Characterization of a universal screening approach for congenital CMV infection based on a highly-sensitive, quantitative, multiplex real-time PCR assay


Autoři: Angela Nagel aff001;  Emmanouela Dimitrakopoulou aff002;  Norbert Teig aff003;  Peter Kern aff004;  Thomas Lücke aff003;  Dariusz Michna aff005;  Klaus Korn aff001;  Philipp Steininger aff001;  Khalid Shahada aff006;  Katrin Neumann aff002;  Klaus Überla aff001
Působiště autorů: Institute of Clinical and Molecular Virology, University Hospital Erlangen, Erlangen, Germany aff001;  Department of Otorhinolaryngology, Head and Neck Surgery, Division of Phoniatrics and Pediatric Audiology, St. Elisabeth-Hospital, Ruhr University Bochum, Bochum, Germany aff002;  Department of Pediatrics, St. Josef-Hospital, Ruhr University Bochum, Bochum, Germany aff003;  Department of Gynecology, St. Elisabeth-Hospital, Ruhr University Bochum, Bochum, Germany aff004;  Department of Pediatrics, Elisabeth-Hospital Essen, Germany aff005;  Audiology and Balance Center, Hamad Medical Corporation, Doha, Qatar aff006
Vyšlo v časopise: PLoS ONE 15(1)
Kategorie: Research Article
doi: 10.1371/journal.pone.0227143

Souhrn

The majority of congenital cytomegalovirus (cCMV) infections are asymptomatic at birth and therefore not diagnosed. Approximately 10–15% of these infants develop late-onset hearing loss and other developmental disorders. Implementation of a universal screening approach at birth may allow early initiation of symptomatic interventions due to a closer follow-up of infants at risk and offers the opportunity to consider treatment of late-onset disease. Real-time PCR assays for the detection of CMV DNA in buccal swab samples demonstrated feasibility and good clinical sensitivity in comparison to a rapid culture screening assay. Because most cCMV infections remain asymptomatic, a universal screening assay that stratifies CMV infected infants according to low and high risk of late-onset cCMV disease could limit the parental anxiety and reduce follow-up costs. We therefore developed and characterized a screening algorithm based on a highly-sensitive quantitative real-time PCR assay that is compatible with centralized testing of samples from universal screening and allows to determine CMV DNA load of saliva samples either as International Units (IU)/ml saliva or IU/105 cell equivalents. 18 of 34 saliva samples of newborns that tested positively by the screening algorithm were confirmed by detection of CMV DNA in blood and/or urine samples obtained during the first weeks of life. All screening samples that could not be confirmed had viral loads of <2.3x105 IU/ml saliva (median: 6.8x103) or 1.3x105 IU/105 cell equivalents (median: 4.0x102). The viral load of screening samples with confirmed cCMV infection ranged from 7.5x102 to 8.2x109 IU/ml saliva (median: 9.3x107) or 1.5x102 to 5.6x1010 IU/105 cell equivalents (median: 3.5x106). Clinical follow-up of these newborns with confirmed cCMV infection should reveal whether the risk of late-onset cCMV disease correlates with CMV DNA load in early life saliva samples and whether a cut-off can be defined identifying cCMV infected infants with or without risk for late-onset cCMV disease.

Klíčová slova:

Albumins – Blood – DNA extraction – Neonates – Polymerase chain reaction – Saliva – Urine – Viral load


Zdroje

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Článek vyšel v časopise

PLOS One


2020 Číslo 1