Detection of lamivudine and adefovir resistance mutations among Filipino patients with chronic hepatitis B using line probe assay

Despite the effectiveness of nucleoside/nucleotide analogues in the treatment of chronic hepatitis B (CHB), their long-term administration is associated with the emergence of resistant hepatitis B virus (HBV) mutants. In this study, mutations resulting in antiviral resistance in HBV DNA samples isolated from 23 CHB patients (nine treatment naïve and 14 treated previously) were studied using a line probe assay (INNO-LiPA HBV DR; Innogenetics) and ultradeep pyrosequencing (UDPS) methods. Whilst the INNO-LiPA HBV DR showed no resistance mutations in HBV DNA samples from treatment-naive patients, mutations mediating lamivudine resistance were detected in three samples by UDPS. Among patients who were treated previously, 19 mutations were detected in eight samples using the INNO-LiPA HBV DR and 29 mutations were detected in 12 samples using UDPS. All mutations detected by the INNO-LiPA HBV DR were also detected by UDPS. There were no mutations that could be detected by INNO-LiPA HBV DR but not by UDPS. A total of ten mutations were detected by UDPS but not by INNO-LiPA HBV DR, and the mean frequency of these mutations was 14.7 %. It was concluded that, although INNO-LiPA HBV DR is a sensitive and practical method commonly used for the detection of resistance mutations in HBV infection, UDPS may significantly increase the detection rate of genotypic resistance in HBV at an early stage.

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Evaluation of a line probe assay for identification of hepatitis B virus precore variants in serum from chronic hepatitis B carriers

Journal of Virological Methods ◽

10.1016/j.jviromet.2003.09.014 ◽

2003 ◽

Vol 114 (1) ◽

pp. 97-103 ◽

Cited By ~ 2

Author(s):

Charlotte L. Cameron-Wilson ◽

Peter Muir ◽

Anna L. Ballard ◽

Sally Corden ◽

Elizabeth H. Boxall ◽

...

Keyword(s):

Chronic Hepatitis ◽

Hepatitis B Virus ◽

Hepatitis B ◽

Chronic Hepatitis B ◽

Line Probe Assay ◽

B Virus ◽

Probe Assay

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Comparison of Sequence Analysis and INNO-LiPA HBV DR Line Probe Assay in Patients with Chronic Hepatitis B

Journal of Chemotherapy ◽

10.1179/joc.2005.17.5.514 ◽

2005 ◽

Vol 17 (5) ◽

pp. 514-520 ◽

Cited By ~ 17

Author(s):

R.Y. Sertoz ◽

S. Erensoy ◽

S. Pas ◽

U.S. Akarca ◽

F. Ozgenc ◽

...

Keyword(s):

Chronic Hepatitis ◽

Sequence Analysis ◽

Hepatitis B ◽

Chronic Hepatitis B ◽

Line Probe Assay ◽

Probe Assay

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COLD-PCR Method for Early Detection of Antiviral Drug-Resistance Mutations in Treatment-Naive Children with Chronic Hepatitis B

Diagnostics ◽

10.3390/diagnostics10070491 ◽

2020 ◽

Vol 10 (7) ◽

pp. 491

Author(s):

Thuy Thi Bich Phung ◽

Son Van Chu ◽

Son Thien Vu ◽

Hanh Thi Pham ◽

Hang Minh Nguyen ◽

...

Keyword(s):

Chronic Hepatitis ◽

Hepatitis B ◽

Chronic Hepatitis B ◽

Single Point ◽

Antiviral Drug ◽

Multiple Point ◽

Resistance Mutations ◽

Hbv Genotypes ◽

Treatment Naïve ◽

Pcr Method

We investigated Nucleos(t)ide-analogue (NA)-resistance mutations (mt) in 142 treatment-naive children with Chronic Hepatitis B (CHB), using a sensitive co-amplification at lower denaturation temperature (COLD)-PCR with Sanger DNA sequencing. An NA resistance-associated mt in the hepatitis B virus (HBV) reverse transcriptase (RT) was found in 66.2% of the patients, with nonclassical mt contributing the most (64.8%). Significantly higher frequencies of Lamivudine (LMV) and Adefovir dipivoxil (ADF) resistance-associated mt were found in genotypes B and C, respectively (ORLMV/ADF: 1495.000; 95% CI: 89.800–24,889.032; p < 0.001). Single-point mt associated to LMV and ADF resistance were detected in 59.9% of the tested children with rtV207M (38.0%) and rtN238T (9.9%) being the most frequent. Multiple-point mt were found only in 8 cases (5.6%): 6 children carried double mt (rtV207M + rtL229V; rtV207M + rtI233V; rtV207I + rtV207M × 2 cases; rtV207M + rtS213T; rtN238A + rtS256G) relating to LMV or/and ADF resistance and 3 children carried triple mt (rtL180M + rtM204I + rtN238T; rtV207M + rtS213T + rtS256G) or quadruple mt (rtL180M + rtM204V + rtV207I/M) for LMV-ADF resistance and Entecavir-reduced susceptibility. Our data indicate that significantly higher frequencies of LMV and ADF-associated mutations were found in treatment-naïve children infected with HBV genotypes B and C, respectively. The developed COLD-PCR method and obtained data may contribute to the development of suitable treatments for children with CHB.

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Profile of hepatitis B virus resistance mutations against nucleoside/nucleotide analogue treatment in Chinese patients with chronic hepatitis B

Virology Journal ◽

10.1186/1743-422x-10-313 ◽

2013 ◽

Vol 10 (1) ◽

pp. 313 ◽

Cited By ~ 6

Author(s):

Jun Lei ◽

Ying Wang ◽

Li-Li Wang ◽

Shao-Jun Zhang ◽

Wei Chen ◽

...

Keyword(s):

Chronic Hepatitis ◽

Hepatitis B Virus ◽

Hepatitis B ◽

Chronic Hepatitis B ◽

Virus Resistance ◽

Chinese Patients ◽

Resistance Mutations ◽

Nucleotide Analogue ◽

B Virus

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Genotyping of HBV and tracking of resistance mutations in treatment-naïve patients with chronic hepatitis B

Infection and Drug Resistance ◽

10.2147/idr.s135420 ◽

2017 ◽

Vol Volume 10 ◽

pp. 201-207 ◽

Cited By ~ 7

Author(s):

Sidelcina Rugieri Pacheco ◽

Maria Isabel Magalhães Andrade dos Santos ◽

Andreas Stocker ◽

Maria Alice Sant'Anna Zarife ◽

Maria Isabel Schinoni ◽

...

Keyword(s):

Chronic Hepatitis ◽

Hepatitis B ◽

Chronic Hepatitis B ◽

Resistance Mutations

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Fast and sensitive real-time PCR detection of major antiviral drug-resistance mutations in chronic hepatitis B patients using a predesigned panel of Locked-Nucleic-Acid TaqMan probes

Journal of Clinical Microbiology ◽

10.1128/jcm.00936-21 ◽

2021 ◽

Author(s):

Son V. Chu ◽

Son T. Vu ◽

Hang M. Nguyen ◽

Ngan T. Le ◽

Phuong T. Truong ◽

...

Keyword(s):

Drug Resistance ◽

Chronic Hepatitis ◽

Nucleic Acid ◽

Hepatitis B ◽

Real Time ◽

Chronic Hepatitis B ◽

Real Time Pcr ◽

Locked Nucleic Acid ◽

Resistance Mutations ◽

Drug Resistance Mutations

We developed a novel real-time PCR assay that simultaneously evaluates eleven major nucleos(t)ide antiviral (NA) drug-resistance mutations (mt) in chronic hepatitis B patients (CHB), including L180M, M204I/V, and V207M (lamivudine (LMV) resistance), N/H238A/T (Adefovir (ADF) resistance) which are circulating in Vietnam; T184G/L, S202I, M250V (entecavir (ETV) resistance), and A194T (tenofovir resistance) which have been recently reported in several studies across the globe. We detected drug-resistant mt in HBV samples using our predesigned panel of allele-specific locked-nucleic acid (LNA) probes. Our assay had a high sensitivity of 5% in a low HBV DNA population of ≥5 ×10 3 IU/mL and was validated in a cohort of 130 treatment-naive children and 98 NA-experienced adults with CHB. Single-point mt for LMV- and ADF-resistance were detected in 57.7% and 54.1% of the child and adult samples, respectively, with rtV207M (42.3%, children; 36.7%, adults) and rtN238T/A (15.4%, children; 16.3, adults) being the most frequent mt in these populations. Multiple-point mt including rtL180M-rtM204V- rtN238A and rtL180M-rtM204I were only identified in two children, resulting in LMV-ADF-resistance and reduced ETV susceptibility. In conclusion, this assay accurately identified the mt profile of children (98.4%) and adults (91.2%) with CHB, which is comparable to established methods. This fast and sensitive screening method can be used for the detection of major NA-resistant mt circulating in developing countries, as well as providing a model for the development of similar mt-detection assays, especially for use in non-hospitalised patients who need their results within half a day before starting treatment.

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