Advertisement

Virology

  • G. J. Boland
Chapter

Abstract

Molecular techniques have become indispensable in viral diagnostics. Current applications include: (1) The detection of (unknown) viral infections in clinical samples. (2) Quantitative monitoring of viral load. (3) Genotyping of viral strains. (4) Detection of mutations in the viral genome that are associated with viral resistance. Proper sample acquisition and sample transport, as well as accurate DNA or RNA isolation are a prerequisite for reliable test results in molecular diagnostics of viral infections. In-house as well as commercial assays can be used for the amplification of viral DNA or RNA for the detection of viral infections and viral load monitoring. Many virus species consist of several subspecies, genotypes or variants. This molecular variation has to be taken into account when applying molecular diagnostics. More complicated diagnostics for genotyping or the detection of mutations related to therapy failure often rely on sequencing, although for some viral targets commercial assays are available. In this chapter, applications are described in which molecular methods have become the most important form of viral diagnostics. Molecular test results have a direct impact on patient management and as such, results have to be reliable, standardized and reproducible. Therefore, quality control and standardization are important issues!

Keywords

Diagnosis Virus detection Variation Genotypes Resistance 

Recommended Literature

  1. Annels NE, et al. Management of Epstein-Barr Virus (EBV) reactivation after allogeneic stem cell transplantation by simultaneous analysis of EBV DNA load and EBV-specific t cell reconstitution. Clin Infect Dis. 2006;42(12):1743–8.CrossRefPubMedGoogle Scholar
  2. Christopherson C, et al. The effects of internal primer-template mismatches on RT-PCR: HIV-1 model studies. Nucleic Acids Res. 1997;25(3):654–8.CrossRefPubMedPubMedCentralGoogle Scholar
  3. Espy MJ, et al. Real-time PCR in clinical microbiology: applications for routine laboratory testing. Clin Microbiol Rev. 2006;19(1):165–256.CrossRefPubMedPubMedCentralGoogle Scholar
  4. Ghedin E, et al. Large-scale sequencing of human influenza reveals the dynamic nature of viral genome evolution. Nature. 2005;437:1162–6.CrossRefPubMedGoogle Scholar
  5. MacKaya WG, et al. Molecular detection and typing of influenza viruses: are we ready for an influenza pandemic? J Clin Virol. 2008;42:194–7.CrossRefGoogle Scholar
  6. Schutten M, et al. Clinical utility of viral quantification as a tool for disease monitoring. Expert Rev Mol Diagn. 2001;1(2):153–62.CrossRefPubMedGoogle Scholar
  7. Smith DB, et al. Expanded classification of hepatitis C virus into 7 genotypes and 67 subtypes: updated criteria and genotype assignment web resource. Hepatology. 2014;59(1):318–27. doi: 10.1002/hep.26744.CrossRefPubMedGoogle Scholar
  8. Stuyver L, et al. Typing of hepatitis C virus isolates and characterization of new subtypes using a line probe assay. J Gen Virol. 1993;74(6):1093–102.CrossRefPubMedGoogle Scholar
  9. Vercauteren J, Vandamme AM. Algorithms for the interpretation of HIV-1 genotypic drug resistance information. Antiviral Res. 2006;71(2–3):335–42.CrossRefPubMedGoogle Scholar
  10. WHO information for molecular diagnosis of influenza virus—update May 2015. Available at: http://www.who.int/influenza/gisrs_laboratory/molecular_diagnosis_influenza_virus_humans_update_201403rev201505.pdf?ua=1. Accessed 28 Jul 2015.

Copyright information

© Springer Nature Singapore Pte Ltd. 2017

Authors and Affiliations

  1. 1.Division Laboratories and Pharmacy, Medical MicrobiologyUMC UtrechtUtrechtThe Netherlands

Personalised recommendations