V.S. de Paula

Seroprevalence of viral hepatitis in riverine communities from the Western Region of the Brazilian Amazon Basin

The western region of the Brazilian Amazon Basin has long been shown to be a highly endemic area for hepatitis B and hepatitis D viruses. Data concerning the prevalence of hepatitis C and E viruses in this region are still scarce. In this study we investigated the presence of hepatitis A, B, C, D and E viruses infection in communities that live along the Purus and Acre rivers in the states of Acre and Amazonas within the Amazon Basin. A total of 349 blood samples were collected and tested for hepatitis A-E serological markers (antibodies and/or antigens) using commercial enzyme linked immunosorbent assays. Anti-HCV positive sera were further assayed by an immunoblot. HBsAg positive sera were subtyped by immunodifusion. The overall prevalence for hepatitis A, B, C, and E were 93.7%, 66.1%, 1.7%, and 4%, respectively. A very high prevalence of delta hepatitis (66.6%) was found among HBsAg positive subjects. Hepatitis A, B and D viruses were shown to be largely disseminated in this population, while hepatitis C and E viruses infection presented low prevalence rates in this region. The analysis of risk factors for HBV infection demonstrated that transmission was closely associated with sexual activity.

Molecular detection of hepatitis A virus in urban sewage in Rio de Janeiro, Brazil.

Aims:  A one‐year survey was conducted to examine hepatitis A virus (HAV) prevalence, distribution of genotypes and their relationship to bacterial indicators in raw and treated sewage samples.

High prevalence of human Torque teno virus in streams crossing the city of Manaus, Brazilian Amazon

Aims:  Torque teno virus (TTV) is a human DNA virus chronically infecting most healthy individuals worldwide and can be transmitted by faecal–oral route. The occurrence of TTV was evaluated in the streams crossing the city of Manaus (Brazilian Amazon) over a 1‐year period, four times a year.

Detection of hepatitis A virus RNA in serum during the window period of infection

BACKGROUND Hepatitis A virus (HAV) infection is the leading cause of clinically apparent viral hepatitis in many parts of the world, including developed and developing countries. Only limited information is available regarding the seronegative viremic window that follows HAV infection, and no systematic search has been reported for HAV RNA positive, IgM anti-HAV negative serum samples during hepatitis A outbreaks. OBJECTIVES To determine the proportion of HAV infected individuals among (i) children who were tested negative for anti-HAV antibodies during hepatitis A outbreaks which occurred in a public school (n = 157) and a child care center (n = 38); (ii) subjects (n = 46) initially classified as acute non-A-C hepatitis patients after clinical examination and serological tests (sporadic cases). STUDY DESIGN Reverse transcription (RT)-PCR was performed to detect the presence of HAV genome in serum samples collected from anti-HAV negative, susceptible subjects. RESULTS HAV RNA was detected in 19/157 (12%) and 5/38 (13%) anti-HAV negative children from the public school and child care center, respectively. Twelve (26%) out of the 46 acute hepatitis patients (sporadic cases) were also HAV RNA positive. From nine of these 12 patients, a second blood sample was obtained 18-34 days after the first one: all nine had seroconverted to IgM anti-HAV, and their serum transaminases had reached elevated levels (mean ALT, 418; mean AST, 241). CONCLUSIONS Detection of HAV RNA before IgM anti-HAV seroconversion may be used as an early diagnosis method during hepatitis A outbreaks. HAV RNA testing should also help to elucidate acute hepatitis cases of unknown etiology.

Hepatitis A virus infection in hepatitis C Brazilian patients

OBJECTIVE HAV infection in patients with pre-existing chronic liver disease has been associated with increased rate of fulminant hepatitis and mortality. The aim of this study was to investigate the presence of serological and molecular HAV markers in a population of HCV infected patients. PATIENTS AND METHODS The presence of total and IgM anti-HAV antibodies was investigated in 197 patients (mean age 44.8+/-12.5 years) referred to the Brazilian Reference Center for Viral Hepatitis and who tested positive for anti-HCV antibodies and HCV RNA. HAV RNA was investigated by reverse transcription-nested PCR in these patients.Results. One hundred seventy patients (86%) had total, but not IgM anti-HAV antibodies, being therefore, immune to hepatitis A, while 27 (14%) were not. A high proportion (6/27, 22%) of the susceptible patients presented markers of recent HAV infection: One patient was IgM anti-HAV positive, three were HAV RNA positive, and two presented both markers. By nucleotide sequencing, it was demonstrated that the HAV isolates infecting these patients belonged to subgenotypes 1A and 1B. CONCLUSIONS Superinfection with HAV was a common event in the group of HCV infected patients under study. Implementation of hepatitis A vaccination should be considered for this population.

Kinetics of hepatitis A virus replication in vivo and in vitro using negative-strand quantitative PCR

The replication of hepatitis A virus (HAV) is via a complementary negative-strand RNA. Each negative strand may serve as a template for the synthesis of many positive strands. The aim of this study was to detect the intermediate replicative (negative strand) of HAV in order to monitor its replication in vitro and in vivo. Real-time polymerase chain reaction (PCR) was standardized to detect the intermediate replicative of HAV in cell culture and liver from non-human primates infected experimentally. HAV primers from the 5′ non-translated region and VP3 were used in the cDNA synthesis of negative-strand RNA. The negative strand was detected in the infected cell lines and liver by highly strand-specific rTth recombinant Thermus thermophilus DNA polymerase reverse transcription followed by quantitative PCR. The results indicate that the negative-strand HAV RNA can be detected in vivo and in vitro. This model is an approach for assessing the dynamic patterns of replication and should represent a valuable tool for the monitoring of HAV replications in cell cultures and for the evaluation of experimental infections in animal models.

Changes in hepatitis A virus seroepidemiology in HIV-infected Brazilian patients

Shifting of hepatitis A virus (HAV) epidemiology from a high towards an intermediate endemicity pattern and use of antiretroviral therapy increased the risk of HIV/HAV coinfection in developing countries. The aim of this study was to investigate the presence of HAV markers in a cohort of HIV-infected patients from 1988 to 2004. The presence of serum anti-HAV antibodies and HAV-RNA by real-time polymerase chain reaction was investigated in 581 patients. Total anti-HAV antibodies was found in 464/581 (79.8%) patients, however, achanging epidemiologic pattern of hepatitis A among HIV-infected patients from 1988 to 2004 was observed. Among patients susceptible to HAV (n = 117), 5 (4.2%) were co-infected with HAV, all of them had IgM anti-HAV antibodies and were serum HAV-RNA-positive. The high prevalence of anti-HAV antibodies in HIV-infected patients suggests that screening tests for anti-HAV antibodies should be performed before implementation of hepatitis A vaccination, especially in those patients from endemic countries.

Detection and quantification of hepatitis E virus in the absence of IgG and IgM anti-HEV in HIV-positive patients

To improve RT‐qPCR with an internal control and a synthetic standard curve to detect HEV in HIV co‐infected patients.

LB1.63 High acceptance of cervical self-collection for detection of hpv, hhv-2 and hiv-1 in women living in tapajÓs region, amazÔnia, brazil

Introduction Cervical self-collection is a safe and efficient method for detecting sexually transmitted infections (STIs). The study aims to verify the acceptance of cervical self-collection and the prevalence of HPV, HHV-2 and HIV-1 infection in women living in the Tapajós region, Amazônia, Brazil. Methods Cross-sectional study with women attending in Santarém-Pará. The collection was performed between August 2015 and January 2017. Participants collected cervical scrapings and peripheral blood. Those who accepted, also performed cervical self-collection. Detection of HPV DNA was performed by nestedPCR with MY09/11 and GP5/6+ primers and typing was done by sequencing. Detection of HHV-2 DNA was performed by real-time PCR with Taqman. Identification of anti-HIV-1/2 antibodies was made by Alere Determine Kit. Results A total of 206 specimens were obtained from 112 women. The acceptance of cervical self-collection was 84% (94/112) and HPV DNA was identified in 39.4% (37/94) of the samples. While the prevalence of HPV infection in cervical scraping was 32.1% (36/112). All the women presented Papanicolaou negative for malignancy. The most prevalent types were HPV-16 and HPV-18. The overall prevalence of HHV-2 infection was 8.9%. The concordance rate in the molecular diagnosis between cervical scraping and cervical self-collection was 65% (26/40) for HPV and 50% (4/8) for HHV-2. No woman had HIV-1 reactive serology. Conclusion A high prevalence of HPV infection was found in women without dysplastic lesion. Cervical self-collection had high acceptance, moderate concordance rate in the detection of HPV DNA compared to cervical scraping, and alone it was more efficient in the detection of HPV. This is the first study in women living in Tapajós region and the findings strongly suggest that cervical self-collection may be a useful tool for increasing access to diagnosis of STIs and screening for cervical cancer in women living in the Amazon.