血液和骨髓标本中常见PCR反应抑制物的探究与分析  

Exploration and Analysis of PCR-inhibitory Components in Peripheral Blood and Bone Marrow Samples——Review

作　　者：沈克锋[1] 杨默[1] 江千里[1] 

机构地区：[1]南方医科大学南方医院血液科,广东广州510515

出　　处：《中国实验血液学杂志》2014年第3期842-846,共5页Journal of Experimental Hematology

摘　　要：PCR反应具有特异性强、灵敏度高、方便快捷、高效安全及对标本要求低的特点,是检测标本中DNA或mRNA的首选方法,大量应用于临床和科研。近年来逆转录荧光定量PCR技术(real-time reverse transcription PCR,qRT-PCR)取得了飞速发展,检测灵敏度不断提高,有的甚至可以检测低于10个拷贝数的基因。技术的进步必然会对标本提出更高的要求,因此有必要重新审视血液和骨髓液中可能存在的PCR反应抑制物。本文总结了血液和骨髓液中常见的PCR反应抑制物及其可能存在的抑制机制和解决措施,重点阐述了抗凝剂肝素的使用对于PCR反应的影响。本文可在一定程度上指导对PCR(特别是qRT-PCR)检测结果的评估,如对于某些低表达基因定量检测结果临床意义的判断,或者排除标本中PCR抑制成分所导致的假阴性等。PCR is an enabling and preferred technology for the detection of DNA or mRNA in genomic age, and has been widely used in molecular medicine, biotechnology, microbiology and diagnostics. PCR has many excellent traits, such as high specificity and sensitivity, short handle time, reliable and effective results, low demand for sample. In recent years, real-time reverse transcription PCR （qRT-PCR） has acquired an impressive improvement in the detection of mRNA, it has a perfect sensitivity and can quantify some rare mRNA copies as low as less ten. The progress in PCR results in a more strict demand for sample, so it is necessary to take the potential PCR-inhibitory components in peripheral blood and bone marrow samples into account again. This review explores the potential PCR-inhibitory components in peripheral blood and bone marrow samples, and discusses the possible mechanism and solutions for the inhibition effect, also emphasizes the potent effect of the frequently-used anticoagulant heparin sodium in PCR. This article will be helpful to assess the PCR detection results （especially qRT-PCR ） in some degree, for example, can use it to decide whether the PCR detection results are reasonable for some low-expression genes, or use it to eliminate the false negative results generated by PCR-inhibitory components in sample.

关 键 词：血液 骨髓标本 qRT—PCR PCR抑制物 

分 类 号：R446.1[医药卫生—诊断学]