genotypic resistance profile of hepatitis b virus hbv in a large cohort of nucleos t ide CORD-Papers (Version 1)

Title: Genotypic resistance profile of hepatitis B virus (HBV) in a large cohort of nucleos(t)ide analogueexperienced Chinese patients with chronic HBV infection
Abstract: Summary. The study investigated the hepatitis B virus (HBV) genotypic resistance profile in 1803 nucleos(t)ide analogue (NA)experienced Chinese patients with chronic HBV infection. Serum HBV DNA was extracted and the reverse transcriptase region was analysed by a highsensitive direct PCR sequencing and verified by clonal sequencing if necessary. Drugresistant mutations were detected in 560 of the 1803 patients including 214 of 490 patients who received lamivudine (LAM) 35 of 428 patients who received adefovir (ADV) five of 18 patients who received telbivudine and 306 of 794 patients who received various sequential/combined NA therapies. ADVresistant mutations were detected in 36 of 381 patients who received LAM and then switchedto ADV in contrast to one of 82 patients who received ADV addon LAM. Entecavir (ETV)resistant mutations were detected not only in LAM and ETVtreated patients but also in LAMtreated ETVnave patients. Double mutations rtM204I and rtL180M were detected more frequently in genotype C than in genotype B virus and patients infected with this mutant had higher alanine transaminase levels than those infected with mutant containing the rtM204I substitution alone. Multidrugresistant HBV strains were identified in eight patients including two novel strains with mutational patterns rtL180M + A181V + S202G + M204V + N236T and rtL180M + S202G + M204V + N236T. The results provide new information on HBV genotypic resistance profiles in a large cohort of Chinese patients with chronic HBV infection and may have important clinical implication for HBV drug resistance management in China.
Published: 2010-09-06
Journal: J Viral Hepat
DOI: 10.1111/j.1365-2893.2010.01360.x
DOI_URL: http://doi.org/10.1111/j.1365-2893.2010.01360.x
Author Name: Liu Y
Author link: https://covid19-data.nist.gov/pid/rest/local/author/liu_y
Author Name: Wang C
Author link: https://covid19-data.nist.gov/pid/rest/local/author/wang_c
Author Name: Zhong Y
Author link: https://covid19-data.nist.gov/pid/rest/local/author/zhong_y
Author Name: Li X
Author link: https://covid19-data.nist.gov/pid/rest/local/author/li_x
Author Name: Dai J
Author link: https://covid19-data.nist.gov/pid/rest/local/author/dai_j
Author Name: Ren X
Author link: https://covid19-data.nist.gov/pid/rest/local/author/ren_x
Author Name: Xu Z
Author link: https://covid19-data.nist.gov/pid/rest/local/author/xu_z
Author Name: Li L
Author link: https://covid19-data.nist.gov/pid/rest/local/author/li_l
Author Name: Yao Z
Author link: https://covid19-data.nist.gov/pid/rest/local/author/yao_z
Author Name: Ji D
Author link: https://covid19-data.nist.gov/pid/rest/local/author/ji_d
Author Name: Wang L
Author link: https://covid19-data.nist.gov/pid/rest/local/author/wang_l
Author Name: Zhang L
Author link: https://covid19-data.nist.gov/pid/rest/local/author/zhang_l
Author Name: Wong V W S
Author link: https://covid19-data.nist.gov/pid/rest/local/author/wong_v_w_s
Author Name: Zoulim F
Author link: https://covid19-data.nist.gov/pid/rest/local/author/zoulim_f
Author Name: Xu D
Author link: https://covid19-data.nist.gov/pid/rest/local/author/xu_d
sha: 0f9c43aca920761d4e91a8e9a859a50a9408534c
license: no-cc
license_url: [no creative commons license associated]
source_x: Medline; PMC
source_x_url: https://www.medline.com/https://www.ncbi.nlm.nih.gov/pubmed/
pubmed_id: 21392168
pubmed_id_url: https://www.ncbi.nlm.nih.gov/pubmed/21392168
pmcid: PMC7167191
pmcid_url: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7167191
url: https://www.ncbi.nlm.nih.gov/pubmed/21392168/ https://doi.org/10.1111/j.1365-2893.2010.01360.x
has_full_text: TRUE
Keywords Extracted from Text Content: adefovir hepatitis B virus patients lamivudine Entecavir HBV Serum HBV DNA LAM ADVresistant rtL180M alanine transaminase LAM-treated ETV-naïve patients rtL180M + A181V rtM204I LAM-and nucleos(t)ide telbivudine ADV rtQ215S GQ402161 Red tenofovir · 10 4 HBeAg/anti-HBe telbivudine HBV DNA rtV214A nucleoside rtM204I/V. HBV DNA Serum ALT HCC entecavir rtI233V rtM204I/V rtL180M + M204V · 10 5 LAM-resistant rt250 rtM204I LAM [20] rt173 Ôgold liver patients rtM250I/L/ V · 10 6 manufacturerÕs [9] [10] [11] rtL217R MDR B rtA194T LdT rtM204V 1225-bp-long dATP rtN236T + A181T Serum samples serum ALT rtM204V + L180M ± V173L LAM-R/ETV-R DOWN1 UP4 DNA patient rtL80I Fosun Pharmaceutical Co. Hepatitis B virus Nonoptimal rt204 rt80 YMDD CHB-LC patients SARS coronavirus rtA181V LAM-and/or LAM people alanine transaminase Taq DNA polymerase · 10 3 IFN rtA181T rt84 [7] lamivudine GQ402162 serum HBV DNA ‡2000 hepatitis ADV adefovir hepatocellular carcinoma rtM250 amino acid rtL180M tube rtM204I + L180M ÔnewÕ ETV-naïve patients rtN236T anti-HBc GAA GAC AGC CHB-LC [13] [14] [15] [16] LAM switching-to ADV LAM-R anti-HBV NA MDR strains CHB LAM switching-to LdT SP6 ADV-R TDF ‡100 HBsAg rtV173L UP3 hepatitis B virus rtV84M rtA181T/S rtS202C/G/I 5¢-AGG LAM-experienced LdT. line hepatic inflammation Blue HBV HBsAg/anti-HBs NA rt180 Madison, WI ALT LAMresistant nucleotide lamivudine- HBeAg rtM204I + L180M ± V173L nucleos(t)ide ± rtL180M ± V173L rtL180M + A181V ETV
Extracted Text Content in Record: First 5000 Characters:The study investigated the hepatitis B virus (HBV) genotypic resistance profile in 1803 nucleos(t)ide analogue (NA)-experienced Chinese patients with chronic HBV infection. Serum HBV DNA was extracted, and the reverse transcriptase region was analysed by a high-sensitive direct PCR sequencing and verified by clonal sequencing if necessary. Drug-resistant mutations were detected in 560 of the 1803 patients, including 214 of 490 patients who received lamivudine (LAM), 35 of 428 patients who received adefovir (ADV), five of 18 patients who received telbivudine and 306 of 794 patients who received various sequential/combined NA therapies. ADVresistant mutations were detected in 36 of 381 patients who received LAM and then switched-to ADV in contrast to one of 82 patients who received ADV add-on LAM. Entecavir (ETV)-resistant mutations were detected not only in LAM-and ETV-treated patients but also in LAM-treated ETV-naïve patients. Double mutations rtM204I and rtL180M were detected more frequently in genotype C than in genotype B virus, and patients infected with this mutant had higher alanine transaminase levels than those infected with mutant containing the rtM204I substitution alone. Multidrug-resistant HBV strains were identified in eight patients, including two novel strains with mutational patterns rtL180M + A181V + S202G + M204V + N236T and rtL180M + S202G + M204V + N236T. The results provide new information on HBV genotypic resistance profiles in a large cohort of Chinese patients with chronic HBV infection and may have important clinical implication for HBV drug resistance management in China. Hepatitis B virus (HBV) chronic infection afflicts about 350 million people worldwide, of whom 93 million live in China [1, 2] . Morbidity and mortality in chronic hepatitis B (CHB) are linked to persistent viral replication and evolution to CHB-related liver cirrhosis (CHB-LC) or hepatocellular carcinoma (HCC) [3] . In China, four nucleos(t)ide analogues (NA), i.e. lamivudine (LAM), adefovir (ADV), entecavir (ETV) and telbivudine (LdT) are currently approved for the treatment of HBV infection, whilst recently developed tenofovir (TDF) is still unavailable. Treatment of CHB and CHB-LC is aimed at suppressing viral replication to the lowest possible level, and thereby halting the progression of liver disease. However, viral resistance is the main drawback of long-term antiviral therapy [4] [5] [6] . Suboptimal treatment regimens and drug-resistant viral infection can increase the incidence of drug resistance and may favour the selection of multidrugresistant (MDR) HBV [7, 8] . The resistance mutations are located in the reverse transcriptase (RT) region of the HBV polymerase gene. The rtM204I and rtM204V are classic LAM-resistant mutations and often coexist with compensatory mutations (rtV173L and rtL180M) [9] [10] [11] . The rtN236T and rtA181V are two well-recognized ADV-resistant mutations [12, 13] , and some purported mutations such as rtV84M, rtV214A, rtQ215S, rtL217R and rtI233V may reduce susceptibility to ADV, although these are still controversial [13] [14] [15] [16] . Substitutions in rtT184, rtS202 or rtM250 in conjunction with LAMresistant mutations result in ETV resistance [17] [18] [19] . Cross-resistance (usually rtM204I) also exists between LdT and LAM [20] . In addition, rtA181T seems to be an atypical substitution associated with LAM and ADV selection and may reduce the typical extent of virologic breakthrough [21] . Genotypic antiviral resistance is designated by the presence of unique nucleotide and corresponding deduced amino acid mutations in the drug target gene that have been previously demonstrated to be associated with antiviral resistance. The incidence of genotypic resistance is related to viral factors, host factors and treatment characteristics and is also affected by the methods used for detection of resistance mutations and the patient population being studied [22] . Several methods have been used for typing HBV genetic drug-resistant mutations, each with individual advantages and disadvantages [1, 22] . Direct polymerase chain reaction (PCR) sequencing is the most popular method owing to the abundant information it provides. However, it is also considered less sensitive for typing samples with low viral load (<2000 IU/mL) and minor mutant subpopulations (<20%) [22] . As more antiviral strategies become available for the treatment of CHB and CHB-LC, the risk and patterns of resistant and cross-resistant emergence are diverse. Nonoptimal strategies based on the sequential use of NA increase the development of MDR strains [23] . Knowledge on the incidence and patterns of drug-resistant mutants is valuable for clinicians and would assist clinical monitoring and management of the resistance. To date, data are largely derived from a few clinical trials and cohorts with limited drug resistance profiling. This study is intended to investigate population-based profiles of HBV genotypic resi
Keywords Extracted from PMC Text: rtQ215S rtM204I/V rtV214A CHB‐LC patients one‐tube add‐on 5′‐AGT CAG GAA GAC AGC rtI233V rtL217R HBV DNA ETV rtN236T YMDD chi‐square tests INNO‐LiPA 54–75 B ADV‐R rtL180M + A181V LdT‐resistant Nonoptimal Taq DNA polymerase alanine transaminase 5′‐AGG TGA AGC GAA GTG CAC AC‐3′ LdT. LAM switching‐to ADV MDR strains patient LAM switching‐to LdT DNA adefovir line rtA181V rtM204I serum HBV DNA long‐term rtM204V LAM‐R B, C ALT tube ETV‐naïve patients wild‐type Serum rtA194T rt84 hepatic inflammation rtA181T/V CGC AGT ATG rtN236T + A181T entecavir UP3 rtM250I/L/V dATP LAM rtM204I + L180M patients nucleotide Serum samples rtM204I/V. HBsAg/anti‐HBs B and D ≥2000 MDR HBV strains CHB Hepatitis B virus hepatocellular carcinoma B (44.6%/55.4%vs 19.4%/80.6 anti‐HBV NA tenofovir rt204 TDF telbivudine Fosun Pharmaceutical Co. HBV rtL180M serum ALT rtL180M + M204V ADV rt80 × GQ402162 NA wild‐type HBV ≥100 rtV173L people rtV84M rt180 LdT lamivudine rtA181T/S Madison, WI GQ402161 Coexist‐R HCC nucleoside less‐sensitive 's rt173 MDR ≥20 nucleos(t)ide HBeAg [12, 13] DOWN1 LAM [20] [7] liver × 106 rtA181T hepatitis HBsAg rtS202C/G/I rtL180M ± V173L ADV add‐on LAM‐experienced rtM250 rtM204I + L180M ± V173L coronavirus Patients immunoglobulin LAM‐ Coexist‐R samples amino acid CHB‐LC rtL80I rtM204V + L180M ± V173L SP6 Hepatitis B virus genotypes C rt250
Extracted PMC Text Content in Record: First 5000 Characters:Hepatitis B virus (HBV) chronic infection afflicts about 350 million people worldwide, of whom 93 million live in China [1, 2]. Morbidity and mortality in chronic hepatitis B (CHB) are linked to persistent viral replication and evolution to CHB‐related liver cirrhosis (CHB‐LC) or hepatocellular carcinoma (HCC) [3]. In China, four nucleos(t)ide analogues (NA), i.e. lamivudine (LAM), adefovir (ADV), entecavir (ETV) and telbivudine (LdT) are currently approved for the treatment of HBV infection, whilst recently developed tenofovir (TDF) is still unavailable. Treatment of CHB and CHB‐LC is aimed at suppressing viral replication to the lowest possible level, and thereby halting the progression of liver disease. However, viral resistance is the main drawback of long‐term antiviral therapy [4, 5, 6]. Suboptimal treatment regimens and drug‐resistant viral infection can increase the incidence of drug resistance and may favour the selection of multidrug‐resistant (MDR) HBV [7, 8]. The resistance mutations are located in the reverse transcriptase (RT) region of the HBV polymerase gene. The rtM204I and rtM204V are classic LAM‐resistant mutations and often coexist with compensatory mutations (rtV173L and rtL180M) [9, 10, 11]. The rtN236T and rtA181V are two well‐recognized ADV‐resistant mutations [12, 13], and some purported mutations such as rtV84M, rtV214A, rtQ215S, rtL217R and rtI233V may reduce susceptibility to ADV, although these are still controversial [13, 14, 15, 16]. Substitutions in rtT184, rtS202 or rtM250 in conjunction with LAM‐resistant mutations result in ETV resistance [17, 18, 19]. Cross‐resistance (usually rtM204I) also exists between LdT and LAM [20]. In addition, rtA181T seems to be an atypical substitution associated with LAM and ADV selection and may reduce the typical extent of virologic breakthrough [21]. Genotypic antiviral resistance is designated by the presence of unique nucleotide and corresponding deduced amino acid mutations in the drug target gene that have been previously demonstrated to be associated with antiviral resistance. The incidence of genotypic resistance is related to viral factors, host factors and treatment characteristics and is also affected by the methods used for detection of resistance mutations and the patient population being studied [22]. Several methods have been used for typing HBV genetic drug‐resistant mutations, each with individual advantages and disadvantages [1, 22]. Direct polymerase chain reaction (PCR) sequencing is the most popular method owing to the abundant information it provides. However, it is also considered less sensitive for typing samples with low viral load (<2000 IU/mL) and minor mutant subpopulations (<20%) [22]. As more antiviral strategies become available for the treatment of CHB and CHB‐LC, the risk and patterns of resistant and cross‐resistant emergence are diverse. Nonoptimal strategies based on the sequential use of NA increase the development of MDR strains [23]. Knowledge on the incidence and patterns of drug‐resistant mutants is valuable for clinicians and would assist clinical monitoring and management of the resistance. To date, data are largely derived from a few clinical trials and cohorts with limited drug resistance profiling. This study is intended to investigate population‐based profiles of HBV genotypic resistance in Chinese patients, with an improved direct PCR sequencing assay. Serum samples were collected from 1803 CHB and CHB‐LC patients who visited Beijing 302 Hospital during July 2007–March 2009. The standard for diagnoses of CHB and CHB‐LC was based on the Chinese Management Scheme of Diagnostic and Therapy Criteria of Viral Hepatitis [24] and have been described elsewhere [25, 26]. The male/female ratio was 1524/279. Average age was 37.2 ± 13.6 years. At the time of sampling for HBV genotyping, all patients were HBsAg positive, and 1203 (66.7%) patients were HBeAg positive. The median (Q1, Q3) of the alanine transaminase (ALT) level was 38 (24, 70) U/L; and the median (Q1, Q3) of the HBV DNA level was 2.8 × 104 (1.7 × 103, 7.5 × 105) IU/mL. All patients had received anti‐HBV NA (LAM, ADV, ETV and LdT) monotherapy, combination or sequential therapy for a minimum of 3 months. Written informed consents for the analysis were obtained from every patient. The study was approved by the ethics committee of Beijing 302 Hospital. Serum ALT, HBsAg/anti‐HBs, HBeAg/anti‐HBe, anti‐HBc and other biochemical and serological markers, as well as HBV DNA level were routinely measured or detected in the Central Clinical Laboratory of the Beijing 302 Hospital. HBV DNA level was determined by real‐time quantitative PCR (qPCR) (Fosun Pharmaceutical Co., Ltd., Shanghai, China) with a lower detection limit of 100 IU/mL (≈500 copies/mL). Hepatitis B virus gene fragment (nt 54–1278) encompassing the complete RT gene was amplified by nested PCR. The sense and antisense primers for the first‐round PCR were 5′‐AGT CAG GAA GAC AGC CTA CTC C‐3′ (
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