Prospective multicenter comparison of urine culture with PCR on dried blood spots using 2 different extraction and PCR methods in neonates suspected for congenital cytomegalovirus infection

Koen O A Vercauteren; Annelies Keymeulen; Ludo Mahieu; Veerle Cossey; Alexandra Casaer; Christine Van Mol; Koenraad Smets; Elizaveta Padalko

doi:10.1016/j.diagmicrobio.2020.115051

Prospective multicenter comparison of urine culture with PCR on dried blood spots using 2 different extraction and PCR methods in neonates suspected for congenital cytomegalovirus infection

Diagn Microbiol Infect Dis. 2020 Jul;97(3):115051. doi: 10.1016/j.diagmicrobio.2020.115051. Epub 2020 Mar 26.

Authors

Koen O A Vercauteren¹, Annelies Keymeulen², Ludo Mahieu³, Veerle Cossey⁴, Alexandra Casaer⁵, Christine Van Mol⁶, Koenraad Smets², Elizaveta Padalko⁷

Affiliations

¹ Laboratory Medicine Department, Ghent University Hospital, Ghent, Belgium.
² Neonatology Department, Ghent University Hospital, Ghent, Belgium.
³ Neonatology Department, University Hospital Antwerp, University of Antwerp, Belgium.
⁴ Neonatology Department, University Hospital Leuven, Belgium.
⁵ Neonatology Department, AZ St Jan Bruges, Belgium.
⁶ Neonatology Department, ZNA Antwerp, Belgium.
⁷ Laboratory Medicine Department, Ghent University Hospital, Ghent, Belgium. Electronic address: [email protected].

PMID: 32408061
DOI: 10.1016/j.diagmicrobio.2020.115051

Abstract

To evaluate the potential of dried blood spots (DBS) as a congenital cytomegalovirus (cCMV) testing specimen, the laboratory diagnostic accuracy of polymerase chain reaction (PCR) on DBS was compared to viral urine cultures from neonates suspected for cCMV. Two different extraction methods (EasyMAG, bioMérieux versus Qiagen) and 2 real-time PCR protocols (in-house versus Argene) were compared. We were able to collect both DBS and urine samples in 6 Belgian neonatal units from 276 neonates suspected for cCMV registered in CMVREG (an online neonatal registry system). Forty-eight neonates (17.4%) were positive by viral culture in urine. Laboratory diagnostic accuracy parameters of DBS-PCR were both extraction method and PCR protocol dependent. Not all DBS-CMV-PCR methods successfully detected urine-culture-positive neonates born after first-trimester seroconversions. Interestingly, however, all urine-culture-positive neonates having clinical signs of cCMV did consistently score positive.

Keywords: Congenital; Cytomegalovirus; Dried blood spot testing.

Publication types

Comparative Study
Multicenter Study

MeSH terms

Belgium
Cytomegalovirus / genetics
Cytomegalovirus / isolation & purification*
Cytomegalovirus Infections / blood
Cytomegalovirus Infections / congenital*
Cytomegalovirus Infections / diagnosis*
Cytomegalovirus Infections / urine
DNA, Viral / genetics
Dried Blood Spot Testing*
Humans
Infant, Newborn
Prospective Studies
Real-Time Polymerase Chain Reaction* / methods
Sensitivity and Specificity
Urine / virology*
Viral Load

Substances

DNA, Viral