Genotyping of cytomegalovirus (CMV) is useful to examine potential differences in the pathogenicity of strains and to demonstrate coinfection with multiple strains involved in CMV disease in adults and congenitally infected newborns. Studies on genotyping of CMV in dried blood spots (DBS) are rare and have been hampered by the small amount of dried blood available. In this study, two multiplex real-time PCR assays for rapid gB and gH genotyping of CMV in DBS were developed. Validation of the assays with 39 CMV-positive plasma samples of transplant recipients and 21 urine specimens of congenitally infected newborns was successful in genotyping 100% of the samples, with gB1 and gB3 being the most prevalent genotypes. Multiple gB and gH genotypes were detected in 36% and 33% of the plasma samples, respectively. One urine sample from a newborn with symptomatic congenital CMV was positive for gB1 and gB2. DBS of congenitally infected newborns (n = 41) were tested using 9 μl of dried blood, and genotypes were detected in 81% (gB) and 73% (gH) of the samples, with gB3 being the most prevalent genotype. No clear association of specific genotypes with clinical outcome was observed. In conclusion, the CMV gB and gH PCR assays were found to be rapid, sensitive for detecting mixed infections, and suitable for direct usage on DBS. These assays are efficient tools for genotyping of CMV in DBS of congenitally infected newborns. Copyright

Cytomegalovirus, accuracy, adolescent, adult, aged, article, bacterial gene, blood sampling, child, clinical article, congenital infection, cytomegalovirus infection, diagnostic test accuracy study, dried blood spot testing, gene targeting, genotype, glycoprotein gB gene, glycoprotein gH gene, graft recipient, hearing loss, human, immunocompromised patient, intermethod comparison, limit of detection, mixed infection, multiplex polymerase chain reaction, newborn, newborn infection, nucleotide sequence, polymerase chain reaction, prevalence, priority journal, real time polymerase chain reaction, school child, sensitivity and specificity, symptomatology, urinalysis, validation process, virus load
dx.doi.org/10.1128/JCM.05253-11, hdl.handle.net/1765/39196
Journal of Clinical Microbiology
Erasmus MC: University Medical Center Rotterdam

de Vries, J.J.C, Wessels, E, Korver, A.M.H, Eijck, A.A, Rusman, L.G, Kroes, A.C.M, & Vossen, A.C.Th.M. (2012). Rapid genotyping of cytomegalovirus in dried blood spots by multiplex real-time PCR assays targeting the envelope glycoprotein gB and gH genes. Journal of Clinical Microbiology, 50(2), 232–237. doi:10.1128/JCM.05253-11