title: Rapid detection of human cytomegalovirus UL97 and UL54 mutations directly from patient samples.
abstract: Human cytomegalovirus (CMV) is a significant contributor to morbidity and mortality in immunocompromised patients, particularly in the transplant setting. The availability of anti-CMV drugs has improved treatment, but drug resistance is an emerging problem. Here, we describe an improved, rapid, sequencing-based assay for the two genes in CMV where drug resistance occurs, the UL97 and UL54 genes. This assay is performed in 96-well format with a single master mix and provides clinical results within 2 days. It sequences codons 440 to 645 in the UL97 gene and codons 255 to 1028 in the UL54 gene with a limit of detection of 240 IU/ml. With this assay, we tested 43 specimens that had previously been tested for UL97 drug resistance and identified 3 with UL54 mutations. One of these patients had no concurrent UL97 mutation, pointing toward the need for an assay that facilitates dual UL97/UL54 gene testing for complete resistance profiling.
authors.affiliation: Department of Laboratory Medicine, University of Washington Medical Center, Seattle, Washington, USA. rah7@u.washington.edu
authors.forename: Ruth
authors.initials: R
authors.lastname: Hall Sedlak
authors.forename: Jared
authors.initials: J
authors.lastname: Castor
authors.forename: Susan M
authors.initials: SM
authors.lastname: Butler-Wu
authors.forename: Elaine
authors.initials: E
authors.lastname: Chan
authors.forename: Linda
authors.initials: L
authors.lastname: Cook
authors.forename: Ajit P
authors.initials: AP
authors.lastname: Limaye
authors.forename: Keith R
authors.initials: KR
authors.lastname: Jerome
mesh_terms: D003587:Cytomegalovirus / Q000235:genetics* / Q000302:isolation & purification; D003586:Cytomegalovirus Infections / Q000821:virology*; D004259:DNA-Directed DNA Polymerase / Q000235:genetics*; D024882:Drug Resistance, Viral; D006801:Humans; D025202:Molecular Diagnostic Techniques / Q000379:methods; D050505:Mutant Proteins / Q000235:genetics; D009154:Mutation; D017853:Phosphotransferases (Alcohol Group Acceptor) / Q000235:genetics*; D014764:Viral Proteins / Q000235:genetics*; D014773:Virology / Q000379:methods*
chemical_list: D050505:Mutant Proteins; C109367:UL54 protein, Human herpesvirus 5; D014764:Viral Proteins; D017853:Phosphotransferases (Alcohol Group Acceptor); C075365:ganciclovir kinase; D004259:DNA-Directed DNA Polymerase