RESUMEN
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.
Asunto(s)
Hepacivirus , Hepatitis C , Humanos , Hepacivirus/genética , Brasil/epidemiología , Estudios Transversales , Estudios de Factibilidad , Poblaciones Vulnerables , ARN Viral , Pruebas con Sangre Seca/métodos , Hepatitis C/diagnóstico , Hepatitis C/epidemiología , Sensibilidad y EspecificidadRESUMEN
Little information is available regarding the prevalence of viral hepatitis in Central West Argentina. This study aims to give new information regarding HBV and HCV prevalence, genotypes, and risk factors in Central West Argentina and the suitability of dried blood spot (DBS) sampling for HBV and HCV screening. METHODS: A total of 622 individuals were included; the mean age was 36.6 ± 14.3 years and 55.4% were females. HBV and HCV markers were detected using serological and molecular analysis, and risk factors were evaluated using statistical analysis. RESULTS: Using serum samples, the HBsAg prevalence was 1.8%, the rate of HBV exposure (anti-HBc positivity) was 5.3%, and the rate of HBV immunity was 34.9%. HBV DNA was found in four out of 11 HBsAg+ samples, and the viruses in three of these samples were classified as genotypes A1, A2 and F2a. Multivariate analysis showed that anti-HBs positivity was associated with the level of schooling and history of HBV vaccination. The anti-HCV prevalence was 2.6%, and HCV RNA was found in 11 samples, seven of which contained viruses of genotypes 1a (n = 2), 1b (n = 3) and 2 (n = 2). The sensitivity of the DBS assay for HBsAg, anti-HBc, and anti-HCV was 100%, 66.6%, and 75%, respectively, and the specificity was above 98% for all markers when compared to serum. CONCLUSION: A low rate of HBV immunity was observed, demonstrating the importance of HBV vaccination. High HCV prevalence was found, and HCV 1b was closely related to other Argentinian isolates. Finally, the performance of DBS testing in this population needs more optimization to increase its sensitivity and specificity.
Asunto(s)
Hepacivirus/aislamiento & purificación , Virus de la Hepatitis B/aislamiento & purificación , Hepatitis B/epidemiología , Hepatitis B/virología , Hepatitis C/epidemiología , Hepatitis C/virología , Adulto , Argentina/epidemiología , Estudios Transversales , Femenino , Genotipo , Hepacivirus/clasificación , Hepacivirus/genética , Hepatitis B/sangre , Hepatitis B/diagnóstico , Anticuerpos contra la Hepatitis B/sangre , Virus de la Hepatitis B/clasificación , Virus de la Hepatitis B/genética , Hepatitis C/sangre , Hepatitis C/diagnóstico , Humanos , Masculino , Persona de Mediana Edad , Filogenia , Prevalencia , Adulto JovenRESUMEN
BACKGROUND: Hepatitis B virus (HBV) testing in oral fluid samples may provide advantages in diagnosis, screening or prevalence studies, especially among individuals with venous access difficulties. This study aims to optimize one commercially available assay for detecting total anti-HBc marker in oral fluid samples and to evaluate its utility under real life conditions in different settings for the purposes of prevalence and diagnostic studies. METHODS: Oral fluid was collected using a Salivette device and some parameters were initially evaluated: type of elution buffer and sample volume. Thereafter, the utility of oral fluid samples for detection of anti-HBc was evaluated in real life conditions in which, 1296 individuals gave serum and oral fluid samples. All serum samples were submitted to commercial EIAs to detect total anti-HBc, according to the manufacturer's instructions and oral fluid samples according to previous optimization. RESULTS: In optimization evaluation, PBS/BSA 0.5% and 100 µL of oral fluid (volume was two-fold increased compared to serum in EIA) were chosen as transport buffer and sample volume. In the field study, anti-HBc was detected in 211 out of 1296 serum samples giving overall oral fluid sensitivity of 52.6% and specificity of 96%. Concordance was higher in ambulatory setting (67.7) compared to general population (31.8). Mean ± standard deviation values of optical density/cutoff (OD/CO) in serum samples were higher in false-negative oral fluid samples than those seen in true positive samples. Sensitivity was higher in those presenting active infection compared to anti-HBc isolate and past infection. Sensitivity also increased in the ambulatory group when HCV individuals were excluded. CONCLUSIONS: It was possible to optimize a commercial EIA for detecting anti-HBc in oral fluid samples and where the highest concordance was found in ambulatory settings and among individuals with active infection.
Asunto(s)
Anticuerpos contra la Hepatitis B/análisis , Hepatitis B/diagnóstico , Técnicas para Inmunoenzimas/métodos , Saliva/virología , Adulto , Femenino , Humanos , Masculino , Persona de Mediana Edad , Sensibilidad y EspecificidadRESUMEN
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 immunodeï¬ciency 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.
Asunto(s)
Desecación , Infecciones por VIH , Anticuerpos contra la Hepatitis B/sangre , Virus de la Hepatitis B/inmunología , Hepatitis B/inmunología , Tamizaje Masivo/métodos , Manejo de Especímenes/métodos , Adolescente , Adulto , Anciano , Anciano de 80 o más Años , Femenino , Antígenos de Superficie de la Hepatitis B/inmunología , Humanos , Técnicas para Inmunoenzimas/métodos , Masculino , Persona de Mediana Edad , Sensibilidad y Especificidad , Adulto JovenRESUMEN
BACKGROUND: This study was conducted to determine the prevalence of HBV, HCV, and HDV in urban populations and Amerindians living in the state of Tocantins (Eastern Amazon). METHODS: A total of 948 individuals were recruited in Tocantinopolis city (Tocantins state) of whom 603 were Amerindians (from 6 tribes) and 345 were non-Amerindians (6 urban areas of Tocantinópolis city). Anti-HCV, HBsAg, anti-HBc, anti-HBs, anti-HBc IgM, anti-HBe, HBeAg, and anti-delta antibodies were determined using enzyme immunoassay. RESULTS: HBV cleared infection (both anti-HBc/anti-HBs+), chronic inactive/immune controlled HBV infection (anti-HBc + only), previous HBV vaccination (anti-HBs + only), active HBV infection (HBsAg+), individuals susceptible to HBV, and anti-HCV reactivity were found in 12.9, 1.8, 27.2, 0.5, 57.7, 1.2% in Amerindians and 12.1, 2.0, 37.1, 0.3, 55.4, 0.3% in non-Amerindians respectively. Out of 139 anti-HBc reactive individuals, 70 were anti-HBe reactive and none presented HBeAg or anti-HBc IgM. Anti-HBc prevalence was associated to older age (p < 0.0001). Overall anti-Delta prevalence was 0.3% and regarding anti-HBc reactive individuals, anti-delta prevalence was 3.4 and 0% in Amerindians and non-Amerindians respectively. CONCLUSIONS: Overall low prevalence of HBV and HCV infection was found in the populations studied, but high HBV and HCV prevalence was observed in Amerindians compared to non-Amerindians suggesting that these individuals have a higher likelihood of acquiring to these infections. Anti-delta antibodies were found among Amerindians from Eastern Amazon suggesting a risk for this population. Of note is that nearly half of Amerindians had no anti-HBs, indicating a need for HBV vaccination campaigns in this population.
Asunto(s)
Hepatitis B/epidemiología , Hepatitis C/epidemiología , Hepatitis D/epidemiología , Indígenas Norteamericanos/estadística & datos numéricos , Población Urbana/estadística & datos numéricos , Adolescente , Adulto , Anciano , Anciano de 80 o más Años , Brasil/epidemiología , Niño , Preescolar , Estudios Transversales , Femenino , Hepatitis B/sangre , Anticuerpos contra la Hepatitis B/análisis , Anticuerpos contra la Hepatitis B/sangre , Hepatitis C/sangre , Anticuerpos contra la Hepatitis C/análisis , Anticuerpos contra la Hepatitis C/sangre , Hepatitis D/sangre , Humanos , Lactante , Recién Nacido , Masculino , Persona de Mediana Edad , Prevalencia , Ríos , Adulto JovenRESUMEN
Dried blood spots (DBSs) could be an alternative to serum for hepatitis B virus (HBV) and hepatitis C virus (HCV) diagnosis. This study aims to evaluate two enzyme immunoassays (EIAs) for HBsAg and anti-HCV detection using DBS. Serum was tested using commercial EIA. DBS was tested using optimized EIA developed for serum and commercial EIA developed for DBS (Imunoscreen). Concordances between DBS and serum samples for both markers and EIAs were higher than 97%. Both EIAs demonstrated good performance for HBsAg and anti-HCV detection using DBS, and these methods could be used unchangeably increasing the access for HBV and HCV diagnosis.
Asunto(s)
Pruebas con Sangre Seca , Hepacivirus/aislamiento & purificación , Virus de la Hepatitis B/aislamiento & purificación , Técnicas para Inmunoenzimas/métodos , HumanosRESUMEN
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.
Asunto(s)
Sangre/inmunología , Ensayo de Inmunoadsorción Enzimática/métodos , Infecciones por VIH/complicaciones , Anticuerpos contra la Hepatitis C/análisis , Anticuerpos contra la Hepatitis C/sangre , Hepatitis C/diagnóstico , Saliva/inmunología , Adulto , Anciano , Recuento de Linfocito CD4 , Desecación , Femenino , Humanos , Masculino , Persona de Mediana Edad , Sensibilidad y Especificidad , Manejo de Especímenes/métodos , Encuestas y Cuestionarios , Carga ViralRESUMEN
Rapid tests (RTs) can be used as an alternative method for the conventional diagnosis of hepatitis B virus (HBV). This study aims to evaluate antibodies to HBsAg (anti-HBs) and antibodies to HBeAg (anti-HBe) RTs under different Brazilian settings. The following three groups were included: GI: viral hepatitis outpatient services; GII: low resource areas; and GIII: crack users and beauticians. Imuno-rápido anti-HBsAg™ and Imuno-rápido anti-HBeAg™ RTs were evaluated and showed specificities greater than 95% in all groups. The sensitivity values to anti-HBs were 50.38%, 51.05% and 46.73% and the sensitivity values to anti-HBe were 76.99%, 10.34% and 11.76% in the GI, GII and GIII groups, respectively. The assays had a low sensitivity and high specificity, which indicated their use for screening in regions endemic for HBV.
Asunto(s)
Anticuerpos contra la Hepatitis B/sangre , Antígenos de Superficie de la Hepatitis B/sangre , Antígenos e de la Hepatitis B/sangre , Virus de la Hepatitis B/inmunología , Hepatitis B/diagnóstico , Adulto , Humanos , Persona de Mediana Edad , Juego de Reactivos para Diagnóstico , Sensibilidad y EspecificidadRESUMEN
BACKGROUND: Rapid tests (RTs) might have several advantages over standard laboratory procedures, increasing access to diagnosis, especially among vulnerable populations and/or those living in remote areas. The aim of this study was to evaluate the performance of RTs for the detection of hepatitis B virus surface antigen (HBsAg) in samples from different populations/settings. METHODS: Three RTs for HBsAg detection (Vikia® HBsAg, HBsAg Teste Rápido®, and Imuno-Rápido HBsAg®) and different biological specimens (serum, whole blood, and saliva) were evaluated. Analyses comprised a reference panel and samples from field studies targeting suspected cases of hepatitis B virus (HBV) (G I), individuals living in deprived areas (G II), and highly vulnerable individuals (G III). Enzyme immunoassay (EIA) was defined as the gold standard in this study. Reproducibility, repeatability, and cross-reactivity with other infectious agents such as dengue, immunodeficiency (HIV), and hepatitis C (HCV) viruses and T. pallidum were determined. RESULTS: For the reference panel, the sensitivity and specificity of all HBsAg RTs were higher than 93.00 %. G I presented the highest kappa values for all rapid assays using sera samples. When using serum, the sensitivity values were higher than 93.40 for G I, 60.00 % for G II and 66.77 % for G III, and the specificity values were higher than 99.50 for GI, 97.20 for G II and 99.10 % for G III for all tests. For whole blood samples & the Vikia® HBsAg assay, the best performance was achieved for GIII (k = 79.75 %). For saliva samples, the Imuno-Rápido HBsAg® assay showed the highest concordance values with EIA for G I (40.68 %) and G II (32.20 %). The reproducibility and repeatability of all RTs for serum and saliva were excellent, and the concordance between HBsAg EIAs and RTs using samples reactive with other infectious agents varied from 70.10 % to 100.00 %. CONCLUSIONS: The overall performance of RTs for HBsAg in serum was high/moderately high for all groups, thereby promoting increased access to HBV diagnosis among vulnerable populations as well as samples from individuals in emergency settings or remote areas. Rapid tests for HBsAg using whole blood could be used in prevalence studies, though these assays should not be used for saliva samples.
Asunto(s)
Antígenos de Superficie de la Hepatitis B/sangre , Hepatitis B/diagnóstico , Técnicas para Inmunoenzimas/métodos , Adulto , Reacciones Cruzadas/inmunología , Femenino , Hepacivirus/inmunología , Hepatitis B/sangre , Antígenos de Superficie de la Hepatitis B/análisis , Virus de la Hepatitis B/inmunología , Hepatitis C/diagnóstico , Hepatitis C/virología , Anticuerpos contra la Hepatitis C/sangre , Humanos , Masculino , Persona de Mediana Edad , Reproducibilidad de los Resultados , Sensibilidad y EspecificidadRESUMEN
BACKGROUND: Oral fluid (OF) sample collection and stability for HBsAg detection are not fully established. This study aims to investigate the applicability of OF collectors and sample stability for Hepatitis B virus surface antigen detection. METHODS: Paired serum and OF samples were obtained from 191 individuals, and Chembio (Chembio Diagnostic System, USA) and Salivette (Sarstedt, Germany) devices were used for OF collection. Two HBsAg enzyme immunoassays (EIAs) were used (HBsAg One kit, Radim, Rome, Italy and ETI-MAK-4, DiaSorin, Vercelli, Italy) to determine the most efficient method according OF collector. Sample volume, incubation time, and cutoff (CO) value were evaluated. The stability of OF samples was determined under different environmental conditions. RESULTS: Chembio samples analyzed using DiaSorin EIA without modification of the manufacturer's instructions, demonstrated a sensitivity of 95.24% and a specificity of 100%. Salivette samples analyzed with Radim EIA with receiver operating characteristic (ROC) curve for calculating the CO showed a sensitivity of 78.26% and a specificity of 89.88%. HBsAg was detected in Chembio and Salivette samples under different environmental conditions, but the Chembio samples were the most stable. CONCLUSIONS: Both collectors can be used for HBsAg detection in OF samples, but some modifications of commercial EIAs should be incorporated for Salivette device. OF samples were reliably stable and could be stored for up to 90 days at 2-8°C.
Asunto(s)
Antígenos de Superficie de la Hepatitis B/aislamiento & purificación , Hepatitis B/epidemiología , Saliva , Manejo de Especímenes/métodos , Adolescente , Adulto , Anciano , Brasil , Distribución de Chi-Cuadrado , Niño , Preescolar , Femenino , Hepatitis B/diagnóstico , Hepatitis B/virología , Antígenos de Superficie de la Hepatitis B/sangre , Antígenos de Superficie de la Hepatitis B/química , Humanos , Técnicas para Inmunoenzimas , Masculino , Persona de Mediana Edad , Prevalencia , Curva ROC , Saliva/química , Saliva/virología , Estudios Seroepidemiológicos , Manejo de Especímenes/instrumentación , Temperatura , Factores de TiempoRESUMEN
Immunoglobulin Y (IgY) could be used in serological diagnosis focused on several infectious agents. This study aims to produce IgY anti-hepatitis B virus surface antigen (anti-HBs) and to assess its use in enzyme immunoassays. Antibodies were produced by immunizing chickens with Hepatitis B vaccine associated (group A), or not, with adjuvant CpG-ODN (group B). Eggs were collected for 20 weeks, yolks were purified based on using polyethylene glycol and affinity chromatography. IgY anti-HBs was featured based on SDS-PAGE and Western Blot techniques. Total protein concentration was measured through spectrophotometry. In-house ELISA used to detect HBsAg was developed based on using IgG/HRP conjugate and IgY-anti-HBs sensitized microplates. Thus, IgY anti-HBs were confirmed through molecular pattern based on SDS-PAGE, whereas specificity of anti-HBs was confirmed through Western Blot. Mean total protein reached 3.27 ± 3.00 mg/mL and 3.11 ± 3.12 mg/mL in groups A and B, respectively. In-house ELISA was developed based on using a panel of HBV positive and negative serum samples; it recorded 100 % sensitivity and 78.9 % specificity to detect HBsAg. In conclusion, it was possible producing anti-HBs IgY by immunizing chickens with HBV vaccine; this molecule could be used as capture antibody to help detecting HBsAg in-house ELISA.
Asunto(s)
Antígenos de Superficie de la Hepatitis B , Hepatitis B , Animales , Pollos , Vacunas contra Hepatitis B , Hepatitis B/diagnóstico , Antígenos e de la Hepatitis B , Anticuerpos contra la Hepatitis BRESUMEN
Detection of hepatitis B virus (HBV) serological markers in dried blood spot (DBS) samples by enzyme immunoassay (ELISA) has not yet been fully optimized. In this study, the ability to detect three HBV markers (HBsAg, anti-HBc, and anti-HBs) was evaluated in DBS samples using a modified commercial ELISA. Matched serum and DBS samples were obtained from individuals with or without a past history of HBV infection. Sera samples were tested according to the manufacturer's instructions, but for DBS testing, paper diameters, elution buffer, volume of input sample, and cut-off values were evaluated to optimize the assay. Stability studies were done on DBS stored at for up to 180 days at different temperatures. The absorbance values that yielded the maximum sensitivity and specificity were determined based on the area under the ROC curve (AUROC) and chosen as the cut-off value. Using this parameter, sensitivity was 90.5%, 97.6%, and 78% for anti-HBc, HBsAg, anti-HBs assays, respectively. Specificity was 92.6%, 96.7%, and 97.3% for anti-HBc, HBsAg, and anti-HBs assays, respectively. HBV markers could be detected in DBS samples until 63 days after sample collection at most temperatures, but storage at -20°C yielded more consistent results. These results indicate that modified ELISA can be used to detect HBV markers in DBS samples, particularly if the samples are stored appropriately.
Asunto(s)
Pruebas con Sangre Seca , Anticuerpos contra la Hepatitis B/sangre , Antígenos del Núcleo de la Hepatitis B/inmunología , Antígenos de Superficie de la Hepatitis B/sangre , Virus de la Hepatitis B/aislamiento & purificación , Hepatitis B/diagnóstico , Biomarcadores/sangre , Ensayo de Inmunoadsorción Enzimática , Hepatitis B/inmunología , Antígenos de Superficie de la Hepatitis B/inmunología , Virus de la Hepatitis B/inmunología , Humanos , Sensibilidad y EspecificidadRESUMEN
In this study, a modified enzyme immunoassay (EIA) was evaluated for the Hepatitis B surface antigen (HBsAg) among whole saliva and oral fluid samples. Specimens were collected from 115 individuals who gave serum and oral fluid using Salivette (Sarstedt, Nümbrecht, Germany) and whole saliva. Saliva specimens were tested following a modified ELISA, and the results were compared with paired serum specimens that were tested according to the supplier's instructions. Transport buffer for the oral fluids, sample volume for assay, incubation period of sample with conjugate as well as cut-off values were evaluated to optimize the assay. The highest sensitivity and specificity were obtained by increasing the incubation of sample and conjugate to 16 hr and using the area under the receiver operating characteristic curve to calculate cut-off values. HBsAg was detected in 40 oral fluids and 44 whole saliva samples out of 47 paired positive serum specimens and not detected in 64 oral fluids and 63 whole saliva samples out of 68 matched negative sera samples by the ELISA assay. There was excellent agreement between the results for the serum and saliva specimens kappa value (κ): 0.80 for oral fluid and κ: 0.87 for whole saliva and there was excellent reproducibility. Using an optimized protocol, the sensitivities of whole saliva and oral fluid were 93.6 and 85.1%, respectively, whereas specificities of whole saliva and oral fluid were 92.6 and 94.1%, respectively. Our data showed a significant promise for the use of whole saliva and oral fluid together with the modified commercial EIA for Hepatitis B virus infection surveillance.
Asunto(s)
Antígenos de Superficie de la Hepatitis B/análisis , Virus de la Hepatitis B/aislamiento & purificación , Hepatitis B/diagnóstico , Saliva/virología , Adolescente , Adulto , Anciano , Anciano de 80 o más Años , Ensayo de Inmunoadsorción Enzimática , Femenino , Hepatitis B/inmunología , Virus de la Hepatitis B/inmunología , Humanos , Masculino , Persona de Mediana Edad , Reproducibilidad de los Resultados , Adulto JovenRESUMEN
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.
RESUMEN
OBJECTIVES: Little is known about hepatitis A virus (HAV) prevalence in indigenous communities. This study aims to evaluate the prevalence of HAV in indigenous community compared to urban population located at Western Amazon in Brazil. RESULTS: A total of 872 serum samples were obtained from 491 indigenous and 381 non indigenous individuals aging 0 to 90 years. Samples were tested for total and IgM anti-HAV and positive IgM samples were tested for HAV RNA. The overall prevalence of total anti-HAV was 87%, increased according age showing 100% of prevalence in those aging more than 30 years (p < 0.0001) and it was similar among indigenous and urban population. Total anti-HAV prevalence varied between tribes (p < 0.0001) and urban sites (p = 0.0014) and spatial distribution showed high prevalence in homes that received up to 100 dollars. IgM anti-HAV prevalence was 1.7% with predominance in males, those aging more than 41 years. No HAV RNA was detected. In conclusion, high overall anti-HAV prevalence was found in indigenous communities in North Brazil demonstrating the importance of universal vaccination in this group.
Asunto(s)
Hepatitis A , Adulto , Brasil/epidemiología , Hepatitis A/epidemiología , Anticuerpos de Hepatitis A , Humanos , Masculino , Grupos de Población , Prevalencia , Estudios SeroepidemiológicosRESUMEN
Dried blood spots (DBS) testing might increase the access for Hepatitis B virus (HBV) diagnosis, but little is known about the performance of these assays in real life conditions. This study aims to evaluate the diagnostic accuracy of HBsAg, anti-HBc and anti-HBs detection in DBS in clinical settings and field studies and to evaluate demographic and risk behaviour according the presence of HBsAg and anti-HBc. Paired sera and DBS samples were obtained from 2309 individuals from 3 groups, defined as follows: G1: clinical setting (n = 5-19), G2: general population (n = 1305) and G3: vulnerable individuals that could be more exposed to blood contact (n = 485). Sera and DBS were tested using commercial enzyme immunoassay (EIA), with some modifications added. Using DBS samples, the specificity values were above 90 % for HBsAg and anti-HBc in all groups and for anti-HBs range from 58.6%-85%. HBsAg testing had the best performance in GI (sensitivity = 84.4 %) and among those samples that the paired serum also presented anti-HBc marker (sensitivity = 91.6 %). High sensitivity of anti-HBc testing in DBS samples was observed in GI (80.8 %) and among HBV active cases (HBsAg+/anti-HBc+) (98.4 %). Testing of anti-HBs in DBS showed the highest sensitivity in GIII (65.5 %), in previous HBV exposed and cured individuals and when serum titers were above 100 IU/mL (86.7 %). DBS samples could be used for screening and prevalence studies for HBsAg and anti-HBc, particularly in clinical settings and among HBV active cases in field studies.
Asunto(s)
Pruebas con Sangre Seca/normas , Anticuerpos contra la Hepatitis B/sangre , Antígenos de Superficie de la Hepatitis B/sangre , Hepatitis B/diagnóstico , Técnicas para Inmunoenzimas/normas , Adolescente , Adulto , Brasil/epidemiología , Niño , ADN Viral/sangre , Pruebas con Sangre Seca/métodos , Femenino , Hepatitis B/sangre , Hepatitis B/epidemiología , Humanos , Técnicas para Inmunoenzimas/métodos , Masculino , Tamizaje Masivo/métodos , Tamizaje Masivo/normas , Persona de Mediana Edad , Reproducibilidad de los Resultados , Sensibilidad y Especificidad , Adulto JovenRESUMEN
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.
Asunto(s)
Pruebas con Sangre Seca/métodos , Virus de la Hepatitis B/aislamiento & purificación , Hepatitis B/diagnóstico , Biomarcadores/análisis , ADN Viral/análisis , Hepatitis B/virología , Antígenos de Superficie de la Hepatitis B/análisis , Humanos , Sensibilidad y EspecificidadRESUMEN
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.
Asunto(s)
Bioensayo/métodos , Biomarcadores/sangre , Pruebas con Sangre Seca/métodos , Hepatitis B/diagnóstico , Hepatitis C/diagnóstico , Automatización , Humanos , Valor Predictivo de las Pruebas , Sensibilidad y EspecificidadRESUMEN
Few data are available regarding viral hepatitis perception among the general global population. The present study aims to estimate the perception of viral hepatitis in a cohort of individuals living in two geographical regions of Brazil: North (Manaus city (MA)) and Southeast (Rio de Janeiro city (RJ)). A cross-sectional, descriptive study was carried out among 287 subjects recruited in MA (134) and RJ (153). All individuals answered a questionnaire assessing socio-demographic characteristics and viral hepatitis awareness. Participants' responses were scored and divided using interquartile values. Associations between socio-demographic characteristics and knowledge were also evaluated. Interquartile analysis scored 0-21 correct answers as "Very Weak"; 22-27 as "Weak"; 28-31 as "Intermediate"; and 32-47 as "Desirable". Mean ± standard deviations (SD) of correct responses were weak in both MA (24.1 ± 7.0) and RJ (26.3 ± 7.3). Bivariate analysis showed an association between viral hepatitis awareness and both education level (p < 0.001) and family income (p < 0.01). Desirable scores were more common in female participants (61%), those aged between 21-30 years (40%), those with a secondary education (51.7%), those who received high income (31.6%), and those from RJ (70.0%). Health education campaigns in these cities are recommended to increase knowledge and reduce the transmission of these viruses.
Asunto(s)
Conocimientos, Actitudes y Práctica en Salud , Hepatitis Viral Humana , Adolescente , Adulto , Brasil , Ciudades , Estudios Transversales , Escolaridad , Femenino , Humanos , Renta , Masculino , Persona de Mediana Edad , Percepción , Encuestas y Cuestionarios , Adulto JovenRESUMEN
AIM: To evaluate viral hepatitis knowledge among individuals from different resource areas and health conditions to identify possible gaps. METHODS: A cross-sectional, descriptive study was carried out among 447 individuals from five distinct populations in Brazil: Southeast Viral Hepatitis Ambulatory (n = 100), South (n = 89) and Northeast (n = 114) Health Center, Southeast (n = 77) and Northeast (n = 67) low resource areas. All individuals answered a questionnaire assessing sociodemographic characteristics and viral hepatitis awareness. The perception was scored based on the average number of correct answers of all participants and categorized as "low" (0-28 correct answers) or "desirable" (29-46 correct answers). Associations between sociodemographic characteristics and perception were also evaluated. RESULTS: A low level of knowledge was observed in individuals from Northeast Health Center, Northeast and Southeast low resource areas while desirable knowledge was observed in individuals from Viral Hepatitis Ambulatory and South Health Center. According to sociodemographic characteristics, desirable scores were more common among those with secondary education (47.1%), those who declared themselves as white (46.3%), and those who lived in houses with three individuals (25.5%). Multivariate analysis showed an association between viral hepatitis perception and type of population. CONCLUSION: The results demonstrated high level of knowledge among study participants from health clinics from the Southeast region of Brazil and the importance of education programs in increasing the level of knowledge in low resource areas.