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Significant compartment-specific impact of different RNA extraction methods and PCR assays on the sensitivity of hepatitis E virus detection.
Behrendt, Patrick; Bremer, Birgit; Todt, Daniel; Steinmann, Eike; Manns, Michael Peter; Cornberg, Markus; Wedemeyer, Heiner; Maasoumy, Benjamin.
Afiliación
  • Behrendt P; Department of Gastroenterology, Hepatology and Endocrinology, Hannover Medical School, Hannover, Germany.
  • Bremer B; Institute of Experimental Virology, TWINCORE, Centre for Experimental and Clinical Infection Research, A Joint Venture Between the Medical School Hannover (MHH), and the Helmholtz Centre for Infection Research (HZI, ), Hannover, Germany.
  • Todt D; German Centre for Infection Research (DZIF), Partner-site Hannover-Braunschweig, Germany.
  • Steinmann E; Department of Gastroenterology, Hepatology and Endocrinology, Hannover Medical School, Hannover, Germany.
  • Manns MP; Department of Molecular and Medical Virology, Ruhr-University Bochum, Bochum, Germany.
  • Cornberg M; European Virus Bioinformatics Center (EVBC), Jena, Germany.
  • Wedemeyer H; Department of Molecular and Medical Virology, Ruhr-University Bochum, Bochum, Germany.
  • Maasoumy B; Department of Gastroenterology, Hepatology and Endocrinology, Hannover Medical School, Hannover, Germany.
Liver Int ; 41(8): 1815-1823, 2021 08.
Article en En | MEDLINE | ID: mdl-33683813
BACKGROUND: RNA detection in plasma/stool is the gold-standard for diagnosis of hepatitis E virus (HEV) infection. The impact of viral extraction methods on HEV RNA detection is poorly investigated. METHODS: We determined the limit of detection of the RealStar HEV RT-PCR V2.0 Kit (altona Diagnostics, RS) utilizing 3 RNA extraction methods (COBAS® AmpliPrep Total Nucleic Acid Isolation Kit, TNAi Roche; MagNA Pure 96 DNA, Viral NA SV Kit, MgP; QIAamp Viral RNA mini Kit Qiagen; VRK) in plasma and stool. The most sensitive method was evaluated in a total of 307 longitudinal samples of patients with HEV infection (acute = 18/chronic = 36) and compared to results with the former diagnostic standard of our centre (TNAi/FastTrack Diagnostic; FTD). RESULTS: The plasma-LOD was 49, 94 and 329 IU/mL for extraction with MgP, VRK and TNAi respectively. In stool, the LOD was 21 IU/mL, 528 IU/mL and indefinable for extraction with TNAi, VRK and MgP respectively. Utilizing longitudinal patient plasma samples, MgP/RS revealed 56 HEV RNA-positive samples in 158 negative samples as determined by TNAi/FTD. In stool, from 37 HEV negative samples (TNAi/FTD), 15 were positive with TNAi/RS. At end of treatment, 8 out of 27 chronically infected patients were RNA positive with MgP/RS, while classified negative with TNAi/FTD. A relapse occurred in 3 of these patients. CONCLUSION: Different methods for RNA extraction and quantification have a significant, compartment-specific impact on the sensitivity of HEV detection. Knowledge about the favourable combinations of extraction and quantification has important implications for diagnosis and patients receiving antiviral therapy.
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Virus de la Hepatitis E / Hepatitis E Tipo de estudio: Diagnostic_studies Límite: Humans Idioma: En Revista: Liver Int Asunto de la revista: GASTROENTEROLOGIA Año: 2021 Tipo del documento: Article País de afiliación: Alemania Pais de publicación: Estados Unidos
Texto completo
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Imprimir
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Virus de la Hepatitis E / Hepatitis E Tipo de estudio: Diagnostic_studies Límite: Humans Idioma: En Revista: Liver Int Asunto de la revista: GASTROENTEROLOGIA Año: 2021 Tipo del documento: Article País de afiliación: Alemania Pais de publicación: Estados Unidos