Direct genotyping of cytomegalovirus envelope glycoproteins from toddler's saliva samples

Abstract Background The polymorphism of genes encoding CMV envelope protein is used for strain classification and may influence pathogenesis and/or infectivity. CMV genotyping is usually based on sequencing or acrylamide gel-RFLP, but these methods are not suited to rapid screening of large populati...

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Bibliographic Details
Published in:Journal of clinical virology 2009-12, Vol.46, p.S43-S48
Main Authors: Grosjean, J, Hantz, S, Cotin, S, Baclet, M.C, Mengelle, C, Trapes, L, Virey, B, Undreiner, F, Brosset, P, Pasquier, C, Denis, F, Alain, S
Format: Article
Language:English
Subjects:
Allergy and Immunology
Biological and medical sciences
Child Day Care Centers
Cytomegalovirus
Cytomegalovirus - classification
Cytomegalovirus - genetics
Cytomegalovirus - isolation & purification
Cytomegalovirus Infections - diagnosis
Cytomegalovirus Infections - virology
Fundamental and applied biological sciences. Psychology
Genotype
Glycoproteins
Human viral diseases
Humans
Infant
Infectious Disease
Infectious diseases
Medical sciences
Microbiology
Miscellaneous
PCR-RFLP
Polymerase Chain Reaction - methods
Polymorphism, Restriction Fragment Length
Polymorphism, Single Nucleotide
Saliva
Saliva - virology
Techniques used in virology
Toddler
Viral diseases
Viral Envelope Proteins - genetics
Viral Proteins - genetics
Virology
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Summary:Abstract Background The polymorphism of genes encoding CMV envelope protein is used for strain classification and may influence pathogenesis and/or infectivity. CMV genotyping is usually based on sequencing or acrylamide gel-RFLP, but these methods are not suited to rapid screening of large populations. Objectives We developed a high-throughput method to analyze CMV strains diversity and to detect multiple-strain infection in a large population of toddlers (six daycare centers (DCC) and an emergency unit (EU)). Methods We developed a new PCR-RFLP method coupled with capillary electrophoresis fragment detection for UL55 -gB, UL75 -gH and UL73 -gN genotyping. To detect gB recombinants, gpUL55 typing was applied to two variable zones (NTerminal and central). We applied this method to 212 CMV-positive saliva samples and controlled the results by direct sequencing of PCR products. Results We identified 112 strains, that fell into eight groups in UL55-gB, two groups in UL75-gH, and seven groups in UL73-gN. The 79 samples from the emergency unit contained 30 strains, 28 children harboring 2 strains. The samples ( n = 133) from the six daycare centers contained respectively 4, 1, 6, 1 and 11 strains. Fifteen percent of strains were UL55 -gB recombinants. Conclusion Our new method can simultaneously determine gB, gH and gN genotypes and offers more precise classification of CMV strains than previous RFLP-based methods. This could constitute the basis for a new classification, particularly in UL55 -gB. Easy direct identification of multiple strains and recombinants in pathological samples could facilitate large epidemiologic studies.
ISSN:1386-6532
1873-5967
DOI:10.1016/j.jcv.2009.08.018