Abstract
We demonstrated the application of IM-PCR, intercalation monitoring PCR, to quantify HCV RNA of serum samples from patients with chronic hepatitis C by performing PCR in the presence of oxazole yellow derivative, a fluorescent DNA intercalative dye, and monitoring the fluorescence intensity of the PCR reaction mixture in the course of PCR cycles. The assay gave the efficient and reproducible results in clinically useful dynamic range bellow 10-6 copies of HCV RNA for interferon therapy. We also reported here our novel fluorescent DNA probe, oxazole yellow (YO)-linked oligonucleotide complementary to a target DNA/RNA, which can enhance the fluorescence on hybridizing with a target nucleotide and its applicability to construct an assay of a sequence specific homogeneous detection of HCV RNA in clinical samples in conjunction with RT-PCR.
