RESUMO
BACKGROUND: Hepatitis E virus (HEV) is an increasing cause of acute viral hepatitis in developed countries. Serological testing alone may fail to diagnose acute infection, especially in immunocompromised patients, which justifies the use of molecular assays for diagnosis. Few studies have compared accuracy of HEV RNA detection assays. OBJECTIVES: The performances of five real-time PCR procedures for HEV RNA detection were compared. STUDY DESIGN: First, RNA quantification of hepatitis E diluted standards of 3a and 3b genotypes were performed. Secondly, forty-seven clinical samples of patients with known acute HEV infection were tested using five hepatitis E RNA detection methods of assigned letters A, B, C, D and E. RESULTS: Standards of HEV 3a genotype were detected in 100% of replicates with 2500 UI/ml of sensitivity by using A, B and C assays. Standards of HEV 3b genotype were more accurately detected with a sensitivity of 25 UI/ml in 100% of replicates using C assay and were detected in 100% of replicates with 2500 UI/ml of sensitivity by using A, B and E assays. Overall, B assay detected all of 250 UI/ml dilution and occasionally the 25 UI/ml dilution on both subtypes. The detection rates of clinical samples were 100%, 100% 97%, 97% and 83% for the respective A, B, C, D and E assay. Assays A and B were well correlated, independently of the subtype. However, discrepancies were observed when these techniques were compared to C, D and E assays according to the different subtypes. CONCLUSION: A and B assays appear reliable for HEV RNA detection. These assays target the ORF2/3 overlapping region, described as more conserved than ORF2.
Assuntos
Vírus da Hepatite E/isolamento & purificação , Hepatite E/diagnóstico , Técnicas de Diagnóstico Molecular/métodos , RNA Viral/isolamento & purificação , Reação em Cadeia da Polimerase em Tempo Real/métodos , Virologia/métodos , Hepatite E/virologia , Vírus da Hepatite E/genética , Humanos , RNA Viral/genética , Sensibilidade e EspecificidadeRESUMO
Among hepatitis E virus (HEV) infections diagnosed in 2011 by the French Reference Centre for HEV, 9 were due to genotype 4, which until recently was limited to Asia. Sequences from autochthonous cases formed a single cluster very similar to Belgian swine sequences. Clinical presentation differed from genotype 3 infections.
Assuntos
Vírus da Hepatite E/classificação , Vírus da Hepatite E/genética , Hepatite E/epidemiologia , Hepatite E/patologia , Adulto , Idoso , Análise por Conglomerados , Feminino , França/epidemiologia , Genótipo , Hepatite E/virologia , Vírus da Hepatite E/isolamento & purificação , Humanos , Masculino , Pessoa de Meia-Idade , Epidemiologia Molecular , Dados de Sequência Molecular , Filogenia , RNA Viral/genética , Análise de Sequência de DNARESUMO
Hepatitis E is currently diagnosed after all other causes of hepatitis have been excluded. Moreover, HEV testing is not performed to prevent blood transmission in developed countries. We report here on the case of a patient with acute hepatitis while receiving potentially hepatotoxic medications for autoimmune disorders, with low-level autoimmune markers and negative "standard" viral markers; it was finally determined that he was suffering from transfusion-transmitted hepatitis E.
