OBJECTIVETo develop a rapid assay for simultaneous detection of HIV p24 antigen (Ag) and anti-HIV antibody (Ab).

METHODSHIV-1 gp41 antigen and HIV-2 gp36 antigen were expressed by recombinant baculovirus insect system and purified by immunochromatography. p24 monoclonal antibody (mAb) was obtained from p24 hybridoma cell line. Purified antigen and mAb were dot blotted to nitrocellular membrane; 20 nm colloidal gold-anti-human IgG ab and p24 ab complex were used for this test. Previously detected 39 sera specimens were tested in this study to compare with the result of HIV test with commercial HIV test kit.

RESULTS20 mg/L purified gp41 Ag and gp36 Ag were obtained from recombinant baculovirus-insect cell system; 1.5 mg/L p24 mAb was obtained from p24 mAb hybridoma cell line. Compared the test result of 39 sera with commercial HIV test kits, consistency rate was 100%.

CONCLUSIONSThe rapid assay for simultaneous detection of HIV p24 antigen and anti-HIV antibody provides a simple, sensitive and reliable test for HIV diagnosis.

AIDS Serodiagnosis ; Gene Products, env ; biosynthesis ; isolation & purification ; HIV Antibodies ; blood ; HIV Antigens ; biosynthesis ; isolation & purification ; HIV Core Protein p24 ; blood ; HIV Envelope Protein gp41 ; biosynthesis ; isolation & purification ; HIV Infections ; diagnosis ; HIV-1 ; immunology ; HIV-2 ; immunology ; Humans ; Reagent Kits, Diagnostic ; standards ; env Gene Products, Human Immunodeficiency Virus

OBJECTIVETo investigate the subtype distribution and the prevalence of sequence characteristics of HIV-1 strains in Beijing residents during 2006 and to analyze the relationship between distribution of HIV-1 subtypes and transmission routines.

METHODSBlood samples from 32 new confirmed HIV-1 infected individuals from Beijing residents in 2006 and separated plasma specimens were collected. RNAs were extracted and the gag and env gene were amplified by RT-PCR and nest-PCR. PCR products were sequenced directly and phylogenetic analyses of gag and env gene were performed using the MEGA2 software.

RESULTSAmong 32 HIV-1 plasma samples, 22 gag and 4 env gene fragments were amplified and analyzed. Five HIV-1 subtypes or circulating recombinant forms(CRFs) of HIV-1 including Thai B (2 strains), B (9 strains), C (2 strains), CRF07_BC (5 strains), CRF01 AE (4 strains) were identified being circulated in Beijing. The gene divergences of gag gene inside the subtypes were 6.6%, 4.3%, 6.8%, 4.9% and 3.0% in subtype B, Thai B, C, CRF01_AE and CRF07_BC respectively. Subtypes B were predominant in Beijing, accounted for 40.9% among 22 samples.

CONCLUSIONFive HIV-1 subtypes were identified in Beijing and the surveillance of HIV-1 gene variation should be paid more attention to.

China ; HIV-1 ; classification ; genetics ; Humans ; Polymerase Chain Reaction ; Reverse Transcriptase Polymerase Chain Reaction ; env Gene Products, Human Immunodeficiency Virus ; genetics ; gag Gene Products, Human Immunodeficiency Virus ; genetics

The 109 whole blood samples were collected from HIV-1 infected former blood donors in Henan and Shanxi. The RNA templates were extracted from plasma and used for the full gag gene amplification and sequencing. The sequences were divided into 3 groups according to sampling year. The Entropy software was used to identify the amino acids with composition difference among different groups of amino acid sequences. The results showed that there existed 8 and 13 amino acid sites with the statistical significance difference, respectively, in sequences in year 2004 and 2005, compared to those in 2002. Among them, there existed 5 amino acid sites in two groups. Of 16 amino acid sites, the increasing polymorphism and the decreasing polymorphism along the sampling year were observed in 10 and 6 amino acid sites respectively. Of 10 sites with increased polymorphism, 8 sites were located in the CTL epitopes recognized and presented by the main HLA alleles existed in Chinese population. The 6 sites with decreasing polymorphism all existed in main domains of Gag proteins.
Blood Donors ; China ; epidemiology ; Genetic Variation ; HIV-1 ; genetics ; Humans ; Polymorphism, Genetic ; gag Gene Products, Human Immunodeficiency Virus ; genetics

OBJECTIVETo understand the evolution of HIV-1 CRF07_BC envelope, we performed a longitudinal study on two patients during their early HIV-1 infection.

METHODSRNA was extracted from the plasma of the individuals and the C2-C5 fragments of the gp120 gene of HIV-1 were amplified by RT-PCR. Purified DNA segments were inserted into T easy vector and transformed into E. coli Top 10 competent cells. Positive clones were identified by blue-white screening, confirmed by PCR and sequenced by ABI 3700.

RESULTSThe samples were collected from the patients every 6 months from seroconversion time. The genetic diversity and divergence in env gene showed consistent increases over time. Our sequence analysis also revealed obvious non-synonymous change in env C1, C3 and V4 regions among these samples.

CONCLUSIONThe results support the concept that the consistent pattern of viral evolution existed during early phase of HIV-1 infection. C1, C3 and V4 region of env gene may be mainly immunological target during AIDS progression.

Adult ; Genetic Variation ; HIV Infections ; virology ; HIV-1 ; genetics ; isolation & purification ; Humans ; Longitudinal Studies ; Male ; env Gene Products, Human Immunodeficiency Virus ; genetics

OBJECTIVETo study the characteristics of the variation of antigen epitopes and quasispecies group in the HIV-1 viruses in Henan province in China.

METHODSThe region of the p17-p24 of the HIV-1 gag gene was amplified by nested polymerse chain reaction (PCR), purified products were cloned into the vector, the obtained were analyzed by MEGA soft wares.

RESULTSB' subtype strains were predominant in Henan province, the mutations in antigen epitypes of the p17 region of the gag gene focus on E62g (55.8%), Y79f (48.9%), T84V (48.9), I44V (44.2%), the p24 region had not found the distinct mutation.

CONCLUSIONBoth the PCR sequences and clone strains sequences demonstrated that four antigen epitopes mutations in p17 region of the HIV B' subtype gag gene, and the region of p24 was more conservative, which was suitable for development of the epitope vaccien.

Antigenic Variation ; China ; Epitopes ; genetics ; HIV Infections ; virology ; HIV-1 ; classification ; genetics ; isolation & purification ; Humans ; Molecular Sequence Data ; Mutation ; Phylogeny ; gag Gene Products, Human Immunodeficiency Virus ; genetics

OBJECTIVETo study the genetic diversity of HIV-1 nef genes from a patient with AIDS dementia complex(ADC) , so as to research the amino acid variability and the pathogenesis of ADC.

METHODSThe nef gene was amplified with PCR from genomic DNA which was extracted from spleen and different brain tissues(basal ganglia, frontal gray matter, meninges, temporal lobe)of a patient who died of ADC. PCR products were cloned into the pMD19-T vector, after transformation and selection by ampicillin and blue/white spotting. Five of positive clones were sequenced and confirmed with BLAST. HIV-1 nef sequences were processed with BioEdit and MEGA4 to do Neighbor-Joining tree, p-Distances, and values of ds/dn.

RESULTSThe samples were all identified as HIV-1 B and genetic variation exists in HIV-1 nef gene isolated from different tissues compared with HXB2. In addition,part of the changes were different between periphery and brain.

CONCLUSIONVariations exist in the HIV-1 nef gene extracted from the ADC patient and the variations from peripheral and central nerve tissues were different,these variations may change the function of Nef,and it needs more research.

AIDS Dementia Complex ; virology ; Adult ; DNA, Viral ; genetics ; Genetic Variation ; HIV Infections ; virology ; HIV-1 ; genetics ; Humans ; Male ; nef Gene Products, Human Immunodeficiency Virus ; genetics

OBJECTIVETo describe and evaluate a double-antigen sandwich ELISA for detecting human immunodeficiency virus type 1/2 (HIV-1/2) specific antibodies.

METHODSThe peptides gp41.1(sp1), gp41.2(sp2), gp120(sp3) and p24(sp4) of HIV-1 and gp36(sp5) of HIV-2 were artificially synthesized. Then sp1, sp3, sp4 and sp5 were used as coating antigens; sp1, sp2, sp4 and sp5 labeled with HRP were used as conjugates in this sandwich ELISA.

RESULTSThe specificity and sensitivity of the assay were both 100% in detecting anti-HIV of 40 control sera of the second generation panel, higher than indirect ELISA (specificity 90% and sensitivity, 65%, respectively). All of 210 sera from individuals with other diseases were negative for anti-HIV. The consistency rate was 100% when our sandwich ELISA and Abbott HIVAB were used to detect anti-HIV in 90 healthy blood donors and 88 HIV infected individuals.

CONCLUSIONSThe results showed that this sandwich ELISA for detection of anti-HIV is specific, sensitive and convenient, and it is suitable for screening blood donors and detecting HIV infection.

Enzyme-Linked Immunosorbent Assay ; methods ; HIV Antibodies ; blood ; HIV Infections ; blood ; virology ; HIV-1 ; immunology ; HIV-2 ; immunology ; Humans

We compared the performance of three screening kits for the detection of anti-HIV in 187 samples; Enzygnost Anti-HIV 1/2 Plus, ABBOTT TESTPACK HIV- 1/HIV-2 & SERODIA. HIV- 1/2. Four samples, 3 serums and 1 CSF, from 2 patients were repeatedly reactive in all three screening kits and 2 serum specimens were confirmed positive(HIV-1) by the western blot assay. The sensitivity and specificity of all three screening kits were 100% and 98.9%, respectively. In Korea, the cause of AIDS is mostly HIV-1 and the prevalence is very low. So, all three screening kits were useful for the detection of anti-HIV from patients and blood donors. But the use of screening kit for the detection of anti-HIV-1, anti-HIV-2 and anti-HIV-I subtype O will be needed for the decrement of false negative rate because HIV infection has been increased, especially, HIV-2 infection and pediatric AIDS patient by vertical transmission were also reported, currently.
Antibodies* ; Blood Donors ; Blotting, Western ; HIV Infections ; HIV* ; HIV-1 ; HIV-2 ; Humans ; Korea ; Mass Screening* ; Prevalence ; Sensitivity and Specificity

Apolipoprotein B mRNA-editing enzyme catalytic polypeptide 3 protein G (APOBEC3G) is part of the innate immune system of host cells and has cytidine deaminase activity. It specifically incorporates into the virion during HIV-1 replication. The incorporation of APOBEC3G needs its interaction with HIV-1 Gag. In the HIV-1 reverse transcription process, APOBEC3G deaminates dC to dU in the first minus strand cDNA, and then induces extensive hypermutation in the viral genome. Besides deamination, APOBEC3G also inhibits HIV-1 by some kinds of non-deamination mechanisms which need to be further elucidated. HIV-1 Vif counteracts the activity of APOBEC3G by an ubiquitin-proteasome-mediated degradation of APOBEC3G. As a broad spectrum inhibitor of viruses, APOBEC3G also inhibits various retroviruses, retrotransposons and other viruses like HBV. Upregulating the expression of APOBEC3G or blocking the Vif-mediated degradation of APOBEC3G might be novel strategies to treat HIV-1 infection in the future.
APOBEC-3G Deaminase ; Amino Acid Substitution ; Anti-HIV Agents ; metabolism ; Cytidine Deaminase ; genetics ; metabolism ; Gene Expression ; HIV Infections ; metabolism ; HIV-1 ; genetics ; physiology ; Humans ; Virus Replication ; vif Gene Products, Human Immunodeficiency Virus ; genetics ; metabolism

No abstract available.
eIF-2 Kinase* ; HIV-1* ; Phosphorylation