Published online Mar 21, 2023. doi: 10.3748/wjg.v29.i11.1721
Peer-review started: October 14, 2022
First decision: January 3, 2023
Revised: January 13, 2023
Accepted: February 27, 2023
Article in press: February 27, 2023
Published online: March 21, 2023
Processing time: 153 Days and 23.8 Hours
Hepatitis B virus (HBV) genotype C infections has distinct clinical or virological traits including higher risk of hepatocellular carcinoma, lower response rate to interferon or prolonged hepatitis B e antigen-positive phase. As a likely answer to this issue, we have recently reported that the presence of two HBV Pol RT polymorphisms, rt269L and rt269I could contribute to unique traits of HBV genotype C.
For the identification between two rt269 types from chronic patients of genotype C2 endemic areas instead of time or labor consuming direct sequencing protocol, we sought to develop a novel simple and sensitive locked nucleotide probe based real-time polymerase chain reaction (LNA-RT-PCR) method capable of separating two rt269 types, rt269L type encoding leucine, ‘L’ (L1: CTC, L2: CTA) and rt269I type encoding isoleucine (ATC) from chronic hepatitis B (CHB) genotype C2 patients.
To develop a novel simple and sensitive LNA-RT-PCR method capable of identifying two rt269 types in CHB genotype C2 patients.
We designed appropriate primer and probe sets for LNA-RT-PCR for the separation of rt269 types. The developed LNA-RT-PCR method was applied to a total of 94 CHB patients of genotype C2 for the identification of two rt269 polymorphisms, and these results were compared with those obtained by a direct sequencing protocol.
The LNA-RT-PCR method could identify two rt269L and rt269I polymorphisms of three genotypes, two rt269L types [‘L1’ (WT) and ‘L2’] and one rt269I type (‘I’) in single (63 samples, 72.4%) or mixed forms (24 samples, 27.6%) in 87 (92.6% sensitivity) of 94 samples from Korean CHB patients.
The newly developed LNA-RT-PCR method could identify two rt269 polymorphisms, rt269L and rt269I, in CHB patients with genotype C2 infections. This method could be effectively used for the understanding of disease progression in genotype C2 endemic areas.
The newly developed LNA-RT-PCR method could identify three rt269 types, L1, L2 and I from CHB patients of genotype C2 with high-sensitivity and specificity. It could play a relevant role in the clinical management of CHB patients of genotype C2 infection.