The Detection of in Vivo and in Vitro HIV Type 1 B/F Profiles in Brazil Using a Real-Time PCR Assay for Five HIV Type 1 Genomic Regions

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.Department of STD/AIDS of the Ministry of HealthFundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Universidade Federal de São Paulo, Paulista Sch Med, Retrovirol Lab, BR-04039032 São Paulo, BrazilUniversidade Federal de São Paulo, Paulista Sch Med, Retrovirol Lab, BR-04039032 São Paulo, BrazilFAPESP: 04/15856-9FAPESP: 06/52722-6Web of Scienc

Postoperative Ileus: Pathophysiology, Prevention and Treatment

Background: Continuous long-term treatment is recommended to reduce the hepatitis B virus (HBV) viral load. However, as a consequence, resistance mutations can emerge and be transmitted to other individuals. the polymerase (POL) gene overlaps the surface (S) gene. Thus, during treatment, mutations in the POL gene may lead to changes in hepatitis B surface antigen (HBsAg). the purpose of this study was to evaluate the frequency of lamivudine and vaccine escape mutations in HBsAg-positive blood donors from the city of Santos and in untreated HBV mono-infected patients from the city of São Paulo, Brazil.Methods: HBV DNA was extracted from 80 serum samples, of which 61 were from volunteer blood donors and 19 were from untreated HBV patients. A fragment of the POL/S genes containing 593 base pairs was amplified using nested PCR. Thirty four were PCR-positive and sequencing was performed using an ABI Prism 3130 Genetic Analyzer. Alignments and mutation mapping were performed using BioEdit software.Results: HBV DNA from 21 blood donors and 13 untreated patient samples were characterized using nucleotide sequencing PCR products from the POL/S genes. We were able to detect one sample with the resistance mutation to lamivudine rtM204V + rtL180M (2.94%), which was found in a volunteer blood donor that has never used antiviral drugs. the other samples showed only compensatory mutations, such as rtL80F (5.88%), rtL80V (2.94%), rtL82V + rtV207L (2.94%), rtT128P (5.88%), rtT128N/S (2.94%) and rtS219A (5.88%). We found modifications in the S gene in 14 of the 34 samples (41.16%). the mutations detected were as follows: sM133L + sI195T (2.94%), sI195M (2.94%), sP120T (2.94%), sY100S/F (2.94%), sY100C (17.64%), sI/T126P + sQ129P (2.94%), sM198I + sF183C (2.94%) and sS210R (5.88%).Conclusions: Our results suggest the transmission of lamivudine-resistant forms. Thus, the evaluation of HBV-infected subjects for lamivudine resistance would improve treatment regime. Moreover, the mutations in the S gene may impair HBsAg antigenicity and contribute to HBsAg failure detection and vaccine escape

Wuhan large pig roundworm virus identified in human feces in Brazil

We report here the complete genome sequence of a bipartite virus, herein denoted WLPRV/human/BRA/TO-34/201, from a sample collected in 2015 from a two-year-old child in Brazil presenting acute gastroenteritis. The virus has 98-99% identity (segments 2 and 1, respectively) with the Wuhan large pig roundworm virus (unclassified RNA virus) that was recently discovered in the stomachs of pigs from China. This is the first report of a Wuhan large pig roundworm virus detected in human specimens, and the second genome described worldwide. However, the generation of more sequence data and further functional studies are required to fully understand the ecology, epidemiology, and evolution of this new unclassified virus.FAPESPCNPqAdolfo Lutz Inst, Virol Ctr, Enter Dis Lab, Av Dr Arnaldo 355, BR-01246902 Sao Paulo, SP, BrazilFed Univ Para, Inst Biol Sci, Belem, Para, BrazilFac Med ABC, Postgrad Program Hlth Sci, Santo Andre, BrazilUniv Fed Sao Paulo, Retrovirol Lab, Sao Paulo, SP, BrazilUniv Sao Paulo, Fac Med, LIM 46, Sao Paulo, BrazilFed Univ Tocantins, Palmas, Tocantins, BrazilPubl Hlth Lab Tocantins State LACEN TO, Palmas, BrazilUniv Sao Paulo, Inst Trop Med, Av Dr Eneas de Carvalho Aguiar 470, BR-05403000 Sao Paulo, SP, BrazilBlood Syst Res Inst, San Francisco, CA USAUniv Calif San Francisco, Dept Lab Med, San Francisco, CA 94143 USAUniv Fed Sao Paulo, Retrovirol Lab, Sao Paulo, SP, BrazilFAPESP: 2015/12944-9FAPESP: 2017/00021-9FAPESP: 2016/01735-2CNPq: 400354/2016-0Web of Scienc

A Novel Highly Divergent Strain of Cell Fusing Agent Virus (CFAV) in Mosquitoes from the Brazilian Amazon Region

Classical insect-specific flaviviruses (cISFs) have been widely detected in different countries in the last decades. Here, we characterize the near full-length genomes of two cISFs detected in mosquitoes collected in the city of Macapá, state of Amapá, Amazon region of Brazil. A total of 105 pools of female mosquitos were analyzed by next-generation sequencing (NGS). Comparative genomics and phylogenetic analysis identified three strains of cell fusing agent virus (CFAV) and two of Culex flavivirus (CxFV). All sequences were obtained from pools of Culex sp., except for one sequence of CFAV detected in a pool of Aedes aegypti. Both CxFV strains are phylogenetically related to a strain isolated in 2012 in the Southeast region of Brazil. The CFAV strains are the first of this species to be identified in Brazil and one of them is highly divergent from other strains of CFAV that have been detected worldwide. In conclusion, CFAV and CxFV, circulate in mosquitoes in Brazil. One strain of CFAV is highly divergent from others previously described, suggesting that a novel strain of CFAV is present in this region