[Molecular investigation of a possible case of HIV transmission after a blood transfusion].

OBJECTIVE: A molecular technique based on quasispecies analysis for tracing postexposure HIV transmission was applied in an investigation of a possible case of HIV transmission after blood transfusion. METHODS: Sixteen plasma specimens were collected from 3 HIV infections (T1-T3) involved in a possible HIV transmission chain and 13 HIV/AIDS (C1-C13) controls. The RNAs were extracted and then amplified by RT-PCR, the PCR products were cloned and sequenced.BioEdit 6.0.7 and MEGA 4.0 software were used to analyze gene sequences, calculate gene dispersion ratio and construct phylogenetic tree. RESULTS: The sequences of 13 specimens were successfully obtained.The HIV strains from T1, T2 and T3 were CRF07_BC recombinants, those from 5 out of the 6 controls lived in the same city with T2 and T3 were CRF07_BC recombinants as well, while those from 4 controls living in the same city with T1 were CRF01_AE recombinants. Compared with the clone sequences from T1, the mean gene dispersion ratio of T2 was the least (2.0%), followed by C12 (2.8%) , T3 (2.9%) and others. The phylogenetic tree showed that all clones from T1, T2, T3 and C12 might cluster together,and implied that the direction of HIV transmission was from T3 to T2, and then to T1. CONCLUSION: The results support the possible epidemiological clue that HIV was transmitted from T3 to T2, and then to T1, indicating that molecular epidemiological investigation could provide more direct evidence for tracing postexposure HIV transmission.

[Comparison of three HIV antibody confirmatory assay kits in confirming early HIV infection].

OBJECTIVE: This study was to compare the performance of three HIV antibody confirmatory assay kits in confirming early HIV infection. METHODS: Five HIV antibody-positive plasma specimens were ten-fold serially diluted and then detected by ELISA. The above diluted specimens were detected with the following three HIV antibody confirmatory assay kits to analyze their sensitivity, including Wantai-RIBA (Recombinant immunoblot assay, Beijing Wantai Biological Pharmacy, China), MP-WB (HIV Blot 2.2 WB, MP Biomedicals Asia Pacific Pte. Ltd., Singapore) and INNO-LIA (INNO-LIA(TM) HIV I/II Score, Innogenetics N.V., Belgium), respectively. These kits were further used to detect 48 ELISA-reactive specimens from 11 sets of HIV seroconversion specimens (a total of 48 samples) which were previously detected as HIV antibody-positive by ELISA. RESULTS: When 5 samples were diluted to 100 fold, Wantai-RIBA still can detect them positive. Among the 48 HIV antibody-positive specimens detected with ELISA, the confirmation positive rate for Wantai-RIBA, MP-WB and INNO-LIA were 97.92% (47/48), 81.25% (39/48) and 91.67% (44/48), respectively. There was statistically significant difference between the confirmatory results of Wantai-RIBA and MP-WB (χ(2) = 6.13, P < 0.05), as well as between those of INNO-LIA and MP-WB (χ(2) = 5.48, P < 0.05); however, there was no statistically significant difference between those of Wantai-RIBA and INNO-LIA (χ(2) = 1.33, P > 0.05). For other six HIV seroconversion panels containing indeterminate specimens, the average seroconversion period of time for Wantai-RIBA, MP-WB and INNO-LIA were 0.7, 13.3 and 3.7 days, respectively. CONCLUSION: Compared with MP-WB, Wantai-RIBA and INNO-LIA could reduce the window period to confirm early HIV infection.

Human immunodeficiency virus-1 genotypic drug resistance among volunteer blood donors in Yunnan, China.

BACKGROUND: Drug resistance profiles of human immunodeficiency virus-1 (HIV-1) in treatment-naïve infections have been reported in developed countries. However, little is known in developing countries, including China, especially in treatment-naïve volunteer blood donors. STUDY DESIGN AND METHODS: Fifty-two HIV-1-positive samples of blood donors were collected from 2005 to 2006 in Yunnan, China. Recent and long-term infections were distinguished by the HIV-1 subtypes B, E, and D immunoglobulin G-capture enzyme immunoassay assay. The nucleotide sequences of pol genes were amplified and sequenced. Phylogenetic tree and drug resistance analyses were performed. RESULTS: Of 49 samples successfully analyzed, circulating strains were circulating recombinant form (CRF)08_BC (51.0%), CRF07_BC (24.5%), CRF01_AE (20.4%), and B (4.1%). No protease inhibitors (PI) major drug resistance mutation (DRM) was detected. Six samples (12.2%) displayed seven minor PI DRMs. Nine samples (18.4%) displayed 10 nucleoside reverse transcriptase inhibitor DRMs, and DRMs to nonnucleoside reverse transcriptase inhibitors were present in one sample (2.0%). There was only one sample of the 49 (2.0%) in which the DRMs were of sufficient magnitude to result in a clinical change to drug susceptibility, but even in this sample, the clinical effect of these DRMs was predicted to be low. Significant differences were not observed between the long-term and recent infected population. Differences in DRMs were not observed between peripheral blood mononuclear cells and plasma within an individual. CONCLUSIONS: CRF_BC was the dominant subtype circulating in HIV-1-infected donors in Yunnan. Prevalence of genotypic drug resistances among donors in Yunnan was low in this study. Surveillance on HIV-1 infections among blood donors should be continued in China.

[Study on the influence of biological characteristics to HIV/HCV co-infection among HIV infected illegal blood donors in China].

OBJECTIVE: To study the influence of biological characteristics on HCV/HIV co-infection, including HIV-1 sequence distance, HIV-1 viral load and CD4+ count. METHODS: HIV-1 sequence distance was calculated by Clustal W and Phylip software while HIV-1 viral load being tested by NASBA and CD4+ count was tested using Epics XL of Coulter. Significance was determined by t-test using SPSS 12.0. RESULTS: The mean HIV-1 genetic distances were 7.95% and 15.73% (P < 0.001) between those with HCV co-infection and those without. Their mean HIV-1 viral loads were 4.61 and 4.45 (P = 0.522) and their mean CD4I T counts were 308 and 251 (P = 0.161), respectively. CONCLUSION: Data showed that in the study group, the HIV/HCV co-infection had an influence on the HIV sequence distance, but did not have major impact on HIV-1 viral load and their mean CD4+ T count.

[Comparison of NucliSens HIV-1 QT and Amplicor HIV-1 monitor 1.5 in detecting HIV-1 viral load].

OBJECTIVE: To compare the results of detecting HIV-1 load by using NucliSens HIV-1 QT and Amplicor HIV-1 monitor 1.5 assays. METHODS: Eighty-two clinical samples were collected and HIV viral load was determined with the above-mentioned two methods. RESULTS: The number of samples in which values obtained by NucliSens HIV-1 QT and Amplicor HIV-1 monitor 1.5 differed by <0.5 log10 RNA copies/ml and in which the viral load was undetectable accounted for 88.9 percent of the measures. The correlation coefficient between the two methods was 0.956 in 56 samples of Deltalog10 VL<0.5. CONCLUSION: The results of HIV-1 viral load determination with the two methods are highly comparable.

[Study on the use of dried blood spot in sequencing and subtyping HIV-1 DNA genome].

OBJECTIVE: To use dried blood spot (DBS) in studying the sequence and subtype analysis of HIV-1 genome. METHODS: 2 ml whole blood containing EDTA anticoagulant from 20 HIV-1 infected patients were collected, then 80 microl blood was used to propare DBS. QIAamp Blood Mini kit and 10% Chelex100 resin extracted DNA genome from whole blood and DBS as well as nested PCR amplified specifically HIV-1 Gp41 region from the two kinds of DNA extraction. Software MAGE 3.0 was used to study the sequence and subtype of the PCR products. RESULTS: Eligible 18 paired samples were analysed to show that 16 of them belonged to C54A, 97CNGX-7F, 98CN006 subtypes. The other two samples might belong to B. CN._. RL42 and B. US. 90WEAU160. CONCLUSION: Data showed that there were parallel results between the whole blood and DBS samples including subtype analysis, position of mutation and types of amino acid sequencing. Since DBS itself facilitated the collection, transportation and storage, it could be used as a measure to collect blood sample in resource limited area and to develop molecular epidemiologic research as well as early diagnosis on infant exposed to HIV.

[Heterogeneity of HIV strains isolated from different tissues of 3 AIDS patients].

BACKGROUND: To reveal the characteristics of genotype and phenotype of HIV strains in blood and some tissues of AIDS patients. METHODS: The virus was isolated from peripheral blood mononuclear cell (PBMC),cerebrospinal fluid (CSF)and lymph nodes of 3 AIDS patients by coculture with PBMC stimulated by PHA for 72 hours from uninfected donor. The cytopathic effect of the HIV isolates was determined in cultured MT2 cell line. The env gene sequences form proviral DNA were analyzed by GCG software. RESULTS: In one patient,there were differences between the strains from blood and different tissues both in genotype and phenotype. The biological phenotypes of two strains from CSF were non syncytium (NSI) type, their env sequences were similar to standard CNS tropic strain (SF162). CONCLUSIONS: The viral heterogeneity exists in different body compartments within an infected individual. The neurotropic isolate which is similar to international standard strain exists in some AIDS patients in China.