A Novel Multiplexed Enzyme-Linked Immunosorbent Assay for the Detection of IgG Seroreactivity to Cytomegalovirus (CMV) UL144
| Autor: | Nathan A. Ledeboer, Ravit Arav-Boger, Matthew L. Faron, Sue C. Kehl, Halli E. Miller, Michael Forman, A Prigan, Pippa Simpson, Brian Mesich |
|---|---|
| Rok vydání: | 2021 |
| Předmět: |
0301 basic medicine
Microbiology (medical) Serotype Human cytomegalovirus Congenital cytomegalovirus infection Cytomegalovirus Enzyme-Linked Immunosorbent Assay Antibodies Viral Serology Viral Proteins 03 medical and health sciences 0302 clinical medicine Antigen Interquartile range Genotype medicine Humans Multiplex 030212 general & internal medicine Child Immunoassays Membrane Glycoproteins business.industry medicine.disease Virology 030104 developmental biology Immunoglobulin G Cytomegalovirus Infections business |
| Zdroj: | J Clin Microbiol |
| ISSN: | 1098-660X 0095-1137 |
| DOI: | 10.1128/jcm.00964-21 |
| Popis: | Infection with human cytomegalovirus (CMV) is common and may have grave consequences in transplant recipients and congenitally infected children. Diagnosis of CMV infection is based on detection of specific antibodies and molecular assays. The incorporation of CMV serological assays into diagnostic algorithms requires careful evaluation and interpretation. Very few serological assays measure CMV infection by a specific strain. We developed an enzyme-linked immunosorbent assay (ELISA) using CMV-encoded UL144 as the antigen. UL144 encodes three major genotypes, A, B, and C, and recombinants. The ELISA was developed with the three UL144 proteins and optimized as a multiplex assay. Sera from 55 positive and 59 negative CMV IgG, determined by the clinical microbiology laboratory, were used for evaluation and optimization. A cutoff optical density (OD) that distinguishes UL144 antibody-positive from antibody-negative sera was established. UL144 A, B, C, and combinations of these antigens were detected in sera. An assay threshold of 0.1 was established and, from a total of 303 sera, the overall sensitivity, specificity, and positive and negative predictive values of the multiplex ELISA were 86.72% (95% confidence interval [CI] 79.59% to 92.07%), 96.57% (92.69% to 98.73%), 94.40% (88.45% to 97.38%), and 91.60% (87.50% to 94.44%), respectively. The inter- and intraassay median coefficients of variation were 0.06 (interquartile range [IQR] 0.56, 0.2) and 0.171 (IQR 0.038, 0.302), respectively. No cross-reactivity was observed with HSV-positive CMV-negative sera. This ELISA gives simple and reproducible results for detection of anti-CMV UL144 IgG. It may assist in differentiating natural infection from CMV vaccines that lack UL144, and may provide an important tool for epidemiological studies of CMV strains. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|