Humans ; Mutation/genetics* ; HIV Infections/drug therapy* ; Drug Resistance ; Treatment Failure ; China ; Drug Resistance, Viral/genetics*

Objective: To understand immune escape mutation, drug resistance mutation, and genome evolution information of HBV genome sequence in China. Methods: The whole genome sequence information of HBV in China submitted in GenBank from 1998 to 2021 was selected as the object for analysis. MAFFT method was used for cluster analysis. Analysis of immune escape and drug-resistant mutations was performed using the online tool Gen2pheno. The BEAST 1.10.4 was used for analysis the time evolution of HBV sequences. Results: A total of 5 426 sequences were included in the dataset and distributed in 19 provinces of China. Type C accounted for the highest proportion (59.1%, 3 211/5 426), followed by type B (33.7%, 1 833/5 426). Immune escape mutations were found in 764 sequences (14.1%, 764/5 426). At least one reverse transcriptase region mutation occurred in 98.1% of the sequences. The evolutionary roots of most HBV sequences in China date from around 1801 AD. Conclusion: HBV-resistant mutation rate is high in China. HBV genomes evolve slowly.
China/epidemiology* ; DNA, Viral/genetics* ; Drug Resistance, Viral/genetics* ; Genome, Viral ; Genotype ; Hepatitis B virus/genetics* ; Humans ; Mutation

OBJECTIVETo develop a high-throughput clinical method on drug-resistance gene mutations of HBV using MALDI-TOF-MS.

METHODUsing MassArray Assay Design software designed the iPLEX primers and followed the iPLEX instruction for amplification, SAP reaction, primer extenction, desalination, dispensing, MALDI-TOF-MS screening and data analysis of the gene mutation locus. 138 serum samples of chronic HBV patients with single drug-resistance or multiple drug-resistance on Lamivudin, adefovi, Entecavir were detected.

RESULTThe HBV gene mutation platform was successfully developed and applied on the high-throughput dectection of clinical serum samples. It was also a high throughput assay which could be used to detect for more than 138 samples once. The MALDI-TOF-MS technology and the DNA sequencing simultaneously examine 33 samples, in which result of 10 sample is inconsistent, the including 2 samples by MALDI-TOF-MS technology has not tested, 1 sample has 2 inconsistent mutations.

CONCLUSIONDetection of HBV gene mutations using MALDI-TOF-MS is highly-sensitive, highly-accurate, high-throughput, fast achieved and suitable to use in the diagnosis and monitoring of HBV.

DNA, Viral ; genetics ; Drug Resistance ; genetics ; Drug Resistance, Viral ; drug effects ; genetics ; Hepatitis B virus ; drug effects ; Mass Spectrometry ; Polymorphism, Single Nucleotide

Drug Resistance, Viral ; genetics ; Hepatitis B virus ; drug effects ; genetics ; Mutation

Drug Resistance, Viral ; genetics ; Hepatitis B virus ; drug effects ; genetics ; Mutation ; Nucleosides ; pharmacology

OBJECTIVESince the advent in 2004 of highly active antiretroviral therapy (HAART) in Liaoning, a dramatic improvement had been seen in the number of patients attaining undetectable viral loads (92/104), but the extent of mutation diversity on human immunodeficiency virus 1 (HIV-1) and the prevalence of drug resistance had remained elusive. This study aimed to analyze both HIV-1 mutation profiles and prevalence related to antiretroviral resistance following therapeutic failure.

METHODSA total of 104 blood samples circling Liaoning from HAART-treated between 2004 and 2008 were studied. Patients' CD(4)(+) T-cell count and viral load were determined. HIV-1 pol (PR and part of RT) gene fragments were amplified from patients' plasma by reverse transcriptase polymerase chain reaction (RT-PCR) and nest-PCR, subsequently sequenced and analyzed.

RESULTSCD(4)(+) T cell numbers and viral replication capacity were assessed. 88.4% (92/104) of the patients were successful after initial non-suppressive NRTI & NNRTI-based HAART regimens. Subjects on non-nucleoside reverse transcriptase inhibitor (NNRTI) regimens developed more (6/104) drug-resistance mutations than those on nucleoside reverse transcriptase inhibitor (NRTI) regimens did (5/104). No protease-inhibitor (PI) drug resistance mutations developed. The whole rate of drug resistance mutations was about 6.73%. Subjects developing NNRTI-resistance (NNRTI-R) seemed more likely to develop drug-resistant viremia than with NRTI-based HAART.

CONCLUSIONThis finding might have implications in which that the prevalence of drug-resistance mutations was low but remained risk of transmission in HIV-infected therapeutic failure. Meanwhile, data from the present study showed that there was a high frequency of primary mutations, which offered resistance to nrti and nnrti. Monitoring patients with treatment failure seems an important tool in helping the physicians to improve their treatment schedule and to carry out epidemiological surveillance programs.

Acquired Immunodeficiency Syndrome ; drug therapy ; epidemiology ; virology ; China ; epidemiology ; Drug Resistance, Viral ; genetics ; HIV-1 ; drug effects ; genetics ; Humans ; Molecular Epidemiology ; RNA, Viral ; genetics ; Viral Load

OBJECTIVETo compare the concordance in predicting genotype HIV-1 drug resistance between In-house method and TRUGENE(TM) or ViroSeq(TM) method.

METHODS25 international proficiency testing (PT) samples received from 2009 to 2013 were detected by three methods, then pairwise comparison results was analyzed to validate their concordance on drug resistance mutation and drug resistance report. To further confirm the results, another 15 serum specimens were detected by In-house and TRUGENE(TM) methods, then compared their results concordance.

RESULTSThe evaluation of the consistency of resistance-associated mutations showed that for 25 PT samples, the consistency was 99.42% (2933/2950) in testing drug resistance mutation sites among the three methods; all pairwise comparison Kappa values >0.81(P < 0.01). The evaluation of the consistency of drug resistance test showed that inconsistent comparison results mainly concentrated in the four nucleoside reverse transcriptase inhibitors zidovudine (AZT), didanosine (ddI), stavudine (d4T), abacavir (ABC), and the inconsistencies were mainly minor. Where the minor inconsistencies between In-house and ViroSeq(TM) methods were 28% (7/25), 28% (7/25), 16% (4/25) and 20% (5/25), respectively. And the inconsistencies between In-house and TRUGENE(TM) method were separately 44% (11/25), 28% (7/25), 36% (9/25) and 28% (7/25);while the inconsistencies between TRUGENE(TM) and ViroSeq(TM) method were separately 24% (6/25), 8% (2/25), 28% (7/25) and 8% (2/25). AZT in the comparison between In-house and ViroSeq(TM) methods, ddI, d4T, ABC and TDF in the comparison between In-house and TRUGENE(TM) methods were moderately consistent (weighted Kappa values were separately 0.54, 0.44, 0.52, 0.42, 0.59, all the P value <0.01). The other compared results were all highly-consistent (weighted Kappa values were 0.61-0.80) or extremely high-consistent (weighted Kappa values were >0.80). For 15 serum specimens, 99.55% (1762/1770) drug resistance mutation sites could be detected by the two methods, the difference about drug results concentrated in AZT, ddI, d4T and ABC.

CONCLUSIONThe In-house genotyping system had a high concordance with commercial TRUGENE(TM) or ViroSeq(TM) genotyping system.

Drug Resistance, Viral ; genetics ; Genes, Viral ; Genotype ; HIV-1 ; drug effects ; genetics ; isolation & purification ; Humans ; Reagent Kits, Diagnostic

Adenine ; analogs & derivatives ; pharmacology ; Drug Resistance, Viral ; genetics ; Genome, Viral ; Hepatitis B virus ; drug effects ; genetics ; Organophosphonates ; pharmacology ; Plasmids ; Viral Proteins ; genetics

OBJECTIVETo analyze the genome mutations of HIV-1 gag, pol and env genes from HIV-infected paid blood donors in rural central China.

METHODSDNA was extracted from peripheral blood mononuclear cells, gag (p17-p24), pol (PR-RT), env (C2-V5) genes were amplified by nested polymerase chain reaction (PCR), purified products were sequenced, and sequence data was analyzed by MEGA5.0 soft wares.

RESULTSTwenty-three samples were subtype B, two samples were recombinant of subtype B and subtype C, one sample was recombinant of subtype CRF01_AE and subtype B. PI major resistance mutations were not found in the PR region. M184V, K101E and G190A were detected in the RT region, respectively.

CONCLUSIONSubtype B was the major HIV circulating genetic forms in this area. Most strains were sensitive to high active anti-retroviral therapy (HARRT). 91.7% V3 loop tip motifs of X4-tropic strains was GPGR. It showed that GPGR might be associated with accelerate disease progression to AIDS.

Blood Donors ; Drug Resistance, Viral ; genetics ; Genes, pol ; Genotype ; HIV-1 ; classification ; genetics ; Humans ; Phylogeny

OBJECTIVETo observe the distribution of HBV variants resistant to lamivudine and their relation to clinical manifestations of chronic hepatitis.

METHODSUsing direct sequencing, YMDD (tyrosine-methionine-aspartate-aspartate) variants in patients with chronic HBV were detected before and during treatment with lamivudine. A statistical analysis of the distribution of HBV strains resistant to lamivudine was performed.

RESULTFour variant strains existed in patients before lamivudine treatment, 128 variant resistant strains were noted after 6 mouths of lamivudine treatment including 42 YVDD (valine) variants, 20 YIDD (isoleusine) variants and 66 non-YMDD variants. According to the hepatitis severity, 8 patients were mild, 108 moderate and 12 severe. Viral loading was higher and clinical types were more severe in no-YMDD variants.

CONCLUSIONVariant strains including strains resistant to lamivudine exist naturally before lamivudine treatment, but lamivudine-resistant ones become more dominant after treatment. Liver inflammation is more severe in non-YMDD group.

Antiviral Agents ; pharmacology ; Drug Resistance, Viral ; Genetic Variation ; Hepatitis B virus ; drug effects ; genetics ; Lamivudine ; pharmacology