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Rapid Detection of Human Cytomegalovirus UL97 and UL54 Mutations Directly from Patient Samples

2013; American Society for Microbiology; Volume: 51; Issue: 7 Linguagem: Inglês

10.1128/jcm.00611-13

1098-660X

Ruth Hall Sedlak, Jared Castor, Susan M. Butler‐Wu, Elaine S. Chan, Linda Cook, Ajit P. Limaye, Keith R. Jerome,

HIV Research and Treatment

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.