Teixeira Daniela, Munerato Patricia, Komninakis Shirley Cavalcante Vasconcelos, Fusuma Erika Etsuko, Janini Luiz Mario, Sucupira Maria Cecilia Araripe, Diaz Ricardo Sobhie
Retrovirology Laboratory, Paulista School of Medicine, Federal University of São Paulo, São Paulo, Brazil.
AIDS Res Hum Retroviruses. 2010 Sep;26(9):981-90. doi: 10.1089/aid.2010.0023.
We sought to determine the frequency and profile of HIV-1 BF recombinants in vitro and in vivo. Laboratory HIV-1 strains from subtypes B and F were cocultured and evaluated. Clinical samples from the city of Santos, Brazil, where the first HIV-1 B/F circulating recombinant forms (CRF) were described, were also assessed. Five real-time PCR assays were developed to equally amplify subtypes B and F, and subtype-specific probes were developed and optimized. To validate the PCR systems, clinical samples from Santos were sequenced and phylogenetically analyzed. The real-time PCR assays were performed on these samples and on the supernatant of an in vitro competition assay to assess emergent recombinant strains. Out of 157 clinical samples, 62.1% were defined as subtype B, 3.0% were subtype F, 16.7% presented the CRF28_BF profile, and 13.6% of the samples presented the CRF29_BF profile. The specificity and sensitivity in the discrimination assay for this sample panel were 93% and 92%, respectively. The HIV that emerged from the coinfected cell culture closely resembled the CRF28_BF profile. The first-described CRFs are still fixed in this geographic region of Brazil, and the in vitro emerging strains detected by real-time PCR suggest that in addition to the shaping of recombinant strains by immune selection, viral structures may also play an important role in emerging CRFs.
我们试图确定HIV-1 BF重组体在体外和体内的频率及特征。对来自B亚型和F亚型的实验室HIV-1毒株进行共培养并评估。还对巴西桑托斯市的临床样本进行了评估，该市曾发现首例HIV-1 B/F循环重组形式（CRF）。开发了5种实时PCR检测方法以同等程度地扩增B亚型和F亚型，并开发和优化了亚型特异性探针。为验证PCR系统，对来自桑托斯的临床样本进行测序和系统发育分析。对这些样本以及体外竞争试验的上清液进行实时PCR检测，以评估新出现的重组毒株。在157份临床样本中，62.1%被确定为B亚型，3.0%为F亚型，16.7%呈现CRF28_BF特征，13.6%的样本呈现CRF29_BF特征。该样本组鉴别试验的特异性和灵敏度分别为93%和92%。共感染细胞培养中出现的HIV与CRF28_BF特征非常相似。首次描述的CRF在巴西的这个地理区域仍然固定存在，通过实时PCR检测到的体外新出现毒株表明，除了免疫选择对重组毒株的塑造作用外，病毒结构可能在新出现的CRF中也发挥着重要作用。
