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Development of a novel gold nanoparticle-based colorimetric assay for detection of cytomegalovirus (CMV) in pediatric urine

Abstract Summary

Background: One in five infants born with congenital CMV develop complications including hearing loss and developmental delays. Early detection of CMV during the treatable period is critical to prevent long-term complications. Due to the cost-prohibitive nature of testing modalities, most newborns are not tested for CMV if there are no signs of infection. There is a need for a cost-effective, rapid screening method to detect CMV in newborns. Objective: Development of a simple gold nanoparticle (AuNP)-based colorimetric assay for detection of CMV in pediatric urine. Design/Methods: A non-randomized, prospective experimental design with control group. IRB approval was obtained to collect 60 urine samples from pediatric patients at Nemours Children’s Hospital. Confirmatory testing of all urine samples was done by real-time quantitative PCR (qPCR). We tested 16 of the urine samples with our AuNP assay to assess accurate and sensitive CMV detection. Urine was mixed with AuNPs (CytoDiagnostics, Inc.) in the presence or absence of an anti-CMV antibody (Life Technologies). CMV-negative samples were distinguishable from positive samples via an immediately detectable color change from pink to blue. Positive samples appeared pink or purple immediately after mixing. Results: Of the 16 tested urine samples, 7 were CMV-positive by qPCR analysis. Six of the qPCR-positive samples tested positive for CMV in the AuNP assay. There was 1 false negative result. There were 9 samples CMV-negative by qPCR. Of these, 7 samples tested negative for CMV with the AuNP assay. There were 2 false positive results. Compared with qPCR results, the assay has an estimated positive predictive value of 75% and a negative predictive value of 88%. Conclusions: Preliminary results show the AuNP assay was comparable to qPCR analysis. This urine-based detection assay represents an alternative point-of-care test to screen for CMV infection that is a simple, fast, and cost-effective compliment to current gold-standard methods.

Learning Objectives

• Describe the need for a rapid diagnostic test for newborns with congenital CMV.
• Discuss development of a simple, point-of-care diagnostic assay to detect CMV infection in children.
• Compare operator characteristics of the novel nanoparticle-based assay and quantitative PCR for detecting CMV in urine.

Presentation

3440589_16105SonamGupta.pdf

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