Homogenous HIV-1 subtype B from the Brazilian Amazon with infrequent diverse BF1 recombinants, subtypes F1 and C among blood donors.

In the last decade a growing HIV/AIDS epidemic with increased incidence and AIDS-related mortality has been reported in Northern Brazil from which molecular data are scarce. Also, apparently healthy, adult blood donors, recently diagnosed with HIV-1 represent important sentinel populations for molecular studies. This cross-sectional study describes HIV-1 subtypes in blood donors from three reference public blood centers located in three States in Northern Brazil. HIV-1 pol sequencing (protease/PR, reverse transcriptase/RT) was performed on plasma samples of HIV-1 positive donors from HEMOAM, Manaus, Amazonas (n = 198), HEMERON, Porto Velho, Rondônia (n = 20) and HEMORAIMA, Boa Vista, Roraima (n = 9) collected from 2011-2017. HIV-1 subtypes were identified by REGA, phylogenetic inference; recombinant viruses were characterized by SIMPLOT. Young, single, males predominated, around half was first-time donors. Syphilis co-infection was detected in 17% (39 out of 227), 8% (18 out of 227) was anti-HBc positive. Subtype B represented ≥ 90% in Amazonas, Rondônia and Roraima, subtype C (3.1%) was found in Amazonas and Rondônia; subtype F1 (0.9%) and BF1 recombinants (5.3%) were only detected in Amazonas. Subtype B sequences from Amazonas (n = 179), Rondônia (n = 18) and Roraima (n = 9) were combined with viral strains representative of the BPANDEMIC (n = 300) and BCARIBBEAN/BCAR (n = 200) lineages. The BPANDEMIC lineage predominated (78%) although BCAR lineages were frequent in Roraima (56%) and Amazonas (22%). Subtype C and subtype F1 sequences identified here clustered within Brazilian CBR and F1BR lineages, respectively. Twelve BF1 mosaics showed 11 different recombination profiles: six were singleton unique-recombinant-forms/URFs, one displays a CRF28/29_BF-like recombinant pattern and the remaining four BF1 isolates branched with other Brazilian BF1 viruses previously described and may represent putative new CRF_BF1 from Northern Brazil. Our study shows a highly homogeneous molecular pattern with prevalent subtype B, followed by BF1, and sporadic subtype C and F1 in blood donors from the Northern region. Surveillance studies are important to monitor HIV-1 diversity which can reveal patterns of viral dissemination, especially in a highly endemic, remote and geographically isolated region as Northern Brazil.

High rate of seromarkers for HIV, HBV and syphilis among blood donors using confidential unit exclusion, before and after HIV-NAT implementation at a major public blood bank in the Brazilian Amazon.

BACKGROUND: Confidential unit exclusion (CUE) was introduced in the 1980's as an additional layer to blood safety, before highly specific and sensitive nucleic acid tests (NAT) for HIV were implemented. The utility of CUE-use in settings that have implemented NAT should be evaluated over time. STUDY DESIGN, METHODS: Cross-sectional retrospective study carried out from June 2010-November 2015, at Manaus Hemocenter (HEMOAM), Amazonas, Brazil that implemented HIV-NAT in 2012. The HIV, HCV, HBV, HTLV, Chagas disease, and syphilis rates were compared among CUE and non-CUE blood donors, before and after HIV-NAT implementation. RESULTS: Among 287,588 donations, 2,154 (0.75%) were associated with CUE, mainly voluntary donations (64.2%), by repeat donors (58.4%) from young (median age = 31 years), males (84.4%), unmarried (63.1%). CUE-users compared to non-CUE donors (n = 285,434) had higher seropositivity rates to HIV (OR = 6.09, 95% CI: 3.68-10.07, p < 0.001), HBV (anti-HBc OR = 1.81 95% CI: 1.24-2.64, p = 0.004; HBsAg OR = 5.68, 95% CI: 1.78-18.07, p = 0.017), and syphilis (OR = 1.78, 95% CI: 1.05-3.04, p = 0.030). Most (97.2%) discarded blood units associated to CUE was seronegative for all pathogens. Most donations (73.4%) were tested by HIV-NAT and showed four window period donations, positive by HIV-NAT only among non-CUE donors. CONCLUSION: A high rate of transfusion transmissible infections/TTIs was observed at HEMOAM especially in CUE-users. CUE-use offered an additional layer of blood safety by its association with anti-HBc/HBsAg and syphilis that are not covered by NAT. For blood banks in highly endemic areas for HIV and TTI, as HEMOAM, the identification of at risk donors, and the orientation to be tested at proper sites remain a great challenge.

Clinical and laboratory features of HTLV-I asymptomatic carriers and patients with HTLV-I-associated myelopathy/tropical spastic paraparesis from the Brazilian Amazon.

Clinical and laboratory parameters including blood and cerebrospinal fluid (CSF) neopterin were investigated in human-T-lymphotropic-virus-type-I associated-myelopathy/tropical-spastic-paraparesis-HAM/TSP and in HTLV-I carriers. HAM/TSP (n = 11, 2 males/9 females, median age = 48 years), recently diagnosed HTLV-I carriers (n = 21, 15 females/6 males, median age = 44 years), healthy individuals (n = 20, 10 males/10 females, median age = 34.6 years) from the Brazilian Amazon (Manaus, Amazonas State) were investigated. Neopterin was measured (IBL ELISA Neopterin, Germany) in serum samples of all the participants, in CSF of 9 HAM/TSP patients as well as in 6 carriers. In HAM/TSP patients, CSF cell counts, protein and glucose were measured, the Osame's motor-disability-score/OMDS was determined, and brain/spinal cord magnetic-resonance-imaging (MRI) was performed. HAM/TSP patients had normal CSF glucose, leukocyte counts; and normal protein levels predominated. Brain-MRI showed white-matter lesions in 7 out of 11 HAM/TSP patients. OMDS varied from 2-8: 9 were able to walk, 2 were wheel-chair-users. The median serum neopterin concentration in HAM/TSP patients was 6.6 nmol/ L; min. 2.8- max. 12.5 nmol/ L); was lower in carriers (4.3 nmol/L; min. 2.7- max. 7.2 nmol/ L) as well as in healthy participants (4.7 nmol/ L; min. 2.7- max. 8.0 nmol/ L) (p < 0.05). CSF neopterin concentrations in HAM/TSP patients were higher than in serum samples, and higher compared to carriers (p < 0.05). Carriers had similar serum-CSF neopterin concentrations compared to healthy participants. Variable clinical and laboratory profiles were seen in HAM/TSP patients, however our results support the neopterin measurement as a potential biomarker of disease activity.

Clinical and laboratory features of HTLV-I asymptomatic carriers and patients with HTLV-I-associated myelopathy/tropical spastic paraparesis from the Brazilian Amazon

ABSTRACT Clinical and laboratory parameters including blood and cerebrospinal fluid (CSF) neopterin were investigated in human-T-lymphotropic-virus-type-I associated-myelopathy/tropical-spastic-paraparesis-HAM/TSP and in HTLV-I carriers. HAM/TSP (n = 11, 2 males/9 females, median age = 48 years), recently diagnosed HTLV-I carriers (n = 21, 15 females/6 males, median age = 44 years), healthy individuals (n = 20, 10 males/10 females, median age = 34.6 years) from the Brazilian Amazon (Manaus, Amazonas State) were investigated. Neopterin was measured (IBL ELISA Neopterin, Germany) in serum samples of all the participants, in CSF of 9 HAM/TSP patients as well as in 6 carriers. In HAM/TSP patients, CSF cell counts, protein and glucose were measured, the Osame’s motor-disability-score/OMDS was determined, and brain/spinal cord magnetic-resonance-imaging (MRI) was performed. HAM/TSP patients had normal CSF glucose, leukocyte counts; and normal protein levels predominated. Brain-MRI showed white-matter lesions in 7 out of 11 HAM/TSP patients. OMDS varied from 2-8: 9 were able to walk, 2 were wheel-chair-users. The median serum neopterin concentration in HAM/TSP patients was 6.6 nmol/ L; min. 2.8- max. 12.5 nmol/ L); was lower in carriers (4.3 nmol/L; min. 2.7- max. 7.2 nmol/ L) as well as in healthy participants (4.7 nmol/ L; min. 2.7- max. 8.0 nmol/ L) (p < 0.05). CSF neopterin concentrations in HAM/TSP patients were higher than in serum samples, and higher compared to carriers (p < 0.05). Carriers had similar serum-CSF neopterin concentrations compared to healthy participants. Variable clinical and laboratory profiles were seen in HAM/TSP patients, however our results support the neopterin measurement as a potential biomarker of disease activity.

Molecular epidemiology of hepatitis B and hepatitis delta viruses circulating in the Western Amazon region, North Brazil.

BACKGROUND: Hepatitis B virus (HBV) and hepatitis D virus (HDV) represent important public health problems in the Western Amazon region with reported cases of fulminant hepatitis. This cross sectional study describes HBV and HDV genotypes circulating in the Brazilian Amazon region. METHODS: HBsAg positive individuals (n = 224) were recruited in Manaus/Amazonas State (130 blood donors from the Hematology and Hemotherapy Foundation from Amazonas/HEMOAM; 60 subjects from outpatient clinic) and in Eirunepe city (n = 34) from 2003-2009. Most participants (n = 153) lived in Manaus, 63 were from 20 remote isolated municipalities, 8 lived outside Amazonas State. Genotyping was based on PCR products: HBV genotype A-F specific primers, restricted length polymorphism for HDV. HDV isolates were directly sequenced (delta antigen 405 nucleotide fragment) and phylogenetic analysis performed (MEGA; neighbor-joining, Kimura's two parameter). RESULTS: Most participants were young adult males and HBV mono-infection predominated (70.5%, 158/224). Among blood donors, outpatient subjects and individuals from Eirunepe, HBV/A prevailed followed by HBV/D and F (p > 0.05). HBV/A was more frequent in blood donors (p < 0.05). HBV-HDV coinfection rate was 8.5% in blood donors (11/130), 65.0% (39/60) in outpatient subjects and 47.0% (16/34) in individuals from Eirunepe. Compared to blood donors, coinfection was higher in outpatient subjects (65.0% versus 8.5%; RR = 5.0; CI 3.4-7.9; p < 0.0001) and in subjects from Eirunepe (47.0% versus 8.5%; RR = 5.5; CI 3.0-9.9; p < 0.0001). HBV-HDV coinfection rates were higher in patients from highly endemic remote cities. Only HDV genotype 3 was detected, HBV/F-HDV/3 predominated (20/38; 52.7%), followed by HBV/A-HDV/3 (31.6%; 12/38) and HBV/D-HDV/3 (15.8%; 6/38). CONCLUSIONS: The description of HBV and HDV genotypes circulating in the western Amazon can contribute to a better understanding of their relevance on the regional epidemics. These infections are highly endemic in the Amazon where their control is challenged by its vast territorial dimension with small, hard-to-reach municipalities dispersed into the jungle and populated by diverse ethnic groups.

Influence of hepatitis B virus (HBV) genotype on the clinical course of disease in patients coinfected with HBV and hepatitis delta virus.

OBJECTIVE: We evaluated the influence of hepatitis B virus (HBV) genotype on the course of disease in patients coinfected with HBV and hepatitis delta virus (HDV). METHODS: We evaluated HBV genotypes in 190 patients, 140 of whom had chronic HBV monoinfection and 50 of whom had chronic HBV-HDV coinfection. Real-time polymerase chain reactions for the amplification of HBV DNA and HDV RNA were developed, and we compared the patient groups with respect to HBV genotype, viral load, alanine aminotransferase (ALT) and bilirubin levels, and disease severity. RESULTS: Coinfected patients had higher ALT and bilirubin levels as well as a higher prevalence of liver cirrhosis and liver carcinoma. ALT levels were higher among individuals coinfected with HDV and HBV genotype F than among individuals infected only with HBV genotype F. Among HDV-HBV-coinfected patients, HDV load was lower among those infected with HBV genotype A than among those infected with HBV genotype D or genotype F. CONCLUSION: Liver inflammation and HDV load are influenced by HBV genotype in individuals coinfected with HBV and HDV.

Prevalência de marcadores sorológicos e moleculares do vírus da hepatite B em gestantes do Estado do Amazonas, Brasil / Prevalence of serologic and molecular markers of hepatitis B virus infectio among pregnant women in Amazonas State, Brazil

A infecção pelo vírus da hepatite B (VHB) constitui um importante problema de Saúde Pública na Amazônia brasileira, onde a exposição precoce, durante a infância, ocorre em áreas de maior endemicidade. Com a finalidade de contribuir para as políticas regionais de controle do VHB na região, foi conduzido um inquérito de prevalência de marcadores sorológicos e moleculares do VHB entre 1.460 gestantes atendidas pelo Programa Pré-Natal, nas nove sub-regiões do Estado do Amazonas, Brasil. Entre essas subregiões, a prevalência do antígeno de superfície (HBsAg) variou de 0 por cento a 8,7 por cento; dos anticorpos anti-core (anti-HBc), de 5,3 a 75,9 por cento;e de anticorpos anti-superfície (anti-HBs), de 10,6 a 73,4 por cento. Entre as 46 gestantes reativas para o HBsAg, 36 (78,3 por cento) foram positivas para VHB-DNA na reação em cadeia da polimerase (PCR). A carga viral de VHB-DNA foi menor que 1x103 cópias/ml em 73,9 por cento das gestantes HBsAg-reativas; porém, 8,7 por cento apresentavam níveis superiores a 1x105 cópias/ml, indicando infecção ativa. Os resultados encontrados mostram sub-regiões do Amazonas com elevada prevalência de VHB entre mulheres grávidas e, embora a maioria apresente baixa viremia, algumas podem representar risco potencial de transmissão mãe-filho, devido à elevada carga viral

Métodos de triagem para malária, em doadores de sangue de área endêmica / Screening methods for malaria, in blood donors in endemic area

Relizou-se um estudo controlado para comparar a presença parasitemiaa e anticorpos antiplasmódios em doadores expostos ao risco de infecçäo malárica, segundo os critérios fixados nas Normas Técnicas em Hemoterapia, do Miníterio da Saúde. Eles estabelecem a rejeiçäo dos candidatos à doaçäo que apresentaram quadro febril há 30 dias e malária há 12 meses, ou que frequentaram área de malária há 6 meses. Foram estudados 395 candidatos incluídos nos critérios de rejeiçäo (expostos) e 383 candidatos aptos (controles), selecionados na triagem de doadores do Hemocentro de Manaus (AM). Em ambos os grupos, realizou-se a gota espessa e o teste da laranja acridina (QBC) para o diagnóstico parasitológico e a imunofluorescência indireta para a pesquisa de anticorpos IgM e IgG ao Plamodium vivax e falciparum. Observou-se diferença significante entre expostos e controles em relaçäo à presença de parasitemia, porém mäo para a presença de anticorpos antiplasmódios (P<0.05). O QBC foi mais sensível do que a gota espessa para detectar parasitemia e a concordância entre ambos foi de 0.66 ao Indice Kappa