Gastroenterology and hepatology from bed to bench · 2023
Development and evaluation of a semi-nested PCR technique for amplification and determination of different surface gene variation patterns in patients with chronic HBV infection: development of an amplification method for HBV S gene.
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Mokhtari Fedra, Kaboosi Hami, Mohebbi Seyed Reza, Asadzadeh Aghdaei Hamid, Zali Mohammad Reza
Аннотация
AIM: Development of an amplification method for further investigation of HBV S gene variation patterns.
BACKGROUND: Pre-S/S variants in patients with chronic HBV infection may contribute to the progression of liver damage and Hepatocellular carcinoma (HCC).
METHODS: This study was performed on ten patients with chronic HBV infection. Viral DNA was extracted from patient's plasma, primer design was performed, and a semi-nested PCR method was set up to amplify the pre-S/S region of HBV genome. Subsequently, sequencing was performed to analyze the variants of this region.
RESULTS: In the current study, the semi-nested PCR method was successfully set up, and types of variation in the studied samples were investigated.
CONCLUSION: Pre-S/S variants should be routinely determined in HBV carriers to help identify individuals who may be at a high risk of less favorable liver disease progression. This study showed that the technique could accurately amplify the pre-S/S region, and the product can be successfully used for variation detection by direct sequencing.
BACKGROUND: Pre-S/S variants in patients with chronic HBV infection may contribute to the progression of liver damage and Hepatocellular carcinoma (HCC).
METHODS: This study was performed on ten patients with chronic HBV infection. Viral DNA was extracted from patient's plasma, primer design was performed, and a semi-nested PCR method was set up to amplify the pre-S/S region of HBV genome. Subsequently, sequencing was performed to analyze the variants of this region.
RESULTS: In the current study, the semi-nested PCR method was successfully set up, and types of variation in the studied samples were investigated.
CONCLUSION: Pre-S/S variants should be routinely determined in HBV carriers to help identify individuals who may be at a high risk of less favorable liver disease progression. This study showed that the technique could accurately amplify the pre-S/S region, and the product can be successfully used for variation detection by direct sequencing.