Funato T, Satoh J, Sasaki T
Department of Clinical and Laboratory Medicine, Tohoku University, School of Medicine, Sendai.
Rinsho Byori. 1998 May;46(5):399-405.
We evaluated a real time quantitative PCR assay using dual-labeled fluorogenic probes for clinical application. Preliminary study using the house-keeping gene, beta-actin confirmed that this method was accurate and reproducible for the quantitative detection of the genes. The system also has merit with regard to the dynamic range of the starting target molecule determination. We then investigated DNA copies of cytomegalovirus (CMV) gene in vivo. The results demonstrated on association between the quantitation of CMV-DNA copies and clinical manifestation associated with CMV infection of immunodeficiency states or infantile hepatitis. It was also successful for quantitative estimation by RT-PCR. Namely, the assay made it possible to discriminate drug-sensitive leukemia cells from resistant cells based on the MDR1 gene and dCK gene. Real time quantitative PCR assay may be useful in a variety of clinical fields.
我们评估了一种使用双标记荧光探针的实时定量PCR检测方法用于临床应用。使用管家基因β-肌动蛋白的初步研究证实，该方法对于基因的定量检测准确且可重复。该系统在起始靶分子测定的动态范围方面也具有优势。然后我们研究了体内巨细胞病毒（CMV）基因的DNA拷贝数。结果表明，CMV-DNA拷贝数的定量与免疫缺陷状态或婴儿肝炎的CMV感染相关的临床表现之间存在关联。通过RT-PCR进行定量估计也很成功。也就是说，该检测方法能够基于多药耐药基因1（MDR1）和脱氧胞苷激酶（dCK）基因区分药物敏感白血病细胞和耐药细胞。实时定量PCR检测方法可能在各种临床领域中有用。
