Use of stored dried blood spots for retrospective diagnosis of congenital CMV

Atkinson, Claire; Walter, Simone; Tookey, P; Luck, Suzanne; Peckham, Catherine; Griffiths, Paul

doi:10.1002/jmv.21543

Cited by 54 publications

(33 citation statements)

References 13 publications

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“…Detection of CMV DNA in DBS has been shown to be a challenge (3,5,6,9,10,35,36,39) due to the small amount of dried blood (50 l per spot) available. Optimizing the DNA extraction step from the DBS has been shown to result in significantly increased sensitivity of CMV DNA detection in DBS.…”

Section: Discussionmentioning

confidence: 99%

Rapid Genotyping of Cytomegalovirus in Dried Blood Spots by Multiplex Real-Time PCR Assays Targeting the Envelope Glycoprotein gB and gH Genes

et al. 2012

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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 l 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. C ytomegalovirus (CMV) is the most common cause of congenital infection worldwide and an important viral pathogen affecting immunocompromised patients (13, 21). Both in congenitally infected newborns and in immunocompromised patients, genotyping of CMV has been used to study potential differences in pathogenicity of specific strains. However, few authors describe a correlation between specific CMV genotypes and severity of disease (29,32,34,40). More important, genotyping of CMV has enabled the discrimination of reactivation of latent virus from reinfection with new CMV strains in transplant patients, allowing a better definition of donor-to-recipient transmission patterns (24). Congenital CMV infections mainly result from recurrent infections among pregnant women (37), comprising both reactivation and reinfection. The discrimination of reactivation from reinfection may give insight into the mother-to-fetus transmission pattern and the possible associations with the outcome of congenital CMV infections.Genotyping of CMV has mainly focused on envelope glycoproteins gB (UL55) and gH (UL75), which play a role in virus entry and are major targets for neutralizing antibody response. The most frequently used methods for genotyping of CMV are nucleotide sequence analysis (27) and restriction fragment length polymorphism of PCR products (25, 28). Recently, real-time PCR-based assays have been used for rapid detection and quantification of CMV gB and gH genotypes (12,15,24,26). However, they have mainly been applied to plasma or other high-volume samples. Also, deep-sequencing-based methods, sensitive in the de...

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Section: Discussionmentioning

confidence: 99%

Rapid Genotyping of Cytomegalovirus in Dried Blood Spots by Multiplex Real-Time PCR Assays Targeting the Envelope Glycoprotein gB and gH Genes

et al. 2012

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“…Cross-contamination of adjacent stored cards has been reported. 54,[59][60][61] To understand the importance of cCMV as a cause of childhood hearing loss, we reviewed studies that conducted retrospective testing in a group of hearingimpaired children. Requirements were testing by real-time PCR for quantitative analysis of CMV DNA on DBS or on dried umbilical cords.…”

Section: Retrospective Approachmentioning

confidence: 99%

Hearing Loss and Congenital CMV Infection: A Systematic Review

Goderis¹,

Leenheer²,

et al. 2014

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BACKGROUND AND OBJECTIVE: Hearing loss caused by congenital cytomegalovirus (cCMV) infection was first observed in 1964. Today cCMV is the most common cause of nonhereditary sensorineural hearing loss in childhood. Our objective was to provide an overview of the prevalence of cCMV-related hearing loss, to better define the nature of cCMV-associated hearing loss, and to investigate the importance of cCMV infection in hearing-impaired children. METHODS:Two reviewers independently used Medline and manual searches of references from eligible studies and review articles to select cohort studies on children with cCMV infection with audiological follow-up and extracted data on population characteristics and hearing outcomes. RESULTS:Thirty-seven studies were included: 10 population-based natural history studies, 14 longitudinal cohort studies, and 13 retrospective studies. The prevalence of cCMV in developed countries is 0.58% (95% confidence interval, 0.41-0.79). Among these newborns 12.6% (95% confidence interval, 10.2-16.5) will experience hearing loss: 1 out of 3 symptomatic children and 1 out of 10 asymptomatic children. Among symptomatic children, the majority have bilateral loss; among asymptomatic children, unilateral loss predominates. In both groups the hearing loss is mainly severe to profound. Hearing loss can have a delayed onset, and it is unstable, with fluctuations and progression. Among hearing-impaired children, cCMV is the causative agent in 10% to 20%. Despite strict selection criteria, some heterogeneity was found between selected studies.CONCLUSIONS: This systematic review underscores the importance of cCMV as a cause of sensorineural hearing loss in childhood. Pediatrics 2014;134:972-982

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“…T. gondii IgM recovery was significantly impaired in samples stored in ambient conditions (Mei et al, 2011), leading to the exclusion of samples from NC and TX for T. gondii antibody testing. Moreover, CMV DNA viral load is reduced, although still recoverable, after 2 years (Atkinson et al, 2009). Second, previous studies have indicated that neonatal screening for T. gondii IgM using DBS detects ~52-87% of infected newborns (Lebech et al, 1999;Paul et al, 2000;Gilbert et al, 2007).…”

Section: Discussionmentioning

confidence: 98%

A pilot study using residual newborn dried blood spots to assess the potential role of cytomegalovirus and Toxoplasma gondii in the etiology of congenital hydrocephalus

Simeone

Rasmussen

Mei

et al. 2013

Birth Defects Research

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BACKGROUND-Congenital hydrocephalus is a condition characterized by accumulation of cerebrospinal fluid in the ventricles of the brain. Prenatal infections are risk factors for some birth defects. This pilot study investigated whether residual dried blood spots (DBS) could be used to assess infections as risk factors for birth defects by examining the associations between prenatal infection with Toxoplasma gondii (T. gondii) or cytomegalovirus (CMV) with congenital hydrocephalus.

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Use of stored dried blood spots for retrospective diagnosis of congenital CMV

Cited by 54 publications

References 13 publications

Rapid Genotyping of Cytomegalovirus in Dried Blood Spots by Multiplex Real-Time PCR Assays Targeting the Envelope Glycoprotein gB and gH Genes

Rapid Genotyping of Cytomegalovirus in Dried Blood Spots by Multiplex Real-Time PCR Assays Targeting the Envelope Glycoprotein gB and gH Genes

Hearing Loss and Congenital CMV Infection: A Systematic Review

A pilot study using residual newborn dried blood spots to assess the potential role of cytomegalovirus and Toxoplasma gondii in the etiology of congenital hydrocephalus

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