Characterization of Human Adenoviruses of Medical Importance: Isolation of Infectious Virus from Clinical Specimens and Molecular Typing

Abstract

AbstractHuman adenoviruses (HAdVs) constitute a group of ubiquitous viruses that currently comprises 51 well‐defined serotypes and more than 113 genotypes classified into seven species, HAdV‐A through HAdV‐G. The members of these species differ considerably in their genomic characteristics and also in their tropism and pathogenicity. Virus isolation in cell culture remains critical for the preservation and comprehensive characterization of viruses of biomedical interest but has been almost completely abandoned by diagnostic laboratories. Currently, the most frequently used approach for the detection of HAdV in clinical specimens is real‐time qPCR targeting a region of the hexon gene, conserved among all genotypes described to the present. In the absence of typing, the detection of an HAdV in association with disease provides limited information. Molecular typing is therefore highly desirable and required in the epidemiological investigation of HAdV‐associated disease. © 2023 Wiley Periodicals LLC.Basic Protocol 1: Virus isolation from plasma and whole bloodAlternate Protocol 1: Virus isolation from stoolAlternate Protocol 2: Virus isolation from respiratory specimens and urineAlternate Protocol 3: Virus isolation from tissue specimensSupport Protocol: Inoculation of shell vialsBasic Protocol 2: Extraction of highly pure viral genomic DNA from infected cellsBasic Protocol 3: Molecular detection of human adenovirus by real‐time PCRBasic Protocol 4: Molecular typing for basic identification of species and hexon typeBasic Protocol 5: Typing human adenoviruses by next‐generation whole‐genome sequencing and analysis