Protocol for the use of self-reporting duplex mutation primers to detect PCR products in the diagnosis of HBV.
Quantitative measurements of serum hepatitis B virus (HBV) DNA are useful for tailoring of treatment schedules and the monitoring of HBV replication during therapy. We developed a novel fluorescence-based quantitative real-time PCR for quantitating HBV DNA based on the duplex mutation primers principle, in which signal is generated by melting a duplex mutation primer during renaturation. The duplex mutation primers are much more specific than double-stranded DNA dyes like SYBR Green I and, unlike other probes, do not require the double-labeled synthesis of fluorophore and quencher on the same molecule.
The Faculty of Laboratory Medicine and Tropical Medicine, Hainan Medical College, HaiKou, People's Republic of China. xiaqianfeng@sina.com
Qian-Feng
QF
Xia
D020137:Base Pair Mismatch; D052160:Benzothiazoles; D017931:DNA Primers / Q000235:genetics*; D004279:DNA, Viral / Q000032:analysis / Q000097:blood / Q000235:genetics* / Q000302:isolation & purification; D003959:Diamines; D006515:Hepatitis B virus / Q000235:genetics* / Q000302:isolation & purification*; D006801:Humans; D025202:Molecular Diagnostic Techniques / Q000379:methods*; D009154:Mutation; D009838:Oligodeoxyribonucleotides / Q000235:genetics / Q000378:metabolism; D009930:Organic Chemicals / Q000378:metabolism; D011804:Quinolines; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D013050:Spectrometry, Fluorescence
D052160:Benzothiazoles; D017931:DNA Primers; D004279:DNA, Viral; D003959:Diamines; D009838:Oligodeoxyribonucleotides; D009930:Organic Chemicals; D011804:Quinolines; C098022:SYBR Green I