Feasibility of dried blood spot for hepatitis C diagnosis in vulnerable subjects and people living in remote areas from Brazil.

BACKGROUND: Agile, accessible and cheap diagnosis of hepatitis C virus (HCV) infection is essential to achieve the elimination of this infection, worldwide, as mandated by the World Health Organzation as part of its strategy for 2030. Dried blood spots (DBS) can be an attractive alternative for sample collection among people living in remote areas and vulnerable populations due to the less invasive collection, its biosafety, and storage & transportation of samples at room temperature. DESIGN: This study aims to estimate the usefulness of dried blood spot samples for the diagnosis and the assessment of HCV infection rates in three different settings in Brazil. Cross-sectional analysis of a sample collection from different populations, aiming to assess the performance of the testing algorithms and respective procedures among different populations with diverse background infection rates. METHODS: We reported the evaluation of DBS as alternative samples for detecting anti-HCV in different groups in real life conditions: (I) Vulnerable subjects living in remote areas of Southeast, North and Northeast Brazil (n = 1464); (II) Beauticians (n = 288); (III) People who use non-injectable drugs (n = 201); (IV) patients referred to outpatient care (n = 275). RESULTS: General assay accuracy was 99%, with a weighted kappa value of 0.9, showing an excellent performance. Sensitivities ranged from 87.5% to 100.0% between groups and specificities were above 99.2%. A total of 194 individuals had HCV RNA in serum and concordance of anti-HCV detection in DBS was 98.4%. CONCLUSIONS: DBS samples could be used for anti-HCV detection in different populations recruited in real life conditions and ambulatory settings, with a high overall sensitivity and specificity.

Condições socioeconômicas e impactos da pandemia da Covid-19 na região da Sub-Bacia do Canal do Cunha, Rio de Janeiro / Socioeconomic conditions and impacts of the COVID-19 pandemic in the Canal do Cunha Sub-Basin region, Rio de Janeiro municipality

RESUMO O Município do Rio de Janeiro (MRJ) estava entre as cidades com altas taxas de mortalidade ao longo da pandemia de Covid-19. Neste estudo, analisamos as taxas de incidência, de mortalidade e letalidade por Covid-19 nas áreas com predominância de Aglomerados Subnormais (ASN). Foram considerados todos os 36 bairros da Sub-Bacia do Canal do Cunha (SBCC) associadas às características demográficas, socioeconômicas e epidemiológicas, com estatística espacial de Moran. A taxa de incidência nos bairros da SBCC foi de 621,5/10.000 habitantes. Complexo do Alemão, Mangueira, e Maré tiveram maiores proporções de casos e mortes. A menor incidência (33,6/10.000 habitantes) e mortalidade (8,3/10.000 habitantes), mas com maior taxa de letalidade (24,7%) foi registrada no Complexo do Alemão. Foi observado correlação negativa entre a taxa de mortalidade e a proporção de habitantes nos bairros com ASN (rho= -0,433; p=0,023). Na estatística espacial, houve correlação inversa para a incidência da Covid-19 (índice Moran, -0,155863; p=0,02). Conclui-se que incidência e mortalidade nas áreas de ASN estão significativamente relacionadas com as estruturas sociodemográficas, demandando o reforço dos sistemas de vigilância e de controle da Covid-19 em territórios de favelas. As recomendações não farmacológicas e a Atenção Primária à Saúde em favelas desempenham relevante papel na redução da transmissão, mortalidade e iniquidades em saúde.

ABSTRACT Along COVID-19 pandemics, Rio de Janeiro (MRJ) has figured among the Brazilian cities with the highest mortality rates. In this study, we explored COVID-19 incidence and mortality in areas with a predominance of subnormal population clusters (SNC) in MRJ. We analyzed 36 neighborhoods of Canal do Cunha Sub-basin (CCSB) and the associations between demographic, socioeconomic, and epidemiological features, and the cumulative incidence, mortality, and lethality rates, and Moran's spatial statistics were performed. The incidence rate in CCSB neighborhoods was 621.5/10,000 inhabitants. Complexo do Alemão, Mangueira, and Maré had the highest proportions of cases and deaths. And the lowest incidence (33.6/10,000 inhabitants), mortality (8.3/10,000 inhabitants), but with the highest lethality rate (24.7%) was recorded in Complexo do Alemão. There was a negative correlation between mortality and the proportion of inhabitants in neighborhoods with ASN (rho= -0.433; p=0.023). In spatial statistics, there was an inverse correlation for the incidence (Moran index, -0.155863; p=0.02). It is concluded that incidence and mortality in SNC areas are significantly related to their sociodemographic structures, highlighting the need to strengthen the surveillance and control systems of COVID-19 in slums territories. Non-pharmacological recommendations and suitable Primary Health Care in slums areas play a relevant role in reducing viral transmission, mortality, and health inequities.

Performance of HCV Antigen Testing for the Diagnosis and Monitoring of Antiviral Treatment: A Systematic Review and Meta-Analysis.

BACKGROUND AND AIMS: Active hepatitis C virus (HCV) infection is based on the detection of HCV RNA that it is effective but presents high cost and the need to hire trained personnel. This systematic review and meta-analysis is aimed at evaluating the diagnostic accuracy of HCV Ag testing to identify HCV cases and to monitor antiviral treatment including DAA treatment. METHODS: The studies were identified through a search in PubMed, Lilacs, and Scopus from 1990 through March 31, 2020. Cohort, cross-sectional, and randomized controlled trials were included. Two independent reviewers extracted data and assessed quality using an adapted Quality Assessment of Diagnostic Accuracy Studies (QUADAS-2) tool. Our primary outcome was to determine the accuracy of HCV Ag detection for the diagnosis, which we estimated using random-effects meta-analysis. RESULTS: Of 3,062 articles identified, 54 met our eligibility criteria. The studies described cohorts from 20 countries, including 14,286 individuals with chronic HCV individuals. Studies for ECLIA technology demonstrated highest quality compared to studies that used ELISA. The pooled sensitivity and specificity (95% CI) for HCV Ag detection of active HCV infection were 98.82% (95%CI = 98.04%; 99.30%) and 98.95% (95%CI = 97.84%; 99.49%), respectively. High concordance was found between HCV Ag testing and HCV RNA detection 89.7% and 95% to evaluate antiviral treatment. CONCLUSIONS: According to our findings, HCV Ag testing could be useful to identify HCV active cases in low-resource areas. For antiviral treatment, HCV Ag testing will be useful at the end of treatment.

Dried blood spot sampling as an alternative for the improvement of hepatitis B and C diagnosis in key populations.

BACKGROUND: To achieve the elimination of hepatitis B and C, there is an urgent need to develop alternative strategies to increase the access of diagnosis, particularly among key populations such as people living with human immunodeficiency virus (HIV), individuals with coagulopathies and chronic kidney disease (CKD) patients. AIM: To evaluate the use of dried blood spot (DBS) in the detection of hepatitis B virus (HBV) and hepatitis C virus (HCV) markers. METHODS: A total of 430 individuals comprised of people living with HIV, coagulopathies and CKD provided paired serum and DBS samples. HBsAg, anti-HBc and anti-HCV were tested in those samples using a commercial electrochemiluminescence. Demographic and selected behavioral variables were evaluated to assess possible association with HBV and HCV positivity. RESULTS: Using DBS, HBsAg prevalence varied from 3.9% to 22.1%, anti-HBc rates varied from 25.5% to 45.6% and anti-HCV positivity ranged from 15.9% to 41.2% in key populations. Specificities of HBV and HCV tests using DBS varied from 88.9% to 100%. The HBsAg assay demonstrated the best performance in CKD and coagulopathy individuals and the anti-HCV test had a sensitivity and specificity of 100% in people living with HIV. Accuracy of HBV and HCV detection in DBS varied from 90.2% to 100%. In the CKD group, HBsAg positivity was associated with infrequent use of condoms, and anti-HBc positivity was associated with sharing nail cutters/razors/toothbrushes. Anti-HCV reactivity was positively associated with a history of transplantation and length of time using hemodialysis in both specimens. In people living with HIV, only the male gender was associated with anti-HBc positivity in serum and DBS. CONCLUSION: DBS with electrochemiluminescence are useful tools for the diagnosis and prevalence studies of hepatitis B and C among key populations and may increase the opportunity to foster prevention and treatment.

Usefulness of automated assays for detecting hepatitis B and C markers in dried blood spot samples.

OBJECTIVE: Dried blood spots (DBSs) can be used as an alternative to serum samples because they are easily collected and can be transported without refrigeration to reference laboratories for diagnosis. The present study was performed to evaluate the utility of electrochemiluminescence immunoassay "ECLIA" for anti-HCV, HBsAg and anti-HBc detection from DBS samples. RESULTS: Anti-HCV was detected in 103 DBS samples from 108 paired, positive serum and undetected in 364 DBS samples from 366 paired, negative specimens, giving a sensitivity of 95.4% and a specificity of 99.4%. HBsAg was detected in 67 DBS samples out of 71 positive, paired serum and was undetected among 295 DBS samples from 298 paired, negative specimens, giving a sensitivity and specificity of 94.4% and 99%, respectively. Anti-HBc was detected in 160 DBS samples from 185 paired, positive serum specimens and undetected in 349 DBS samples from 357 paired, negative serum specimens, giving a sensitivity of 86.5% and a specificity of 97.8%. Overall, the Kappa index indicated a high agreement between results obtained for the serum and DBS samples (k: 0.95, 0.93 and 0.86 for anti-HCV, HBsAg, anti-HBc, respectively). In conclusion, the ECLIA test could be used for detecting hepatitis B and C markers in DBS.

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

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.

Assessing hepatitis B immunity using dried blood spot samples from HIV+ individuals.

This study aims to evaluate the utility of an optimized enzyme immunoassay (EIA) to detect and quantify antibodies against hepatitis B surface antibody (anti-HBs) in dried blood spots (DBSs) within the context of human immunodeficiency virus (HIV) status. Serum and DBS samples were obtained from 56 HIV+ and 99 HIV- patients and subjected to EIA for the detection of anti-HBs, where sample volume and cut off value were modified for DBS testing. Sensitivities of anti-HBs detection in DBS were 79.8% and 76.8% in HIV- and HIV+ subjects, respectively. Concordant results for anti-HBs in serum and DBS presented high mean CD8 cell counts, HIV viral load and optical density (OD) values of anti-HBs. Anti-HBs titers were significantly higher in serum, whether or not anti-HBs titers were detected in DBS. It was possible to detect anti-HBs in DBS as low as 17.4 and 27.3 IU/mL among HIV+ and HIV- subjects, respectively. In conclusion, DBS can be used to detect and quantify anti-HBs in HIV-infected individuals, which could increase access to diagnosis and vaccination.

Detection of occult hepatitis B in serum and oral fluid samples.

In occult hepatitis B infection (OBI), hepatitis B virus DNA (HBV DNA) can be detected in serum samples; however, oral fluid collection for detection of HBV DNA has not yet been explored, despite the availability of collection devices. Serum and oral fluid samples from 45 hepatitis B core antibody (anti-HBc)-positive patients were collected for the amplification of the HBV polymerase gene. HBV DNA was detected in five serum and four oral fluid samples (the detection limit for oral fluid was 1.656 log IU/mL in paired serum). In conclusion, simple methodologies of sample collection and in-house polymerase chain reaction (PCR) allowed detection of HBV DNA, and these could be used to improve the diagnosis of OBI, especially in locations with limited resources.

Performance of rapid diagnostic tests for detection of Hepatitis B and C markers in HIV infected patients.

There is little information describing the influence of HIV infection upon the performance of rapid diagnostic tests (RDTs) for hepatitis B and C virus diagnosis. This study aims to evaluate the performance of RDTs for HBsAg and anti-HCV detection among HIV-infected individuals. A total of 362 HIV infected individuals were recruited from clinics between January 2013 to November 2014 in the southeast and northeast of Brazil. HBsAg and anti-HCV were detected using commercial EIAs and four RDTs: HBV (Vikia HBsAg® and Wama Imuno-Rapido HBV®) and HCV (Bioeasy Teste Rápido HCV® and Wama Imuno-Rapido HCV®). Reactive HBsAg and anti-HCV serum samples were tested for HBV DNA and HCV RNA. Sensitivity, specificity and kappa statistic were determined. Using EIA, HBsAg and anti-HCV were detected in 14 (3.9%) and 37 (10.2%) serum samples respectively. Using serum only, HBsAg RDTs demonstrated sensitivities and specificities above 92.0% and Kappa values above 89.0%. Anti-HCV RDTs demonstrated sensitivity and specificities above 82.0% and Kappa higher than 89.0%. Using whole blood samples, Vikia HBsAg® and Wama Imuno-Rapido HCV® showed sensitivity and specificity above 99.0% with Kappa of 66.4% and 100%, respectively. HIV viral load was higher among discordant results for anti-HCV RDT. RDTs demonstrated good performance in HIV infected individuals showing the usefulness of assays in this population.

Evaluation of HBsAg and anti-HBc assays in saliva and dried blood spot samples according HIV status.

Influence of HIV status in HBV markers detection in saliva and dried blood spots (DBS) was not well established. This study aims to evaluate the performance of optimized commercial immunoassay for identifying HBsAg and anti-HBc in saliva and DBS according HIV status. A sum of 535 individuals grouped as HIV+, HBV+, HIV/HBV+ and HIV/HBV- were recruited where 347 and 188 were included for HBsAg and anti-HBc evaluation, respectively. Serum, DBS collected in Whatman 903 paper and saliva obtained using salivette device were analyzed using EIA. Increased sample volume and ROC curve analysis for cut off determination were used for DBS and saliva testing. HBsAg detection in saliva and DBS exhibited sensitivities of 80.9% and 85.6% and specificities of 86.8% and 96.3%. Sensitivity of anti-HBc in saliva and DBS were 82.4% and 76.9% and specificities in saliva and DBS were 96.9% and 91.7%. Low sensitivities were observed for HBsAg (62%) and anti-HBc (47%) detection in saliva of HIV/HBV+ individuals. OD values were also lower for HBsAg detection in DBS and saliva of HIV/HBV+ individuals compared to their serum samples. Statistical significance was found for sensitivities in HBsAg detection between saliva and DBS demonstrating high sensitivity for DBS specimens. In conclusion, HIV status or antiretroviral treatment appears to interfere in the performance of HBsAg and anti-HBc detection in DBS and saliva samples using the adapted commercial EIA.

Performance of ANTI-HCV testing in dried blood spots and saliva according to HIV status.

The use of saliva and dried blood spots (DBS) could increase access to HCV diagnosis for high-risk populations, such as HIV-infected individuals, but the performance of these assays has not been well established in this group. This study aims to evaluate HIV status, particularly TCD4+ cell count and viral load, in the performance of anti-HCV testing using DBS and saliva. A total of 961 individuals classified as HCV+, HIV+, or HIV/HCV+, as well as negative controls, donated serum, DBS, and saliva samples for anti-HCV testing using a commercial enzyme immunoassay. Sample volume was modified for DBS and saliva, and an ROC curve was used for cut-off determination in saliva. Anti-HCV sensitivities were greater than 93% using DBS and saliva in the HCV+ group, while they were 83.3% and 95.6% for HCV/HIV+ individuals for DBS and saliva assays, respectively. Specificity varied from 91.7% to 100% using saliva and DBS in HIV monoinfected and control subjects. When only anti-HCV/HCV RNA+ serum samples, that is, true positives, were considered, the sensitivities were 98.3% and 100% for DBS and saliva, respectively, in the HCV+ group and 91.6% and 94.8% for DBS and saliva, respectively, in the HIV/HCV+ group. High absorbance values were observed among those presenting with HCV RNA in serum and low HIV viral load (less than 50 copies/mL). In conclusion, DBS and saliva samples could be used for anti-HCV detection, particularly to identify active HCV cases, but low sensitivity was observed for anti-HCV testing using DBS in the HIV/HCV+ group.

Avaliação da reconstituição imune em gestantes submetidas à quimioprofilaxia para prevenção da transmissão vertical do HIV-1 e do impacto imunológico da descontinuidade do tratamento no pós-parto / Assessment of immune reconstitution in women undergoing chemoprophylaxis to prevent vertical transmission of HIV-1 and immunological impact of discontinuation of treatment postpartum

Nos últimos anos, a epidemia de aids no Brasil tem mostrado mudanças significativas em seu perfil, caracterizado principalmente pela sua feminização. O crescimento de casos de aids em mulheres, especialmente aquelas em idade reprodutiva, tem resultado em aumento da transmissão vertical do HIV-1. Portanto, a prevenção da transmissão vertical do HIV-1 é um grande desafio a ser enfrentado. Vários estudos têm mostrado uma redução da transmissão vertical do HIV-1 ((menor que)2(por cento)), com uso adequado de terapia anti-retroviral (TARV) durante a gravidez e o parto. Além disso, o uso da TARV está diretamente associado a uma significativa recuperação do sistema imunológico, com um aumento quantitativo dos níveis periféricos de células T CD4 (mais) , acompanhado pela reversão de processos de infecção, tais como ativação celular persistente, e conseqüentemente, mudanças nos perfis de diferenciação celular. Este estudo visa investigar o perfil imunológico, em particular a reconstituição imunológica em mulheres grávidas infectadas pelo HIV-1, em diferentes momentos da TARV ou profilaxia da transmissão vertical, após o parto e descontinuidade do tratamento. Trinta e sete mulheres infectadas pelo HIV-1 e virgens de tratamento, recrutados no Hospital Geral de Nova Iguaçu-MS, foram avaliadas antes do início da TARV (Pré-TARV) parto (TARV) e seis meses ou um ano após o parto (Pós-TARV). Nestes três momentos, as amostras de sangue das voluntárias foram coletadas para obter células mononucleares do sangue periférico, os quais foram criopreservados e posteriormente analisadas para diferentes fenótipos por citometria de fluxo. Com a TARV, o controle da replicação viral foi obtido, com um aumento significativo de células T CD4 (mais) . Variações percentuais nos níveis circulantes de células T naïve, memória central e memória efetoras em ambas subpopulações, TCD4 (mais) e CD8 (mais) , não foram detectados ao longo do estudo. Uma ativação celular persistente, traduzida por níveis elevados de linfócitos T CD8 (mais) /CD38 (mais) , foi observada nos três momentos analisados, inclusive no momento do parto, quando o controle da carga viral ocorre. Portanto, a ativação celular em mulheres grávidas sob TARV não está associado com a replicação viral, mas a outros fatores inerentes à gravidez e ao parto. Importante para a prevenção da transmissão vertical do HIV-1, a TARV não induziu alterações significativas na circulação das subpopulações celulares nesses pacientes, e sua suspensão não causou impacto nos perfis das células estudadas, pelo menos no curto prazo.