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Updated: Jul 17, 2026
Real-Time Polymerase Chain Reaction-Based Detection and Quantification of Hepatitis B Virus DNA
Published on: December 15, 2023
Analysis of differential gene expression between chronic hepatitis B patients and asymptomatic hepatitis B carriers
Cuilan Tang1, Zhi Chen, Guoping Peng
1Key Laboratory of Health Ministry, Institute of Infectious Diseases, First Affiliated Hospital, School of Medicine, Zhejiang University, Zhejiang, China.
Insights
Researchers identified differentially expressed genes in individuals with chronic hepatitis B (CHB) and asymptomatic HBV carriage (ASC). These genes offer insights into distinct HBV infection states and potential therapeutic targets.
Area of Science:
- * Hepatology and Virology
- * Immunology and Molecular Biology
Background:
- * Chronic hepatitis B virus (HBV) infection is a major global health concern.
- * Asymptomatic HBV carriage (ASC) and chronic hepatitis B (CHB) represent distinct immunological states with different prognoses.
Purpose of the Study:
- * To identify differentially expressed genes between CHB and ASC states.
- * To elucidate molecular mechanisms underlying these distinct HBV infection states.
Main Methods:
- * Suppression subtractive hybridization (SSH) was employed to create subtracted cDNA libraries.
- * Dot blot hybridization and quantitative real-time PCR (qPCR) were used for gene screening and validation.
Main Results:
- * SSH identified subtracted cDNA clones from peripheral blood mononuclear cells of CHB and ASC subjects.
- * Dot blot hybridization screened 88 clones, identifying 29 differentially expressed genes.
- * Real-time PCR confirmed differential expression of three genes in 23 CHB and 21 ASC subjects.
Conclusions:
- * Differentially expressed genes, including novel ones, were isolated between CHB and ASC states.
- * Up-regulated genes in CHB are associated with inflammatory responses.
- * Identified genes may provide insights into HBV infection mechanisms and therapeutic strategies.
Background And Aims:
Chronic hepatitis B virus (HBV) infection remains a serious global health problem, inducing a spectrum of diseases, including asymptomatic HBV carriage (ASC) and chronic hepatitis B (CHB). ASC and CHB represent different immunological states and their prognoses are diverse. To clarify molecular mechanisms underlying the two infection states, the differentially expressed genes between the two states were screened and identified.
Methods:
Subtracted complementary DNA libraries by suppression subtractive hybridization, dot blot hybridization and quantitative real-time PCR were used to identify the differentially expressed genes between subjects with CHB and those with ASC.
Results:
RNA from peripheral blood mononuclear cells from CHB and ASC subjects was subjected to suppression subtractive hybridization and resulted in isolation of subtracted complementary DNA clones. Eighty-eight randomly sampled clones were rescreened by dot blot hybridization, from which 29 clones were identified as differentially expressed genes. The differential expression of three genes was confirmed by real-time PCR in 23 subjects with CHB and 21 with ASC.
Conclusions:
Differentially expressed genes in peripheral blood mononuclear cells between CHB and ASC have been isolated by suppression subtractive hybridization, including some new genes. Of the up-regulated genes in CHB, most are known to be responsive to inflammatory conditions. These genes might provide clues in elucidating the mechanisms of the two different HBV infection states and designing therapeutic targets for HBV infection.
