Significant compartment‐specific impact of different RNA extraction methods and PCR assays on the sensitivity of hepatitis E virus detection
| Autor: | Eike Steinmann, Patrick Behrendt, Birgit Bremer, Heiner Wedemeyer, Michael P. Manns, Daniel Todt, Markus Cornberg, Benjamin Maasoumy |
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| Přispěvatelé: | CiiM, Zentrum für individualisierte Infektionsmedizin, Feodor-Lynen-Str.7, 30625 Hannover. |
| Rok vydání: | 2021 |
| Předmět: |
ribavirin
Pcr assay medicine.disease_cause Polymerase Chain Reaction Sensitivity and Specificity HEV infection Feces 03 medical and health sciences chemistry.chemical_compound 0302 clinical medicine Hepatitis E virus viral RNA extraction Humans Medicine HEV treatment Detection limit Hepatology business.industry Ribavirin nutritional and metabolic diseases RNA response-guided therapy Virology HEV RNA assay Hepatitis E chemistry 030220 oncology & carcinogenesis Nucleic acid RNA Viral 030211 gastroenterology & hepatology RNA extraction business DNA |
| Zdroj: | Liver international : official journal of the International Association for the Study of the Liver United States |
| ISSN: | 1478-3231 1478-3223 |
| DOI: | 10.1111/liv.14870 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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