Emergence of New Epidemiological Hepatitis B and C Profiles in High Risk Groups in Latin America

Latin America includes Mexico, the islands of the Caribbean and Central and South America, which possess a rich cultural and natural heritage. A narrative literature review was made to determine epidemiological hepatitis B and C profiles in high risk groups in Latin America, such as, drug users, hemophiliacs, and chronic kidney disease (CKD), human immunodeficiency virus (HIV) infected individuals. Using data from international databases that disseminate published quality studies. All studies with desired information regarding site and study population were included. It was observed that HBV prevalence diminished in several groups, probably due to implementation of HBV vaccination in various Latin America Countries (LACs). On the other hand, HCV prevalence is high among high risk groups compared to general population, but different values were observed in LAC, probably due to different access to education programs, assays evaluated, population size and type of recruitment. Due to chronicity of HBV and HCV, it is important to increase access to diagnosis, HBV vaccination and implementation of education programs to high risk groups to diminish burden of these infections

Accuracy of the Zika IgM Antibody Capture Enzyme-Linked Immunosorbent Assay from the Centers for Disease Control and Prevention (CDC Zika MAC-ELISA) for Diagnosis of Zika Virus Infection

Serological diagnosis of Zika virus (ZIKV) infection is challenging because of antigenic cross-reactivity with dengue virus (DENV). This study evaluated the accuracy of the Zika IgM antibody capture enzyme-linked immunosorbent assay (CDC Zika IgM MAC-ELISA) in differentiating between ZIKV and DENV infections. To determine sensitivity, we used acute- and convalescent-phase sera from 21 patients with RT-PCR-confirmed ZIKV infection. To determine specificity, we used acute- and convalescent-phase sera from 60 RT-PCR-confirmed dengue cases and sera from 23 blood donors. During the acute-phase of the illness, the assay presented a sensitivity of 12.5% (2/16) for samples collected 0–4 days post symptoms onset (DPSO), and of 75.0% (3/4) for samples collected 5–9 DPSO. During the convalescent-phase of the illness, the test sensitivity was 90.9% (10/11), 100% (2/2), and 0% (0/2) for samples obtained 12–102, 258–260, and 722–727 DPSO, respectively. Specificity for acute- and convalescent-phase samples from RT-PCR-confirmed dengue cases was 100% and 93.2%, respectively. Specificity for blood donor samples was 100%. The assay is an accurate method for Zika serological diagnosis and proved to be reliable for use during surveillance and outbreak investigations in settings where ZIKV and DENV cocirculate

Desenvolvimento de testes de detecção e quantificação do vírus da Hepatite B em amostras de soro e fluido oral

Made available in DSpace on 2015-10-21T12:25:06Z (GMT). No. of bitstreams: 2 moyra_portilho_ioc_mest_2013.pdf: 2657721 bytes, checksum: ac38c443da941a1c7b8e800b3c858856 (MD5) license.txt: 1748 bytes, checksum: 8a4605be74aa9ea9d79846c1fba20a33 (MD5) Previous issue date: 2015-04-14Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Rio de Janeiro, RJ, Brasil.A detecção e quantificação do DNA do vírus da hepatite B (HBV) são importantes para o diagnóstico da infecção, definição e monitoramento do tratamento antiviral. Entretanto o diagnóstico molecular é difícil em áreas remotas ou com poucos recursos devido ao custo dos métodos comerciais e pouca infra-estrutura para coleta, armazenamento e transporte de amostras de sangue. O objetivo deste estudo foi desenvolver um método para quantificação do DNA do HBV em amostras de soro e fluido oral, e avaliar um método qualitativo \201Cin house\201D para detecção do DNA do HBV em comparação com métodos comerciais disponíveis no mercado. Amostras pareadas de soro e fluido oral foram obtidas de 116 indivíduos, onde 66 eram reagentes para HBsAg no soro e 50 não apresentavam nenhum marcador sorológico no soro. As amostras de soro foram submetidas a testes imunoenzimáticos para detecção de marcadores sorológicos do HBV (HBsAg, anti-HBc, anti-HBs, anti-HBcIgM, anti-HBe e HBeAg) e ao PCR em tempo real para quantificação do DNA do HBV (COBAS TaqMan HBV, Roche). O DNA do HBV foi extraído utilizando o conjunto de reagentes \201CHigh Pure Viral Nucleic Acid kit\201D (Roche Diagnostics, EUA) em amostras de soro e \201CRTP® DNA/RNA Virus Mini kit\201D (Invitek, Alemanha) para amostras de fluido oral Para a detecção qualitativa do HBV foi realizada uma PCR com iniciadores para o gene do core, e para detecção quantitativa do HBV foi utilizada a metodologia de PCR em tempo real com sondas TaqMan® na plataforma Line-Gene 9600 (Bioer Serves Life, Canada). Para obtenção da curva de quantificação foi construído um plasmídeo recombinante obtido a partir de amostras padrão do painel de quantificação do HBV (Optiquant HBV, Acrometrix, Life Technologis, EUA). As seguintes condições da PCR em tempo real foram avaliadas: concentração de DNA (5 e 7,5\03BCL), temperatura de hibridização (60°C e 62°C), e números de ciclos (40 e 45 ciclos). A sensibilidade analítica da PCR em tempo real foi estimada em 10 cópias de HBV DNA/mL e a faixa de quantificação abrangeu cerca de 8 logs de 10. Para quantificação do DNA do HBV em soro e fluido oral foi necessário o aumento da temperatura de hibridização, e para o fluido oral também foi necessário o aumento da concentração de DNA (7,5 \03BCL) e do número de ciclos (45). Entre as amostras de soro HBsAg reagentes, 64 foram quantificadas pela PCR em tempo real comercial e 28 pelo teste quantitativo \201Cin house\201D com carga viral média igual a 3,993 ± 1,922 e 3,761± 1,829 log cópias de HBV DNA/mL (r=0,7643; p<0,0001), respectivamente, e concordância de 37,87% Por outro lado, 8 amostras de fluido oral amplificaram pelo método quantitativo \201Cin house\201D, com carga viral média igual a 4,459 ± 1,127 log cópias de HBV DNA/mL (r=- 0,2994; p= 0,9453). A PCR qualitativa \201Cin house\201D foi capaz de detectar o DNA do HBV em 50 amostras de soro HBsAg reagentes apresentando 75% de concordância com o teste comercial quantitativo (p=1,000), porém somente uma amostra de fluido oral foi detectada por este método. Concluímos que as metodologias qualitativa e quantitativa para detecção do DNA do HBV analisadas neste estudo apresentaram boa eficiência em amostras de soro, porém não foram eficientes em amostras de fluido oral. Estas metodologias podem ser bastante úteis para detecção molecular do HBV em áreas com recursos limitadosThe detection and quantification of the DNA of Hepa titis B virus (HBV) are important to diagnose the infection, determine and monitore the antiviral treatment. However, the molecular diagnosis is difficult in remote areas or presenting low resources, because of the cost of commercial methods and few infrastructu re for collection, storage and transport of blood samples. The objective of this s tudy was to develop a method for quantification of HBV DNA in serum and oral fluid s amples, and to evaluate an in house qualitative method for HBV DNA detection compared t o commercial methods available in the market. Paired serum and oral fluid were obtain ed from 116 individuals, where 66 were HBsAg reactive in their sera and 50 did not pr esent any HBV serological marker in sera. Serum samples were submitted to enzyme immuno assays for detection of HBV serological markers (HBsAg, anti-HBc, anti-HBs, ant i-HBcIgM, HBeAg and anti-HBe) and quantified by commercial real time PCR (COBAS® TaqM an HBV Test, Roche Diagnostics, EUA). HBV DNA was extracted using the commercial kit "High Pure Viral Nucleic Acid Kit" (Roche Diagnostics, USA) in serum samples and "RTP ® DNA / RNA Virus Mini Kit" (Invitek, Germany) for oral fluid s amples. For HBV qualitative detection it was employed a PCR using primers for Core gene, and for quantitative detection of HBV it was employed a real time PCR methodology with Ta qMan ® probes in the Line-Gene 9600 (Bioer Serves Life, Canada) platform. To obtai n the quantification curve, a recombinant plasmid was constructed using standard quantification panel of HBV (Optiquant HBV, Acrometrix, Life Technologis, USA). The following PCR conditions were evaluated in real time PCR: DNA concentration (7.5 and 5 μL), annealing temperature (60° C and 62° C), number of cycles (cycles 40 and 45). The analytical sensitivity of real- time PCR was estimated at 10 HBV DNA copies/mL and the quantitation range comprised about 8 logs of 10. For quantification of HBV DNA in serum and oral fluid, it was necessary to increase the annealing temperature , and for oral fluid, it was also necessary to increase the concentration of DNA (7.5 μL) and the number of cycles (45). Among HBsAg reactive serum samples, 64 were quantif ied by commercial real time PCR and 28 by “in house” quantitative test, with mean v iral load equal to 3,993 ± 1,922 e 3,761± 1,829 log copies of HBV DNA/mL (r=0,7643; p< 0,0001), respectively, and concordance of 66.6%. Moreover, eight oral fluid sa mples were amplified by quantitative "in house" method, with average viral load equal to 4,459 ± 1,127 log copies of HBV DNA/mL (r=-0.2994, p=0,9453). The qualitative "in h ouse" PCR was able to detect HBV DNA in 50 HBsAg reactive serum samples, showing 75% of agreement with the commercial test (p=1.000), but only 1 oral fluid sa mple has been detected by this method. We concluded that the qualitative and quant itative methods for the detection of HBV DNA analyzed in this study presented good effic iency in serum samples, but they were not effective among oral fluid samples. These methodologies can be useful for molecular detection of HBV in areas with limited re source

Applicability of Oral Fluid and Dried Blood Spot for Hepatitis B Virus Diagnosis

Submitted by Sandra Infurna ([email protected]) on 2019-08-27T15:51:28Z No. of bitstreams: 1 GeaneFlores_HelenaCruz_etal_IOC_2019.pdf: 1644866 bytes, checksum: a657dc768b5410107e82a737c8a12990 (MD5)Approved for entry into archive by Sandra Infurna ([email protected]) on 2019-08-27T15:58:16Z (GMT) No. of bitstreams: 1 GeaneFlores_HelenaCruz_etal_IOC_2019.pdf: 1644866 bytes, checksum: a657dc768b5410107e82a737c8a12990 (MD5)Made available in DSpace on 2019-08-27T15:58:16Z (GMT). No. of bitstreams: 1 GeaneFlores_HelenaCruz_etal_IOC_2019.pdf: 1644866 bytes, checksum: a657dc768b5410107e82a737c8a12990 (MD5) Previous issue date: 2019Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Hepatites Virais. Rio de Janeiro, RJ, Brasil.Instituto Federal de Educação, Ciência e Tecnologia do Ceará. Campus Fortaleza. Fortaleza, CE, Brasil.Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Hepatites Virais. Rio de Janeiro, RJ, Brasil.Fundação Oswaldo Cruz. Instituto Gonçalo Muniz. Salvador, BA, Brasil.Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Hepatites Virais. Rio de Janeiro, RJ, Brasil.Hepatitis B virus (HBV) is one of the major causes of chronic liver disease worldwide; however most of individuals are not aware about the infection. Oral fluid and dried blood spot (DBS) samples may be an alternative to serum to HBV diagnosis to increase the access to diagnosis in remote areas or high-risk groups. The main objective of this review is to give an insight about the usefulness of oral fluid and DBS for detecting HBV markers. Several groups have evaluated the detection of HBsAg, anti-HBc, and anti-HBs markers in oral fluid and DBS samples demonstrating 13 to 100% of sensitivity and specificity according different groups, sample collectors, and diagnosis assays. In the same way, HBV DNA detection using oral fluid and DBS samples demonstrate different values of sensitivity according type of collection, studied group, extraction, and detection methods. Thus, serological and molecular diagnostic tests demonstrated good performance for detecting HBV using oral fluid and DBS according some characteristics and could be useful to increase the access to the diagnosis of HBV

Update on hepatitis B and C virus diagnosis

Submitted by sandra infurna ([email protected]) on 2016-05-24T11:37:08Z No. of bitstreams: 1 leticia2_scallioni_etal_IOC_2015.pdf: 1136205 bytes, checksum: bba3b1ce61d8f9bc835a25c8f1ceb040 (MD5)Approved for entry into archive by sandra infurna ([email protected]) on 2016-06-02T12:55:21Z (GMT) No. of bitstreams: 1 leticia2_scallioni_etal_IOC_2015.pdf: 1136205 bytes, checksum: bba3b1ce61d8f9bc835a25c8f1ceb040 (MD5)Made available in DSpace on 2016-06-02T12:55:21Z (GMT). No. of bitstreams: 1 leticia2_scallioni_etal_IOC_2015.pdf: 1136205 bytes, checksum: bba3b1ce61d8f9bc835a25c8f1ceb040 (MD5) Previous issue date: 2015Made available in DSpace on 2016-06-03T12:34:15Z (GMT). No. of bitstreams: 2 leticia2_scallioni_etal_IOC_2015.pdf: 1136205 bytes, checksum: bba3b1ce61d8f9bc835a25c8f1ceb040 (MD5) license.txt: 2991 bytes, checksum: 5a560609d32a3863062d77ff32785d58 (MD5) Previous issue date: 2015Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Hepatites Virais. Rio de Janeiro, RJ, Brasil.Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Hepatites Virais. Rio de Janeiro, RJ, Brasil.Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Hepatites Virais. Rio de Janeiro, RJ, Brasil.Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Hepatites Virais. Rio de Janeiro, RJ, Brasil.Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Hepatites Virais. Rio de Janeiro, RJ, Brasil.Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Hepatites Virais. Rio de Janeiro, RJ, Brasil.Viral hepatitis B and C virus (HBV and HCV) are responsible for the most of chronic liver disease worldwide and are transmitted by parenteral route, sexual and vertical transmission. One important measure to reduce the burden of these infections is the diagnosis of acute and chronic cases of HBV and HCV. In order to provide an effective diagnosis and monitoring of antiviral treatment, it is important to choose sensitive, rapid, inexpensive, and robust analytical methods. Primary diagnosis of HBV and HCV infection is made by using serological tests for detecting antigens and antibodies against these viruses. In order to confirm primary diagnosis, to quantify viral load, to determine genotypes and resistance mutants for antiviral treatment, qualitative and quantitative molecular tests are used. In this manuscript, we review the current serological and molecular methods for the diagnosis of hepatitis B and C

Genetic diversity of hepatitis B virus quasispecies in different biological compartments reveals distinct genotypes

Abstract The selection pressure imposed by the host immune system impacts hepatitis B virus (HBV) quasispecies variability. This study evaluates HBV genetic diversity in different biological fluids. Twenty paired serum, oral fluid, and DBS samples from chronic HBV carriers were analyzed using both Sanger and next generation sequencing (NGS). The mean HBV viral load in serum was 5.19 ± 4.3 log IU/mL (median 5.29, IQR 3.01–7.93). Genotype distribution was: HBV/A1 55% (11/20), A2 15% (3/20), D3 10% (2/20), F2 15% (3/20), and F4 5% (1/20). Genotype agreement between serum and oral fluid was 100% (genetic distances 0.0–0.006), while that between serum and DBS was 80% (genetic distances 0.0–0.115). Two individuals presented discordant genotypes in serum and DBS. Minor population analysis revealed a mixed population. All samples displayed mutations in polymerase and/or surface genes. Major population analysis of the polymerase pointed to positions H122 and M129 as the most polymorphic (≥ 75% variability), followed by V163 (55%) and I253 (50%). Neither Sanger nor NGS detected any antiviral primary resistance mutations in the major populations. Minor population analysis, however, demonstrated the rtM204I resistance mutation in all individuals, ranging from 2.8 to 7.5% in serum, 2.5 to 6.3% in oral fluid, and 3.6 to 7.2% in DBS. This study demonstrated that different fluids can be used to assess HBV diversity, nonetheless, genotypic differences according to biological compartments can be observed

Hepadnavirus detected in bile and liver samples from domestic pigs of commercial abattoirs

Made available in DSpace on 2015-05-08T14:08:47Z (GMT). No. of bitstreams: 2 license.txt: 1914 bytes, checksum: 7d48279ffeed55da8dfe2f8e81f3b81f (MD5) Yasmine_vieiraetal_IOC_2014.pdf: 2182858 bytes, checksum: 5dcdda0a27df5a72bf3a2d5044186d63 (MD5) Previous issue date: 2014Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Desenvolvimento Tecnológico em Virologia. Rio de Janeiro, RJ, Brasil.Universidade Federal Rural do Rio de Janeiro. Departamento de Mircobiologia Veterinária e Imunologia. Rio de Janeiro, RJ, Brasil.Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Hepatites Virais. Rio de Janeiro, RJ, Brasil.Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Desenvolvimento Tecnológico em Virologia. Rio de Janeiro, RJ, Brasil.Fundação Oswaldo Cruz. Bio-Manguinhos. Laboratório de Tecnologia de Anticorpos Monoclonais. Rio de Janeiro, RJ, Brasil.Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Hepatites Virais. Rio de Janeiro, RJ, Brasil.Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Desenvolvimento Tecnológico em Virologia. Rio de Janeiro, RJ, Brasil.Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Desenvolvimento Tecnológico em Virologia. Rio de Janeiro, RJ, Brasil.Background: Preliminary studies showed the prevalence of a virus similar to human hepatitis B virus (HBV-like) in swine from farms in China and the molecular evidence of Hepadnavirus infection in domestic pigs herds in Brazil. In this study, we genetically characterize the swine Hepadnavirus strains in swine from slaughterhouses located in certified abattoirs from Rio de Janeiro State, Brazil and evaluate its hepatotropic potential. Results: Bile and liver samples from swine were positive for partial genome amplification (ORF S and ORF C), direct sequencing and viral load quantification. Sequencing of the gene encoding the surface antigen allowed classification of Hepadnavirus into genotypes, similar to HBV genotype classification. Indirect immunofluorescence confirmed the presence of HBsAg antigen in liver tissue sections. Conclusions: So far our data suggest that commercial swine house an HBV-like virus and this relevant finding should be considered in studies on the origin and viral evolution

Evaluation of two commercially available chikungunya virus IgM enzyme-linked immunoassays (ELISA) in a setting of concomitant transmission of chikungunya, dengue and Zika viruses

Submitted by Ana Maria Fiscina Sampaio ([email protected]) on 2019-12-10T17:38:28Z No. of bitstreams: 1 Kikuti M Evaluation...pdf: 366431 bytes, checksum: 068dafd1d3db10c2333615678a271257 (MD5)Approved for entry into archive by Ana Maria Fiscina Sampaio ([email protected]) on 2019-12-10T17:51:29Z (GMT) No. of bitstreams: 1 Kikuti M Evaluation...pdf: 366431 bytes, checksum: 068dafd1d3db10c2333615678a271257 (MD5)Made available in DSpace on 2019-12-10T17:51:29Z (GMT). No. of bitstreams: 1 Kikuti M Evaluation...pdf: 366431 bytes, checksum: 068dafd1d3db10c2333615678a271257 (MD5) Previous issue date: 2019-01-05Brazilian National Council for Scientific and Technological Development (grants 400830/2013-2 and 440891/2016-7 to GSR; and scholarships to UK, MGR, and GSR); the Bahia Foundation for Research Support (grants PET0026/ 2013, APP0044/2016, and PET0022/2016 to GSR); the Coordination for the Improvement of Higher Education Personnel, Brazilian Ministry of Education (grant 88887.130746/2016-00 to GSR and scholarship to MK); the Oswaldo Cruz Foundation; the Federal University of Bahia; and the Department of Science and Technology, Secretariat of Science, Technology and Strategic Inputs, Brazilian Ministry of Health.Fundação Oswaldo Cruz. Instituto Gonçalo Moniz. Salvador, BA, Brasil / Universidade Federal da Bahia. Salvador, BA, Brasil.Fundação Oswaldo Cruz. Instituto Gonçalo Moniz. Salvador, BA, Brasil / Instituto de Biología Subtropical. Nodo Iguazú. Puerto Iguazu, Misiones, Argentina.Fundação Oswaldo Cruz. Instituto Gonçalo Moniz. Salvador, BA, Brasil.Fundação Oswaldo Cruz. Instituto Gonçalo Moniz. Salvador, BA, Brasil.Fundação Oswaldo Cruz. Instituto Gonçalo Moniz. Salvador, BA, Brasil.Universidade Federal da Bahia. Salvador, BA, Brasil.University of Texas Medical Branch. Galveston, TX, USA.Fundação Oswaldo Cruz. Instituto Gonçalo Moniz. Salvador, BA, Brasil / Universidade Federal da Bahia. Salvador, BA, Brasil.Fundação Oswaldo Cruz. Instituto Gonçalo Moniz. Salvador, BA, Brasil / Emory University. Atlanta, GA, USA.Fundação Oswaldo Cruz. Instituto Gonçalo Moniz. Salvador, BA, Brasil / Universidade Federal da Bahia. Salvador, BA, Brasil.To evaluate the diagnostic performance of the Inbios (Seattle, US) and Euroimmun (Luebeck, Germany) chikungunya virus (CHIKV) IgM enzyme-linked immunoassays (ELISAs). Methods: We evaluated the tests’ accuracy on sera from 372 patients enrolled in an acute febrile illness surveillance study performed in Salvador, Brazil from Sept/2014 to Jul/2016, a period of simultaneous CHIKV, dengue (DENV), and Zika (ZIKV) virus transmission. We assessed the sensitivity on acute and paired convalescent sera from RT-PCR-confirmed CHIKV cases (collected at median one and 19 days postonset of symptoms, respectively), and the specificity on sera of RT-PCR-confirmed DENV and ZIKV cases, and on negative patients. Results: The Inbios and Euroimmun tests’ sensitivities for acute samples were 4.0% and 10.3%, while for convalescent samples they were 92.4% and 96.9%, respectively. Overall, Inbios IgM ELISA specificities for acute and convalescent samples were 97.7% and 90.5%, respectively, and Euroimmun specificities were 88.5% and 83.9%, respectively. Conclusions: Both tests presented high sensitivity for convalescent samples. However, the Euroimmun test returned more equivocal results and presented a slightly lower specificity, which might result in a higher rate of false positives if the test is used in scenarios of low CHIKV transmission, when the chance of CHIKV infection is lower