在成为诊断、治疗和预后应用的有力工具。目前，关于从血液白细胞群中分离核酸用于后续表达分析的最佳技术尚无共识。目前的收集和处理技术在临床环境中提出了重大挑战。在这里，我们报告了一种新的微流控白细胞核酸分离技术的临床验证，该技术可用于危重病人的基因表达分析，住院患者可以很容易地用于小容量血液。方法：对烧伤和严重钝性创伤住院患者的全血进行微流控和标准大尺度白细胞分离处理。通过RNA数量、质量和全基因组表达模式的并排比较，临床验证了微流控技术。结果：采用微流控处理方案，从0.5 mL全血中获得足够量的总RNA进行全基因组表达分析。我们发现使用两种方法处理的样本的白细胞表达模式是一致的，并且由于收获方法而引入的变异性比个体之间存在的变异性要少。结论：新型微流控分离方法可实现白细胞分离
BACKGROUND: Microarray technology is becoming a powerful tool for diagnostic, therapeutic, and prognostic applications. There is at present no consensus regarding the optimal technique to isolate nucleic acids from blood leukocyte populations for subsequent expression analyses. Current collection and processing techniques pose significant challenges in the clinical setting. Here, we report the clinical validation of a novel microfluidic leukocyte nucleic acid isolation technique for gene expression analysis from critically ill, hospitalized patients that can be readily used on small volumes of blood.METHODS: We processed whole blood from hospitalized patients after burn injury and severe blunt trauma according to the microfluidic and standard macroscale leukocyte isolation protocol. Side-by-side comparison of RNA quantity, quality, and genome-wide expression patterns was used to clinically validate the microfluidic technique.RESULTS: When the microfluidic protocol was used for processing, sufficient amounts of total RNA were obtained for genome-wide expression analysis from 0.5 mL whole blood. We found that the leukocyte expression patterns from samples processed using the 2 protocols were concordant, and there was less variability introduced as a result of harvesting method than there existed between individuals.CONCLUSIONS: The novel microfluidic approach achieves leukocyte isolation in