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目的了解广东省艾滋病病毒(HIV)主要流行亚型及V3环氨基酸的变异特点,并分析病毒株利用辅助受体的情况。方法收集2013-2014年期间,在广州市第八人民医院住院的艾滋病(AIDS)病人的血浆256份,巢式聚合酶链反应(Nested-PCR)方法扩增env基因C2-C3(HBX2:6972~7365)区并直接测序,用Geen tool软件将基因序列翻译为氨基酸序列进行比对和分析。结果共有214例(83.6%)扩增成功,其中143例(66.8%)为CRF01_AE;45例(21.0%)为B/C重组型;26例(12.1%)为CRF01_AE/B亚型。V3环顶端四肽基序,73.4%(157/214)为GPGQ,11.7%(25/214)为GPGR,8.9%(19/214)为GPGK,2.3%(5/214)为GQGQ,其他为(GPGG、GLGH、GPGS、GPGH)3.7%(8/214)。使用国际上普遍应用的4种软件(PSSMx4r5、SAAC+BLAS、PSSMsisi、Geno2pheno)分别对V3环顶端四肽基序利用辅助受体的情况进行评估。综合评估结果显示,132例(61.7%)利用CCR5,67例(31.3%)利用CXCR4。在15例(7.0%)无辅助受体评估结果的病例中,14例为CRF01_AE,1例为CRF01_AE/B。研究还发现,71.1%(32/45)B/C重组型的V3四肽基序单一使用PSSMsisi软件的评估结果,与4种软件综合评估结果的一致率为100%。结论目前广东省HIV-1的主要流行株仍是CRF01_AE,其V3环氨基酸高度变异,顶端四肽基序主要是GPGQ。对毒株利用辅助受体的情况,宜采取国际普遍使用的4种软件进行综合分析。
Objective To understand the characteristics of the major epidemic subtypes of HIV in Guangdong Province and the variation of amino acids in V3 loop and to analyze the utilization of co-receptors by virus strains. Methods Totally 256 plasma samples were collected from patients with AIDS in Guangzhou Eighth People’s Hospital during 2013-2014. The env gene C2-C3 (HBX2: 6972) was amplified by nested polymerase chain reaction (Nested-PCR) ~ 7365) region and direct sequencing, using Geen tool software to translate the gene sequence into amino acid sequence for comparison and analysis. Results A total of 214 cases (83.6%) were successfully expanded, 143 (66.8%) were CRF01_AE, 45 (21.0%) were B / C and 26 (12.1%) were CRF01_AE / B subtypes. V3 loop apical tetrapeptide motif, GPGQ was 73.4% (157/214), GPGR was 11.7% (25/214), GPGK was 8.9% (19/214), GQGQ was 2.3% (5/214), others were (GPGG, GLGH, GPGS, GPGH) 3.7% (8/214). The use of co-receptors for the apical tetrapeptide motif of the V3 loop was evaluated using four softwares commonly used in the world (PSSMx4r5, SAAC + BLAS, PSSMsisi, Geno2pheno). The comprehensive assessment showed that 132 patients (61.7%) were using CCR5 and 67 (31.3%) were using CXCR4. Of the 15 patients (7.0%) who did not have a co-receptor assessment, 14 were CRF01_AE and 1 was CRF01_AE / B. The study also found that 71.1% (32/45) of the B / C recombinant V3 tetrapeptide motifs were evaluated using the PSSMsisi software alone and were 100% identical to the four softwares. Conclusions At present, the major epidemic strain of HIV-1 in Guangdong Province is still CRF01_AE. The V3 loop amino acid is highly variable and the top tetrapeptide motif is mainly GPGQ. The use of co-receptors on the strains, the appropriate use of internationally accepted 4 kinds of software for a comprehensive analysis.