No Abstract Available.
Genes, pol* ; Korea* ; Leukemia Virus, Bovine* ; Proviruses*

No abstract available.
Genes, pol* ; HIV-1 ; Humans* ; RNA-Directed DNA Polymerase ; Zidovudine*

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 investigate the drug resistance of HIV patients to the HIV-1 CRF01_AE and CRF07_BC strains in Sichuan province during 2010 to 2013.

METHODS1.5 ml of plasma were collected from AIDS patients who had been receiving anti-retroviral treatment for over 6 months but still had a HIV-1 virus load of over 1 000 copies/ml from January 1, 2010 to December 31, 2013 in Sichuan province. Genetic analysis of the HIV-1 pol gene was performed using self-established method, and patients with a positive drug-resistant HIV-1 pol gene mutation were included. HIV-1 poly gene was successfully sequenced for a total of 1 213 patients. Drug resistance of different HIV-1 strains was compared with χ2 test or Fisher exact test.

RESULTS558 cases (46.0%) of the 1 213 successfully sequenced patients were infected by HIV-1-strains with drug-resistant mutations, including 327 cases (58.6%) infected by CRF01_AE strain, 126 (22.6%) by CRF07_BC strain, 46 (8.2%) by CRF08_BC strain, 33 (5.9%) by B strain, 4 (0.7%) by C strain, 1 (0.2%) by CRF02_AG strain, and 21 (3.8%) by unidentified strains. Drug-resistant mutation analysis revealed that L33, F116, L74, Q151, and T69 resistance mutations occurred only in the CRF01_AE strain, while A71, K43, and Q58 resistance mutations occurred only in the CRF07_BC strain; in nuclear nucleoside reverse transcriptase inhibitors (NRTIs) and non nucleoside reverse transcriptase inhibitors (NNRTIs), CRF01_AE subtype strains showed highly resistant rate were higher than CRF07_BC, CRF08_BC and B subtype strains, with the differences were statistically significant (P<0.05).

CONCLUSIONThe drug-resistant HIV-1 strains in Sichuan mainly included the CRF01_AE and CRF07_BC strains, which had different resistance mutations.

Base Sequence ; Drug Resistance, Viral ; Genes, pol ; HIV Infections ; HIV-1 ; Humans ; Mutation ; Reverse Transcriptase Inhibitors ; Viral Load

OBJECTIVETo investigate the distribution and proportion of subtypes of pol gene in HIV-1 epidemic strains in Guangxi Autonomous Region.

METHODS152 HIV-1 patients were enrolled from 11 cities in Guangxi Autonomous Region from 2010 to 2012 by convenient sampling. Inclusion criterias were listed as the fdlowing: HIV-1 infection was confirmed by Western blot, HIV-1 viral load >1 000 copies/ml, > 18 year-old, and without any serious illnesses. 5 ml of peripheral blood samples were obtained from each patient. The viral RNA was isolated from plasma and used for amplification of full-length pol gene by nested RT-PCR. The amplified products were sequenced. After editing and modification, all sequences were characterized for preliminary subtyping by genotyping and confirmed with phylogenetic tree constructed by MEGA 5.03 software. The recombinant identification of 2 unknown recombinant strains was determined by RIP and jpHMM at GOBICS.

RESULTSAmong 152 patients, 137 full-length pol genes were successfully amplified and 127 HIV-1 subtypes were identified. The distribution and proportion of subtypes was summarized as the following 71 cases of CRF01_AE, accounting for 55.9% (71/127), 38 CRF08_BC, 29.9% (38/127), 13 CRF07_BC, 10.2% (13/127), and 3 B (B'), 2.4% (3/127), 2 unknown recombinant strains, 1.6% (2/127). In 11 cites of Guangxi Autonomous Region, subtype CRF01_AE was the dominant strain. Among heterosexual transmitted patients and drug abusers, the proportions of subtype CRF01_AE were 67.4% (58/86) and 34.1% (14/41), respectively. There was a significance different in the distribution of CRF01_AE in different routes of transmission (χ(2)=15.07, P<0.001). In age 21- 35, age 36- 60 and age>60 groups, the proportions of CRF01_AE was 43.6% (17/39), 57.6% (38/66), 77.3% (17/22), and CRF08_BC was 43.6% (17/39), 28.8% (19/66), 9.1% (2/22), respectively, the difference in proportions was significant(χ(2)=8.48, P= 0.014). The patterns of two unknown recombinant strains were found to be CRF01_AE/B (B') and CRF01_AE/C/B(B'), respectively.

CONCLUSIONCRF01_AE was the dominant HIV-1 subtype in Guangxi Autonomous Region from 2010 to 2012, with heterosexual transmission as its main spreading route. The two unknown recombinant strains in Guangxi Autonomous Region were reconstructed by subtype CRF01_AE and CRF_BC.

Blotting, Western ; China ; epidemiology ; Cities ; Drug Users ; Genes, pol ; Genotype ; HIV Infections ; epidemiology ; transmission ; virology ; HIV-1 ; genetics ; Humans ; Phylogeny ; Polymerase Chain Reaction ; RNA, Viral ; blood ; pol Gene Products, Human Immunodeficiency Virus ; genetics

Objective: To understand the epidemiological characteristics of one large HIV molecular transmission cluster in Jiaxing city, Zhejiang province, 2017 in order to select those people under high-risk and providing basis for programs on prevention. Methods: During 2017, newly diagnosed HIV/AIDS cases in this city were recruited. Plasma samples were collected from subjects, followed by RNA extraction, RT-PCR and nest-PCR for pol gene amplification, before being sequenced and aligned. Mega 6.0 software was used to construct phylogenetic tree, and Cytoscape 3.6.0 software was used to identify HIV molecular transmission clusters. Cases within the large transmission clusters were investigated, using a field-epidemiology-questionnaire. Data related to socio-demographics and previous sexual behaviors were collected and EpiData 3.0 and SPSS 20.0 software were used. Results: In the large transmission cluster with subtype identified as CRF07_BC, in Jiaxing, 2017, 26 cases of the total 30 cases were investigated. A total of 80.8% (21/26) could be identified as newly infected within the last two years and 30.8%(8/26) could be identified as newly infected within the last one year, including 22 cases infected locally. Among several infected cases who were at age 45 years or older, they admitted that they had experienced unprotected sexual contacts in local city for long time and having had more than 10 disclosed sexual contacts within the last two years at the local venues. Conclusions: This molecular cluster had been formed and scaled up quickly in recent two years, it has played an important role in promoting and scaling up the HIV transmission. Three cases identificed as high risk played an importantrde role in scaling up this cluster.
Amplified Fragment Length Polymorphism Analysis ; China/epidemiology* ; Genes, pol ; Genotype ; HIV Infections/transmission* ; HIV-1/isolation & purification* ; Humans ; Molecular Epidemiology ; Phylogeny ; Polymerase Chain Reaction ; RNA, Viral/blood* ; Sexual Behavior ; pol Gene Products, Human Immunodeficiency Virus

OBJECTIVETo study the epidemic status of human immunodeficiency virus type 1 (HIV-1) subtypes in Shenzhen and to study their transmission source and routes.

METHODSHIV-1 env and gag genes were amplified by nested PCR from uncultured peripheral blood mononuclear cells (PBMCs) obtained from 122 HIV-1 carriers confirmed in Shenzhen. The C2-V3 region (about 450 bp) of HIV-1 env and P17/ P24 region were sequenced.

RESULTSAmong 122 samples, 6 HIV-1 strains including 3 circulating recombinant forms (CRFs) of CRF01_AE, CRF08_BC, CRF07_BC and 3 subtypes of B', B, C were found in Shenzhen, and the percentages were 45.1% (55/122) for CRF01_AE, 31.1% (38/122) for CRF08_BC, 6.6% (8/122) for CRF07_BC, 14.8% (18/122) for B' subtype, 1.6% (2/122) for B subtype, and 0.8% (1/122) for C subtype. The intragroup genetic distances were (4.455 +/- 1.478)%, (2.997 +/- 1.345)%, (4.380 +/- 2.024)%, (5.186 +/- 2.487)%, and (4.869 +/- 2.638)%, respectively. In comparison with the sequence of respective international strains 01AE. TH. 90. CM240, 97CNGX-9F, CN. 97. C54A, B. US. 83. JRFL, and RLA2, the genetic distances were (5. 228 +/- 0.823)%, (3.634 +/- 1.073)%, (4.233 +/- 1.119)%, (4.950 +/- 2.564)%, and (5.795 +/- 2.198)%, respectively. The major subtypes found in injection drug users (IDUs) were CRF07_BC, CRF08_BC, and CRF01_AE strains. CRF01_AE and B' strains were epidemic mainly in sexual workers.

CONCLUSIONThere are 3 HIV-1 subtypes (B', B, C) and 3 CRFs (CRF01_AE, CRF08_BC, CRF07_BC) epidemics in Shenzhen. The predominant subtypes varies among different transmission routes. While CRF01_AE is predominant among sexual workers, CRF08_BC and CRF01_AE are major subtypes among IDU population.

Adolescent ; Adult ; China ; epidemiology ; Female ; Genes, env ; genetics ; Genes, gag ; genetics ; Genes, pol ; genetics ; HIV Infections ; epidemiology ; HIV-1 ; genetics ; Humans ; Male ; Middle Aged ; Molecular Epidemiology ; Polymerase Chain Reaction

This study aims to compare the influence of different genes to the results of HIV-1 subtype B' phylogenetic analyses. We first split 47 near full-length genome sequences of subtype B' into different regions (gag, pol, vif, vpr, vpu, env, nef), which derived from various risk populations and geographic regions from Thailand, Myanmar and China from published studies. Phylogenetic analyses were performed to each region obtained. The phylogenetic results of different regions were compared to that of the near full-length genome sequences. The pol gene was found to have the lowest diversity and evolutionary rate, and could repeat the phylogenetic results by using near full-length genome sequences. Although the env gene has the highest diversity and evolutionary rate, it could not achieve the similar results. This study compared the influence to the results of HIV-1 subtype B' phylogenetic analyses by using different genes and laid foundation for further molecular survey and analyses of the transmission of subtype B' in China.
Amino Acid Sequence ; China ; epidemiology ; Cluster Analysis ; Genes, Viral ; genetics ; HIV-1 ; classification ; genetics ; Molecular Epidemiology ; Molecular Sequence Data ; Phylogeny ; pol Gene Products, Human Immunodeficiency Virus ; chemistry ; genetics

OBJECTIVETo understand the HIV-1 subtype diversity and transmission characteristics in men having sex with men (MSM) in Zhejiang province.

METHODSA total of 233 newly diagnosed as HIV-1 positive patients in 2011 were screened out by BED capture enzyme immunoassay (BED-CEIA). Among them, 107 eligible subjects were enrolled for further molecular epidemiological study. Viral RNA was extracted from plasma samples and followed by reverse transcription PCR and nested PCR for amplification of pol gene fragments, sequencing, and bioinformatics analysis.

RESULTSThere were no statistically significant differences regarding the social demographic distribution between the subjects under study and those recently infected MSM population. The rate of success for sequence acquisition was 94.4% (101/107). The highest proportion of subtype was CRF01_AE (62.4%), followed by CRF07_BC (31.7%) and with three cases of subtype B, one case of CRF55_01B and two cases of unique recombinant form (CRF01_AE/B and CRF01_AE/CRF07_BC). The phylogenetic trees were mainly divided into CRF01_AE cluster 1, cluster 2 and CRF07_BC cluster 3. The strains located in Hangzhou were diffused in the branches of phylogenetic tree. 10 transmission clusters were found, in which 80% involved two or more regions and 90% was associated with patients residing in Hangzhou. Three surveillance drug resistance mutations (M46I, T215S and G190A) were found in three samples (each sample harbored only one resistance mutation). The overall rate of transmitted drug resistance (TDR) was 2.97%.

CONCLUSIONThe increasing complexity of HIV was noticed in MSM in Zhejiang province. However, the prevalence of TDR was low. Cross-regional HIV transmission in MSM was common, which inferred from the study. Hangzhou might play a central regional role in the intra-provincial spread of HIV, to form an interwoven complex network in the MSM population.

China ; epidemiology ; Demography ; Drug Resistance, Viral ; Genes, pol ; HIV Infections ; genetics ; transmission ; HIV-1 ; classification ; Homosexuality ; Humans ; Immunoenzyme Techniques ; Male ; Phylogeny ; Prevalence ; RNA, Viral

OBJECTIVETo construct and compare the immunogenicities of DNA vaccines expressing pol genes derived from B`/C and A/E recombinant subtypes of HIV-1 in China.

METHODSTwo DNA vaccines were constructed by inserting the codon optimized pol genes derived from B'/C and A/E subtypes of HIV-1 into mammalian expression vector pSV1.0. In vitro expression efficiencies of the two DNA vaccines were determined by Western blotting and their immunogenicities were compared by i.m. immunizing female BALB/c mice. After immunization, mice splenocytes were isolated sterilely and IFN-γ based enzyme linked immunospot assay (ELISPOT) was employed to read out the specific T cell immunity.

RESULTSThe constructed DNA vaccines were validated by restriction enzyme digestion and DNA sequencing. Western blotting result showed both of the two DNA vaccines could be expressed at appreciable levels in vitro. Under the stimulation of Consensus B Pol peptide pools, specific T cell frequency elicited by pSVAE-Pol was (636±178) SFCs/10(6) splenocytes; specific T cell frequency elicited by pSVCN-Pol was (468±265)SFCs/10(6) splenocytes (P=0.412). Under the stimulation of HIV-1 AE2f Pol peptide pools, specific T cell frequency elicited by pSVAE-Pol was (1378±611) SFCs/10(6) splenocytes; specific T cell frequency elicited by pSVCN-Pol was (713±61) SFCs/10(6) splenocytes (P=0.134). Further analysis suggested pSVAE-Pol induced specific T cell responses mainly focused on Pol 1 peptide pool, while, in addition to induce Pol 1 specific T cell responses, pSVCN-Pol could also elicit T cell responses against consensus B Pol 2 peptide pool.

CONCLUSIONAlthough pSVAE-Pol was more immunogenic, pSVCN-Pol could induce T cell responses against broader epitope spectrum. Rational vaccine design may need combine them together.

AIDS Vaccines ; genetics ; immunology ; Animals ; Female ; Genes, pol ; immunology ; HIV-1 ; genetics ; immunology ; Immunity, Cellular ; Immunization ; Mice ; Mice, Inbred BALB C ; T-Lymphocytes ; immunology ; Vaccines, DNA ; genetics ; immunology