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 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

To study the CTL antigen epitopes and drug resistance mutations of HIV-1 gag and pol genes through analyzing gag and pol gene sequences. The HIV-1 gag and pol gene fragments were amplified using nested polymerase chain reaction. A total of 23 PCR sequences, 449 cloned gag sequences and 402 cloned pol sequences were obtained. Sequence analyses showed the 23 samples were subtype B or B'. A total of 4 in 8 CTL antigen epitopes appeared 8 mutations in consensus sequence of subtype B and B'. There were no mutations found in the PCR sequences, whereas a few mutations were found in clone sequences (9.80%) in 5 antigen epitopes in p24 region. Eighteen PIs-related mutations and 24 RTIs-related mutations were found in PCR sequences and clone sequences in pol gene region, in which 17 (94.44%) PIs-related mutations and 15 (62.50%) RTIs-related mutations were found only in the clone sequences, respectively. The results showed that the prevalence of HIV-1 drug resistance strains in this study was at a higher level (17.39%), suggesting that some samples were resistant.to existing antiviral drugs.
Antigens, Viral ; immunology ; DNA Mutational Analysis ; Drug Resistance, Viral ; genetics ; Epitopes ; immunology ; HIV-1 ; classification ; drug effects ; genetics ; immunology ; Human Immunodeficiency Virus Proteins ; genetics ; Mutation ; Phylogeny ; T-Lymphocytes, Cytotoxic ; immunology ; gag Gene Products, Human Immunodeficiency Virus ; genetics ; pol Gene Products, Human Immunodeficiency Virus ; genetics

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

To investigate the subtype distribution of human immunodeficiency virus-1(HIV-1) infection among men who have sex with men (MSM) in Zhengzhou, Henan Province, forty blood samples were collected from HIV-1 carriers, who acknowledged to have sex with men. The complete gag gene was amplified by RT-PCR and nested-PCR and sequenced. All sequences were edited by BioEdit and subtyped by genotyping software. Phylogenetic analysis of gag gene were then performed using the MEGA 3.1 software, the gene distances were calculated by Distance program. There were three different HIV-1 subtypes including B, CRF01-AE and CRF07-BC present among twenty four MSMs in Zhengzhou. Genotyping results showed that 33.33% (8/24) were B, 41.67% (10/24) were CRF01-AE and 25% (6/24) were CRF07-BC, and subtype CRF01-AE had become the most prevalent HIV-1 subtype in Zhengzhou, Henan province. In conclusion, recombinant HIV-1 strains are circulating in Henan province and the epidemiology is complicated.
Adult ; China ; HIV-1 ; classification ; genetics ; isolation & purification ; Homosexuality, Male ; Humans ; Male ; Middle Aged ; Phylogeny ; Sequence Analysis, DNA ; Young Adult ; gag Gene Products, Human Immunodeficiency Virus ; genetics