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Previous issue date: 2015 / Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Rio de Janeiro, RJ, Brasil / O vírus da hepatite E (HEV) é responsável por infecções, em geral, agudas e autolimitantes. No entanto, quando se trata de pacientes imunossuprimidos, a infecção por este vírus pode levar a quadros crônicos e persistentes. Entre os pacientes imunossuprimidos, destacam-se os pacientes HIV positivos \2013 uma população consideravelmente grande, sobre a qual há poucos estudos relacionando a coinfecção HEV/HIV. A hepatite E pode ser causada pelos genótipos 1, 2, 3 e 4 em humanos. O genótipo 3 (HEV-GT3) deve ser destacado por ser tanto o único genótipo circulante relatado no Brasil quanto por ser o que acomete pacientes HIV positivos, levando à cronicidade da doença. Portanto, devido à carência de informações e dados sobre estes pacientes coinfectados e sobre o perfil da hepatite E no Brasil, principalmente devido às dificuldades no diagnóstico, o trabalho buscou aperfeiçoar a técnica de detecção de RT-qPCR e determinar a prevalência da coinfecção HEV/HIV em pacientes do Hospital Universitário Gaffre e Guinle, no Rio de Janeiro para melhor compreensão da coinfecção nesta população
Para tanto, 280 amostras de soro sabidamente positivas para HIV foram coletadas entre os anos de 2012 e 2014, extraídas e testadas por RT-qPCR aperfeiçoado com curva sintética de dsDNA e, posteriormente, com curva sintética de ssRNA, para detecção da ORF3, e controle interno (IPC) para confirmação da coinfecção. As 10 amostras positivas na PCR em tempo real foram testadas em triplicata e por PCR convencional para sequenciamento das regiões das ORFs 1 e 2 e para detecção sorológica de anticorpos anti-HEV IgM e IgG. Porém nenhuma foi positiva para detecção por PCR convencional nem por sorologia, devido a baixa carga viral e ausência de anticorpos anti=HEV IgG e IgM. Nos pacientes em que foi detectado o HEV-RNA, foi observada uma taxa de CD4 e CD8 menores que 1038 e 1254, respectivamente, porém, ainda consideradas normais para indivíduos infectados pelo HIV. A PCR em tempo real foi útil para a detecção de coinfecção HEV/HIV em pacientes com baixa carga viral / The hepatitis E virus (HEV) is generally responsible for acute self
-
limiting infections.
However, when it comes to immunocompromised patients the HEV infecti
on may lead to
chronic and persistent diseases. Among immunosuppressed patients, the HIV positive
patients are highlighted
–
a considerably large population, on which
there are
few studies
relating to H
EV/HIV coinfection.
Hepatitis E can be caused by genot
ypes 1, 2, 3 and 4 in
humans. The genotype 3 of HEV (HEV
-
G
T
3) must be emphasized by being both the only
reported circulating genotype in Brazil and the responsible for chronic disease in HIV positive
patients.
Therefore, due to lack of information and data
on these co
-
infected patients and the
profile of hepatitis E in Brazil, mainly due to difficulties in diagnosis of hepatitis E, the study
aimed at the optimization
of
RT
-
qPCR
detection technique and the determination of HEV/HIV
co
-
infection prevalence on
patients from the Gaffrée & Guinle Hospital, in Rio de Janeiro, for
a better comprehension of this coinfection in this population in Rio de Janeiro city. For that,
280 known HIV positive serum samples were collected between 2012 and 2014, the nucleic
acid
was purified and
tested
by
RT
-
qPCR
technique which was optimized with a synthetic
dsDNA standard curve and, later a synthetic ssRNA standard curve, for detection of HEV
-
RNA ORF3, and internal positive control (IPC) for the co
-
infection confirmation. The 10
HEV/HIV positive samples
for
RT
-
qPCR
were
tested in triplicate and for
qualitative PCR for
ORFs 1 and 2
regions
detection and to antibodies anti
-
HEV IgM and IgG serological
detection, but none was positive for either qualitative PCR or serological tests
d
ue to low viral
titers in serum and lack of antibodies anti
-
HEV IgM and IgG
.
In patients which were positive
for HEV
-
RNA detection, it was observed a CD4 and CD8 rates lower than 1038 and 1254,
respectively
, but still considered normal foi HIV positive
peo
ple
. The real time PCR was
useful for HEV/HIV coinfection detection in patients with low viral rates. / The hepatitis E virus (HEV) is generally responsible for acute self-limiting infections. However, when it comes to immunocompromised patients the HEV infection may lead to chronic and persistent diseases. Among immunosuppressed patients, the HIV positive patients are highlighted \2013 a considerably large population, on which there are few studies relating to HEV/HIV coinfection. Hepatitis E can be caused by genotypes 1, 2, 3 and 4 in humans. The genotype 3 of HEV (HEV-GT3) must be emphasized by being both the only reported circulating genotype in Brazil and the responsible for chronic disease in HIV positive patients. Therefore, due to lack of information and data on these co-infected patients and the profile of hepatitis E in Brazil, mainly due to difficulties in diagnosis of hepatitis E, the study aimed at the optimization of RT-qPCR detection technique and the determination of HEV/HIV co-infection prevalence on patients from the Gaffrée & Guinle Hospital, in Rio de Janeiro, for a better comprehension of this coinfection in this population in Rio de Janeiro city
For that, 280 known HIV positive serum samples were collected between 2012 and 2014, the nucleic acid was purified and tested by RT-qPCR technique which was optimized with a synthetic dsDNA standard curve and, later a synthetic ssRNA standard curve, for detection of HEV-RNA ORF3, and internal positive control (IPC) for the co-infection confirmation. The 10 HEV/HIV positive samples for RT-qPCR were tested in triplicate and for qualitative PCR for ORFs 1 and 2 regions detection and to antibodies anti-HEV IgM and IgG serological detection, but none was positive for either qualitative PCR or serological tests due to low viral titers in serum and lack of antibodies anti-HEV IgM and IgG. In patients which were positive for HEV-RNA detection, it was observed a CD4 and CD8 rates lower than 1038 and 1254, respectively, but still considered normal foi HIV positive people. The real time PCR was useful for HEV/HIV coinfection detection in patients with low viral rates
| Identifer | oai:union.ndltd.org:IBICT/oai:www.arca.fiocruz.br:icict/13811 |
| Date | January 2015 |
| Creators | Lemos, Andreza Salvio |
| Contributors | Moreira, Otacílio da Cruz, Santos, Debora Regina Lopes dos, Fumian, Tulio Machado, Soares, Caroline Cordeiro, Vitral, Claudia Lamarca, Paula, Vanessa Salete de |
| Source Sets | IBICT Brazilian ETDs |
| Language | Portuguese |
| Detected Language | English |
| Type | info:eu-repo/semantics/publishedVersion, info:eu-repo/semantics/masterThesis |
| Source | reponame:Repositório Institucional da FIOCRUZ, instname:Fundação Oswaldo Cruz, instacron:FIOCRUZ |
| Rights | info:eu-repo/semantics/openAccess |
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