Hepatitis E virus infection among patients with altered levels of alanine aminotransferase.

Hepatitis E virus (HEV) affects 20 million people worldwide, with 3.3 million cases and 56,000 deaths. The transmission is mainly by the fecal-oral route. Several studies have reported increased alanine aminotransferase (ALT) levels in association with viral hepatitis. This study evaluated the diagnosis of HEV infection among patients attending the emergency room (ER) of Hospital Beneficência Portuguesa (HBP) and Hospital São Paulo (HSP) in São Paulo, Brazil increased ALT levels (≥ 200 IU/L). From October 2018 to July 2019, 400 sera samples were collected from patients treated at the ER of HBP (n=200) and HSP (n=200). All samples were screened for HEV by RT-qPCR. 200 samples from HSP were tested for IgM of anti-Hepatitis A (HAV) and B (HBV) viruses, and total antibodies of Hepatitis C virus (HCV). Ninety samples (45 from each hospital), were tested for anti-HEV IgM antibodies. Patients aged under 1 to 91 years (mean = 46.29 ± 24.17, median = 48). ALT levels varied from 200 to 8,974 IU/l. 16 patients (4%) turned out positive for HEV by RT-qPCR (ALT levels = 299 to 698 IU/L). Of the 200 HSP patients, 18 (9%) were anti-HAV IgM reactive, 9 (4.5%) for anti-HBV IgM, and 7 (3.5%) for anti-HCV antibodies (ALT levels = 833 to 1918 IU/L). Two of 90 BPH patients (2.22%) were anti-HEV IgM reactive (ALT levels = 1502 to 3831 IU/L). This is the first Brazilian study evaluating patients with suspected HEV infection with increased ALT levels, which were higher than 12 and 60 times the normal upper limit, in the acute phase or for patients reactive for antibody detection, respectively. Liver damage could be minimized by implementing molecular diagnostic tests in the hospital routine.

High prevalence of hepatitis E virus antibodies in Sao Paulo, Southeastern Brazil: analysis of a group of blood donors representative of the general population

Abstract Brazil is a non-endemic country for hepatitis E virus (HEV) infection with seroprevalence from 1% to 4% in blood donors and the general population. However, data on seroprevalence of HEV in the country are still limited. This study evaluated the prevalence of past or present HEV infection in a group of blood donors representative of the general population of the city of Sao Paulo, Southeastern Brazil. Serum samples from 500 blood donors were tested from July to September 2014 by serological and molecular methods. Anti-HEV IgG antibodies were detected in 49 (9.8%) subjects and categorized age groups revealed an age-dependent increase of HEV seroprevalence. Among the anti-HEV IgG positive subjects, only 1 had anti-HEV IgM while none tested positive for HEV-RNA. The present data demonstrate a higher seroprevalence of anti-HEV IgG than previously reported in the region.

High prevalence of hepatitis E virus antibodies in Sao Paulo, Southeastern Brazil: analysis of a group of blood donors representative of the general population.

Brazil is a non-endemic country for hepatitis E virus (HEV) infection with seroprevalence from 1% to 4% in blood donors and the general population. However, data on seroprevalence of HEV in the country are still limited. This study evaluated the prevalence of past or present HEV infection in a group of blood donors representative of the general population of the city of Sao Paulo, Southeastern Brazil. Serum samples from 500 blood donors were tested from July to September 2014 by serological and molecular methods. Anti-HEV IgG antibodies were detected in 49 (9.8%) subjects and categorized age groups revealed an age-dependent increase of HEV seroprevalence. Among the anti-HEV IgG positive subjects, only 1 had anti-HEV IgM while none tested positive for HEV-RNA. The present data demonstrate a higher seroprevalence of anti-HEV IgG than previously reported in the region.

High prevalence of hepatitis E virus antibodies among blood donors in Southern Brazil.

Brazil has been classified as moderately endemic for hepatitis E virus (HEV) infection. However, data on the seroprevalence of HEV in this region are limited. This study evaluated the prevalence of past or present HEV infection among blood donors in the metropolitan area of Itajai Valley, Southern Brazil, a region of predominant German heritage, where cultural habits result in a high consumption of pork. Serum samples from 300 blood donors were tested in December 2014 using serological and molecular methods. Anti-HEV IgG antibodies were detected in 30 (10%) subjects, and categorized age groups revealed an age-dependent increase of HEV seroprevalence. Only one subject had anti-HEV IgM, whereas none tested positive for HEV-RNA. The present data demonstrate a higher seroprevalence of anti-HEV IgG in blood donors than previously reported in Brazil.

Optimisation of a quantitative polymerase chain reaction-based strategy for the detection and quantification of human herpesvirus 6 DNA in patients undergoing allogeneic haematopoietic stem cell transplantation.

Human herpesvirus 6 (HHV-6) may cause severe complications after haematopoietic stem cell transplantation (HSCT). Monitoring this virus and providing precise, rapid and early diagnosis of related clinical diseases, constitute essential measures to improve outcomes. A prospective survey on the incidence and clinical features of HHV-6 infections after HSCT has not yet been conducted in Brazilian patients and the impact of this infection on HSCT outcome remains unclear. A rapid test based on real-time quantitative polymerase chain reaction (qPCR) has been optimised to screen and quantify clinical samples for HHV-6. The detection step was based on reaction with TaqMan® hydrolysis probes. A set of previously described primers and probes have been tested to evaluate efficiency, sensitivity and reproducibility. The target efficiency range was 91.4% with linearity ranging from 10-106 copies/reaction and a limit of detection of five copies/reaction or 250 copies/mL of plasma. The qPCR assay developed in the present study was simple, rapid and sensitive, allowing the detection of a wide range of HHV-6 loads. In conclusion, this test may be useful as a practical tool to help elucidate the clinical relevance of HHV-6 infection and reactivation in different scenarios and to determine the need for surveillance.

Chronic hepatitis E virus infection in a pediatric female liver transplant recipient.

We describe a case of chronic hepatitis E virus (HEV) infection in a 13-year-old female liver transplant recipient with recurrent increased aminotransferase levels and acute cellular rejection. This finding demonstrates that chronic HEV infection can occur and should be further investigated in immunocompromised patients in Latin America.

Short communication: Evaluation of GB virus C/hepatitis G viral load among HIV type 1-coinfected patients in São Paulo, Brazil.

Recent studies suggest that GB virus C/hepatitis G virus (GBV-C/HGV) infection in HIV-positive individuals is associated with a slower progression to AIDS, leading to a lower HIV viral load and higher counts of CD4(+) T cells, although many studies have failed to demonstrate these beneficial effects. We developed a Real-Time PCR (TaqMan RT qPCR) to quantify the viral load of GBV-C/HGV in 102 HIV-1-infected patients, who were also evaluated for the presence of anti-E2. The prevalence of GBV-C/HGV infection was 21% among infected patients and the mean plasma viral load was 3.62 ± 0.64 log(10) copies/ml. Despite the high prevalence, there was no statistical difference when we compared the mean viral load (p≤0.46) and the average count of CD4(+) (p≤0.29) and CD8(+) (p≤0.64) among patients infected by GBV-C/HGV and HIV and patients infected only by HIV. This fact can be explained by the number of patients included in the study. Nevertheless, compared to other studies, we observed a discrete number of patients with undetectable HIV load and lower median viral load in the group presenting GBV-C/HGV RNA. Our study suggests that there may be an impact on HIV viral load in GBV-C/HGV-coinfected patients. However, further studies are needed to elucidate the molecular and cellular mechanisms involved in this viral interaction, previously reported in other studies, with the aim of contributing to the development of new targets for drugs against HIV.

Correlation between specific IgM levels and percentage IgG-class antibody avidity to Toxoplasma gondii.

Toxoplasmosis is an usually asymptomatic worldwide disseminated infection. In its congenital presentation it may lead to abortion or fetal malformations. Antenatal evaluation is considered of paramount importance to identify seronegative women and allow for prophylaxis. Recent improvements in sensitivity of IgM tests has made IgM detection an extremely protracted acute phase marker, and IgG avidity evaluation test became necessary. Observation has shown that a correlation can be established between IgM levels and avidity percentages, suggesting that frequently the avidity test may not be necessary. In this study we analyzed Toxoplasma gondii IgM levels of 202 samples and their IgG avidity percentages, in order to define specific levels whose IgM quantification could by itself define serodiagnosis and therefore make the avidity evaluation unnecessary. We showed that for IgM levels below 2.0 and above 6.0 serodiagnosis of toxoplasmosis could be established without need of IgG avidity test. IgM levels between these two parameters are associated with varying avidity indexes highlighting the importance of its evaluation as a means to confirm toxoplasmosis. Following this demonstration it was possible to avoid the avidity test for 75% of the cases, to reduce the turnaround time and to reduce costs.

Correlation between specific IgM levels and percentage IgG-class antibody avidity to Toxoplasma gondii / Correlação entre níveis de IgM específica e percentual de avidez de anticorpos da classe IgG ao Toxoplasma gondii

Toxoplasmosis is an usually asymptomatic worldwide disseminated infection. In its congenital presentation it may lead to abortion or fetal malformations. Antenatal evaluation is considered of paramount importance to identify seronegative women and allow for prophylaxis. Recent improvements in sensitivity of IgM tests has made IgM detection an extremely protracted acute phase marker, and IgG avidity evaluation test became necessary. Observation has shown that a correlation can be established between IgM levels and avidity percentages, suggesting that frequently the avidity test may not be necessary. In this study we analyzed Toxoplasma gondii IgM levels of 202 samples and their IgG avidity percentages, in order to define specific levels whose IgM quantification could by itself define serodiagnosis and therefore make the avidity evaluation unnecessary. We showed that for IgM levels bellow 2.0 and above 6.0 serodiagnosis of toxoplasmosis could be established without need of IgG avidity test. IgM levels between these two parameters are associated with varying avidity indexes highlighting the importance of its evaluation as a means to confirm toxoplasmosis. Following this demonstration it was possible to avoid the avidity test for 75 percent of the cases, to reduce the turnaround time and to reduce costs.

A Toxoplasmose é uma infecção universal e usualmente assintomática. A forma congênita, entretanto, pode resultar em aborto ou mal formações. Testes sorológicos estão indicados em situações onde há suspeita clínica, e na triagem pré-natal, quando são extremamente importantes para rastrear a infecção e orientar a gestante. O aumento da sensibilidade das técnicas para detecção de IgM, tornou necessário o desenvolvimento de recursos, como a avidez de IgG, visando obter novo marcador de infecção aguda. Embora exista correlação entre níveis de IgM e grau de avidez de IgG, a maioria dos testes de avidez associa-se a níveis baixos de IgM, sugerindo que o teste de avidez não fosse necessário. Portanto, correlacionamos níveis de IgM de 202 amostras com seu respectivo nível de avidez de IgG, dirigidos contra o Toxoplasma, objetivando estabelecer valores claros para a sua indicação. Pôde-se observar que, para IgM < 2,0 e > 6,0, a definição sorológica pode ser feita independentemente da avidez. Níveis de IgM dentro desse intervalo associam-se a índices variados de avidez e, portanto, ressaltam a importância deste teste para definição sorológica do quadro. Com essa abordagem, foi possível diminuir a indicação do teste de avidez em 75 por cento, reduzir o tempo para liberação dos resultados e o custo unitário do teste para IgM.

As novas facetas e a ameaça da gripe aviária no mundo globalizado / New aspects and the threat of an avian flu epidemic in a globalized world

A influenza ou, como é conhecida comumente, a gripe, é uma doença infecciosa aguda causada por um grupo de vírus (com mesmo nome) que acomete várias espécies de animais, desde felinos até aves, passando por humanos. Há cerca de 10 anos têm sido descritos, inicialmente em Hong Kong, surtos de gripe causados por vírus aviários em populações humanas. A seguir, foram descritos surtos na Holanda e no Canadá. Portanto, indaga-se: o mundo corre o risco de um novo surto semelhante ao de 1918, porém com uma população mundial quatro a cinco vezes maior? Qual a velocidade de disseminação desse vírus, visto que as viagens internacionais já não são feitas em navios lentos, mas sim em jatos supersônicos que dão a volta ao mundo em algumas horas? Antes de tudo, o que torna esse vírus tão problemático? Para nós, profissionais de laboratório, é preocupante a questão do diagnóstico etiológico dessas infecções. Como ressaltamos, os quadros de gripe podem ser causados por vários agentes. Portanto, o diagnostico etiológico, básico para intervenções terapêuticas e para que se defina a ocorrência da epidemia, passa a ter relevância ainda maior.

Influenza, or as it is best known, "flu", is an acute respiratory infection caused by a virus that affects many different animal species from felines to birds, including humans.Ten years ago, initialy in Hong Kong and after that in Holand and Canada, outbreaks of avian flu have been increasingly reported in human beings. The question is innevitable: are we in the surge of a new flu pandemic like the one we had in 1918? The perspective is even worse now that we have five times the world population and the international travels are no longer made in slow-going ships but instead in super fast jet planes. For us, laboratory professionals, the question of the laboratory diagnosis is crucial. Considering that so many different viruses are associated with respiratory infections and therapeutic measures depend on this definition, this subject assumes an even greater interest.

The emergence of cytomegalovirus resistance to ganciclovir therapy in kidney transplant recipients.

Transplant recipients that have not been previously exposed to the cytomegalovirus (CMV) are highly susceptible to viral diseases while under immunosuppression therapy. CMV disease requires prolonged therapy, facilitating the emergence of resistant strains. Persistence of positive antigenemia represents clinical evidence of the presence of resistant strains, although its frequency is unknown. These strains may present amino acid deletions or substitutions in conserved regions of the UL97 protein, point mutations in the DNA polymerase (UL54), or both. In this study we aimed to analyze the prevalence of mutations associated with ganciclovir resistance in transplant recipients. Fifteen kidney transplant recipients and four kidney-pancreas transplant recipients, with a positive and oscillating CMV viremia detected by sequential antigenemia test, were enrolled. The UL97 gene was amplified by Nested-PCR and enzymatically digested in samples of these patients in order to detect mutations in the most common codons, such as 460 (M460V), 594 (A594V) and 595(L595S/F). The end-product fragments were further sequenced. Nine (47.4%) out of 19 patients presented with mutations in UL97 at codons L595S (55.6%), A594V (11.1%), A595F/A594V (11.1%) and L595S/A594V (22.2%). None presented with mutation at the M460V codon. Renal transplant patients with oscillation in viral load for more than 2 weeks might have developed viral resistance to anti-drug therapy. Its detection might aid physicians in their clinical plan of tapering the patient's immunosuppression.

Epstein-Barr viral load, interleukin-6 and interleukin-10 levels in post-transplant lymphoproliferative disease: a nested case-control study in a renal transplant cohort.

The possible correlation among Epstein-Barr virus (EBV) load, interleukin-6 (IL-6) and interleukin-10 (IL-10) levels has become an attractive issue and can provide a useful tool for diagnosis and monitoring of patients at risk for post-transplant lymphoproliferative disease (PTLD) development. At the time of diagnosis of PTLD, 11 patients were prospectively enrolled and 55 nested controls were selected from a 1800 renal transplant cohort. Real-time polymerase chain reaction (PCR) was used to quantify EBV load in peripheral blood mononuclear cells (PBMC). Serum IL-6 and IL-10 levels were determined using an enzyme-linked immunosorbent assay (ELISA). The median EBV load of PTLD cases was 17400 copies/10(6) PBMC, statistically different from controls (P=0.001). The median IL-6 level of PTLD cases was not different from controls (P=0.079). However, median IL-10 levels showed a significant difference in both groups (P < or = 0.001). The receiver-operating characteristic (ROC) curve analysis was applied to estimate the IL-10 cut-off value predictive of PTLD development. We found that 73.5 pg/ml has high sensitivity (1.00) and specificity (0.85). Also, Pearson's analysis showed a strong correlation between EBV load and serum IL-10 concentration (P < or = 0.001). This nested case-control study demonstrates that EBV load at diagnosis of PTLD correlates with IL-10 levels, and that monitoring of IL-10 can provide a less expensive and less time-consuming tool for PTLD diagnosis and close follow-up of patients at risk. Furthermore, we were able to define a cut-off value of IL-10 mostly predictive of PTLD development in this cohort. Our data suggest that serial measurements prior to PTLD development must be carried out to validate our hypothesis.

High prevalence of hepatitis C infection in a brazilian prision: identification of risk factors for infection

Hepatitis C virus (HCV) causes infectious hepatitis worldwide. It is transmitted mainly by blood products and sharing of intravenous paraphernalia during illicit drug use. High prevalence rates have been described among specific groups considered to be at higher risk for HCV infection, including prision inmates. The objectives of this study were: to determine the HCV Seroprevalence among inmates of Casa de Detençäo de Säo Paulo; identify risk factors for HCV infection; and to compare the seroprevalence of HCV to other blood borne or sexulally transmitted diseases. From December, 1993, to January, 1994, a total de 779 inmates were interviewed to collect information on sociodemographic status, sexual behavior, and past experience with illict drugs. Blood samples were obtained from 756 inmates for serological tests. 310 (41 percent) blood samples were positive for anti-HCV, 425 (56.2 percent) were negative, and 21 (2.8 percent) showed indeterminate results. In this population, we found a seroprevalence of 13.7 percent for HIV, 3.3 percent for syphylis (VDRL), and 68.1 percent for hepatitis B virus previous infection. Four variables were each identified as associated with a positive anti-HCV serologic test: a positive VDRL (OR = 2.631 IC 95 percent 1.08 to 6.36); a time of current imprisonment longer than 130 months (OR = 2.44 IC 95 percent 1.04 to 5.71); previous incarceration at Casa de Detençäo de Säo Paulo (OR = 1.73 IC 95 percent 1.19 to 2.52) and; illict drug use before admission to the Casa de Detençäo de Säo Paulo (OR = 1.64 IC 95 percent 1.15 to 2.33). The seroprevalence of HCV antibodies among the study populations was high (41 percent), indeed, one of the highest clusters of HCV infection recorded until now. Four variables were each shown to be associated with HCV infection. The simultaneous presence of these 4 analysis indicates most HCV infections occur prior to inprisonment, initiation of control measures to prevent continued transmission after incarceration should be done.

Cytomegalovirus infection in children with Down syndrome in a day-care center in Brazil

This study evaluates the transmission of CMV infection in 120 children aged 1 to 15 years with Down syndrome who attended a day-care center for handicapped children in São Paulo, Brazil. A blood sample was obtained from each children at the beginning of the study for detection of IgG and IgM cytomegalovirus (CMV) antibodies by an immunofluorescence assay. Samples of saliva and urine were obtained every 3 months from the children with CMV antibodies to detect shedding of the virus by culture in human foreskin fibroblasts, by detection of pp65 CMV-antigen and by a nested PCR assay. The prevalence of anti CMV-IgG antibodies was 76.6 per cent (92/120), and IgM anti-CMV antibodies were detected in 13 per cent (12/92) of the seropositive children. During the first viral evaluation, CMV was detected in the urine and/or saliva in 39/90 (43.3 per cent) of the seropositive children. In the second and third evaluations, CMV was detected in 41/89 (46 per cent) and in 35/89 (39.3 per cent) children, respectively. Detection of CMV was shown both in urine and saliva in 28/39 (71.8 per cnet), 19/41(46.3 per cent) and 20/35 (57.1 per cent) of the children excreting the virus, respectively...