Locked nucleic acid real-time polymerase chain reaction method identifying two polymorphisms of hepatitis B virus genotype C2 infections, rt269L and rt269I

Figure 3 Multiprobe locked nucleic acid real-time polymerase chain reaction for discrimination among three types of polymorphisms in the rt269 codon. Amplification curves are shown on the left, and melting peaks are shown on the right. A: With L1 wild-type DNA templates, L1-type specific signals in the FAM channel (solid) were detected, showing their dominant amplification and distinct melting temperatures (T_m), with minimal cross signals of amplification and melting peaks generated by weak cross hybridizations of the other probes (I and L2), which were differentiated from the T_m values for I and L2 detection; B: For I variant-type DNA templates, I-type specific signals in the Hex channel (dotted) were detected, showing their exclusive amplifications and distinct T_m values, with no cross signals; C: For L2 variant-type DNA templates, amplification curves showed weak cross signals, but melting peaks were distinct with no cross signals.