DNA, Viral ; analysis ; Drug Resistance, Viral ; Hepatitis B ; drug therapy ; immunology ; virology ; Humans ; Viral Load

Drug Resistance, Viral ; Hepatitis B virus ; drug effects

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

Antiretroviral Therapy, Highly Active ; China ; Drug Resistance, Viral ; HIV ; drug effects ; HIV Infections ; drug therapy ; Humans

Highly active antiretroviral combination therapy significantly reduced the mortality, but in the high-speed copying, high genetic variation and drug selection pressure under the effect of the increasingly serious problem of drug resistance greatly weakened the role of HAART inhibit viral replication and reduce antiviral treatment. This paper reports the latest trends in HIV drug-resistance in order to develop anti-HIV drugs in clinical programs, research and development of new guidance anti-HIV-1 strategy to bring guidance.
Anti-HIV Agents ; pharmacology ; Drug Discovery ; Drug Resistance, Viral ; HIV ; drug effects ; enzymology ; Humans ; Internationality

OBJECTIVETo evaluate the amplification rate and the lowestlower detection limit of an in-house HIV-1 Drug resistant (HIVDR) genotyping test.

METHODSA total of 30 plasma samples were selected, which covered all major HIV-1 subtypes predominating prevailing in China (B', CRF07_BC, CRF01 _AE). The viral loads of the 30 selected samples were detected in triplicate by Easy Q method and the average values were taken as the viral loads of the samples. Each sample was diluted to the concentration of > 1000 copies/ml, 401-1000 copies/ml, 101-400 copies/ml, 50-100 copies/ml and < 50 copies/ml with HIV-negative plasma. After extraction of nucleic acids, RT-PCR and nested PCR amplification were performed, the efficiency of amplification of each subtype and the minimum detection limit were determined statistically based on the PCR results.

RESULTSThe viral loads of the selected samples ranged from 2.03 x 10(2)-5.92 x 10(4) copies/ml. The sample of 50-1000 copies/ml have a high amplification rate (86%).

CONCLUSIONThe In-house method for HIV-1 drug resistance genotyping has a high sensitivity with a high successful amplification rate, especially in the samples with low viral load. This method can be used to the detection of drug-resistant virus and to provide scientific data to treatment options for patients.

China ; Drug Resistance, Viral ; Genotype ; HIV-1 ; classification ; drug effects ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; Viral Load

OBJECTIVETo find out the mechanism of drug resistance by detecting the mutations of HIV RNA in patients who failed in the anti-HIV therapy, to direct the clinical use of anti-HIV drugs and to complement the existing drug resistant database.

METHODSHIV RNA and DNA were extracted from the plasma of 10 HIV-infected patients who developed drug resistance in the Clinic of AIDS, Ruhr University, Bochum, Germany. Then HIV-RNA was amplified in the reverse transcriptase (RT) and protease regions by polymerase chain reaction (PCR). After purified, the PCR products was sequenced. The acquired sequences were compared with the international standard strain HXB2CG and the resistant database of Stanford University.

RESULTSSome mutations were found to cause the corresponding resistance to certain drugs and were consistent with the clinical results. Some mutations existed in some patients, such as V179I in RT and K20T, K20I in protease, which hadn't been reported in the resistant database of Stanford University yet.

CONCLUSIONPatients who fail in HAART have different mutations in RT and protease regions. Mutations such as V179I in RT and K20T, K20I etc in protease may be related to drug resistance.

Adult ; Antiretroviral Therapy, Highly Active ; Drug Resistance, Viral ; HIV Infections ; drug therapy ; virology ; Humans ; RNA, Viral ; blood ; Treatment Failure

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