Selective detection of HBV pre-genomic RNA in chronic hepatitis B patients using a novel RT-PCR assay
In chronic hepatitis B (CHB) patients, quantification of HBV pgRNA in plasma has the potential to provide information on disease prognosis and liver injury or histopathology. However, current methods for detecting HBV pgRNA present technical difficulties due to the co-existence of HBV DNA in plasma...
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| Veröffentlicht in: | Clinical and experimental medicine 2023-12, Vol.23 (8), p.5281-5289 |
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| creator | Nguyen, Ung Dinh Le Do, Quyen Vu, Quynh Anh Nguyen Trieu, Nguyet Thi Dao, Trang Thuy Van Le, Nam Nguyen, Son Thai Hoang, Tuyen Tien Nguyen, Chinh Trong Nguyen, Thang Hong Van Nguyen, Dien Ho, Tho Huu |
| description | In chronic hepatitis B (CHB) patients, quantification of HBV pgRNA in plasma has the potential to provide information on disease prognosis and liver injury or histopathology. However, current methods for detecting HBV pgRNA present technical difficulties due to the co-existence of HBV DNA in plasma samples. We have successfully established a novel one-step RT-PCR assay that allows selective quantification of HBV pgRNA. Two cohorts of participants were recruited for assay validation, including treatment-naïve patients with CHB and HBeAg-positive CHB patients who were treated with Tenofovir and monitored for 6 months to assess the predictive value of baseline HBV RNA for HBeAg seroclearance. Statistical analysis was performed using MedCalc version 20.019 software. The novel selective one-step RT-PCR assay for detecting HBV pgRNA was validated with a limit of detection of 100 copies/mL. The assay was able to selectively measure HBV pgRNA even in the presence of excess HBV rcDNA. In treatment-naïve CHB patients, HBV pgRNA levels were significantly lower than HBV DNA concentration. Serum HBV DNA levels and HBeAg status were positively associated with HBV pgRNA. Baseline serum HBV pgRNA levels were found to be strong predictors of HBeAg seroclearance after 6 months of Tenofovir treatment. The study presents a novel RT-PCR assay that allows accurate measurement of plasma HBV pgRNA in chronic hepatitis B patients, even in the presence of excess HBV DNA. The assay is highly selective and represents a significant advancement with potential for further breakthroughs in understanding the clinical significance of HBV pgRNA.
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| doi_str_mv | 10.1007/s10238-023-01162-6 |
| format | Article |
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Graphical abstract</description><subject>Antiviral Agents</subject><subject>Deoxyribonucleic acid</subject><subject>DNA</subject><subject>DNA, Viral - genetics</subject><subject>Genomics</subject><subject>Hematology</subject><subject>Hepatitis B</subject><subject>Hepatitis B e antigen</subject><subject>Hepatitis B e Antigens</subject><subject>Hepatitis B virus - genetics</subject><subject>Hepatitis B, Chronic - diagnosis</subject><subject>Hepatitis B, Chronic - drug therapy</subject><subject>Humans</subject><subject>Internal Medicine</subject><subject>Liver diseases</subject><subject>Medicine</subject><subject>Medicine & Public Health</subject><subject>Oncology</subject><subject>Patients</subject><subject>Polymerase chain reaction</subject><subject>Reverse Transcriptase Polymerase Chain Reaction</subject><subject>RNA</subject><subject>Statistical analysis</subject><subject>Tenofovir</subject><subject>Tenofovir - therapeutic use</subject><issn>1591-9528</issn><issn>1591-8890</issn><issn>1591-9528</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2023</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNp9kU1PGzEQhi3UClLoH-BQWeLCxe34az-OJIKCFBUU0l4tZ3c2WbSxU3s3Ev8eh6Sl4tDL-LX8zOvRvIScc_jKAfJvkYOQBUuFAeeZYNkRGXFdclZqUXz4R5-QTzE-AXBdSDgmJzLXueBajQg-YodV326R1tjvlHfUN_R2_ItuArIlOr9uKzr7cUVbR6tV8C5dV7ixfdu3kY7pTqHrIx1i65bUUue32NHZnD1MZtTGaJ_PyMfGdhE_H85T8vPmej65ZdP773eTqymrpMh6ZheFzksLUulMFILnjbK1zqq6VrXlClSN2UKhkkWmuK2UzpsCAaTkDWitS3lKLve-m-B_Dxh7s25jhV1nHfohGlFokJDcd-jFO_TJD8Gl6YwoQeQAWkGixJ6qgo8xYGM2oV3b8Gw4mF0IZh-CScW8hmCy1PTlYD0s1lj_bfmz9QTIPRDTk1tiePv7P7YvhfyPRQ</recordid><startdate>20231201</startdate><enddate>20231201</enddate><creator>Nguyen, Ung Dinh</creator><creator>Le Do, Quyen</creator><creator>Vu, Quynh Anh Nguyen</creator><creator>Trieu, Nguyet Thi</creator><creator>Dao, Trang Thuy</creator><creator>Van Le, Nam</creator><creator>Nguyen, Son Thai</creator><creator>Hoang, Tuyen Tien</creator><creator>Nguyen, Chinh Trong</creator><creator>Nguyen, Thang Hong</creator><creator>Van Nguyen, Dien</creator><creator>Ho, Tho Huu</creator><general>Springer International Publishing</general><general>Springer Nature B.V</general><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7T5</scope><scope>7TK</scope><scope>H94</scope><scope>K9.</scope><scope>7X8</scope><orcidid>https://orcid.org/0000-0002-8363-1427</orcidid></search><sort><creationdate>20231201</creationdate><title>Selective detection of HBV pre-genomic RNA in chronic hepatitis B patients using a novel RT-PCR assay</title><author>Nguyen, Ung Dinh ; Le Do, Quyen ; Vu, Quynh Anh Nguyen ; Trieu, Nguyet Thi ; Dao, Trang Thuy ; Van Le, Nam ; Nguyen, Son Thai ; Hoang, Tuyen Tien ; Nguyen, Chinh Trong ; Nguyen, Thang Hong ; Van Nguyen, Dien ; Ho, Tho Huu</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c326t-ab8579a0345628217f4ad56cdd4da1404de6b4e438641ac457f8e00331f055593</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2023</creationdate><topic>Antiviral Agents</topic><topic>Deoxyribonucleic acid</topic><topic>DNA</topic><topic>DNA, Viral - genetics</topic><topic>Genomics</topic><topic>Hematology</topic><topic>Hepatitis B</topic><topic>Hepatitis B e antigen</topic><topic>Hepatitis B e Antigens</topic><topic>Hepatitis B virus - genetics</topic><topic>Hepatitis B, Chronic - diagnosis</topic><topic>Hepatitis B, Chronic - drug therapy</topic><topic>Humans</topic><topic>Internal Medicine</topic><topic>Liver diseases</topic><topic>Medicine</topic><topic>Medicine & Public Health</topic><topic>Oncology</topic><topic>Patients</topic><topic>Polymerase chain reaction</topic><topic>Reverse Transcriptase Polymerase Chain Reaction</topic><topic>RNA</topic><topic>Statistical analysis</topic><topic>Tenofovir</topic><topic>Tenofovir - therapeutic use</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Nguyen, Ung Dinh</creatorcontrib><creatorcontrib>Le Do, Quyen</creatorcontrib><creatorcontrib>Vu, Quynh Anh Nguyen</creatorcontrib><creatorcontrib>Trieu, Nguyet Thi</creatorcontrib><creatorcontrib>Dao, Trang Thuy</creatorcontrib><creatorcontrib>Van Le, Nam</creatorcontrib><creatorcontrib>Nguyen, Son Thai</creatorcontrib><creatorcontrib>Hoang, Tuyen Tien</creatorcontrib><creatorcontrib>Nguyen, Chinh Trong</creatorcontrib><creatorcontrib>Nguyen, Thang Hong</creatorcontrib><creatorcontrib>Van Nguyen, Dien</creatorcontrib><creatorcontrib>Ho, Tho Huu</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Immunology Abstracts</collection><collection>Neurosciences Abstracts</collection><collection>AIDS and Cancer Research Abstracts</collection><collection>ProQuest Health & Medical Complete (Alumni)</collection><collection>MEDLINE - Academic</collection><jtitle>Clinical and experimental medicine</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Nguyen, Ung Dinh</au><au>Le Do, Quyen</au><au>Vu, Quynh Anh Nguyen</au><au>Trieu, Nguyet Thi</au><au>Dao, Trang Thuy</au><au>Van Le, Nam</au><au>Nguyen, Son Thai</au><au>Hoang, Tuyen Tien</au><au>Nguyen, Chinh Trong</au><au>Nguyen, Thang Hong</au><au>Van Nguyen, Dien</au><au>Ho, Tho Huu</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Selective detection of HBV pre-genomic RNA in chronic hepatitis B patients using a novel RT-PCR assay</atitle><jtitle>Clinical and experimental medicine</jtitle><stitle>Clin Exp Med</stitle><addtitle>Clin Exp Med</addtitle><date>2023-12-01</date><risdate>2023</risdate><volume>23</volume><issue>8</issue><spage>5281</spage><epage>5289</epage><pages>5281-5289</pages><issn>1591-9528</issn><issn>1591-8890</issn><eissn>1591-9528</eissn><abstract>In chronic hepatitis B (CHB) patients, quantification of HBV pgRNA in plasma has the potential to provide information on disease prognosis and liver injury or histopathology. However, current methods for detecting HBV pgRNA present technical difficulties due to the co-existence of HBV DNA in plasma samples. We have successfully established a novel one-step RT-PCR assay that allows selective quantification of HBV pgRNA. Two cohorts of participants were recruited for assay validation, including treatment-naïve patients with CHB and HBeAg-positive CHB patients who were treated with Tenofovir and monitored for 6 months to assess the predictive value of baseline HBV RNA for HBeAg seroclearance. Statistical analysis was performed using MedCalc version 20.019 software. The novel selective one-step RT-PCR assay for detecting HBV pgRNA was validated with a limit of detection of 100 copies/mL. The assay was able to selectively measure HBV pgRNA even in the presence of excess HBV rcDNA. In treatment-naïve CHB patients, HBV pgRNA levels were significantly lower than HBV DNA concentration. Serum HBV DNA levels and HBeAg status were positively associated with HBV pgRNA. Baseline serum HBV pgRNA levels were found to be strong predictors of HBeAg seroclearance after 6 months of Tenofovir treatment. The study presents a novel RT-PCR assay that allows accurate measurement of plasma HBV pgRNA in chronic hepatitis B patients, even in the presence of excess HBV DNA. The assay is highly selective and represents a significant advancement with potential for further breakthroughs in understanding the clinical significance of HBV pgRNA.
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| subjects | Antiviral Agents Deoxyribonucleic acid DNA DNA, Viral - genetics Genomics Hematology Hepatitis B Hepatitis B e antigen Hepatitis B e Antigens Hepatitis B virus - genetics Hepatitis B, Chronic - diagnosis Hepatitis B, Chronic - drug therapy Humans Internal Medicine Liver diseases Medicine Medicine & Public Health Oncology Patients Polymerase chain reaction Reverse Transcriptase Polymerase Chain Reaction RNA Statistical analysis Tenofovir Tenofovir - therapeutic use |
| title | Selective detection of HBV pre-genomic RNA in chronic hepatitis B patients using a novel RT-PCR assay |
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