Details

Autor(en) / Beteiligte

• 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

Titel

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

Ist Teil von

• Journal of clinical virology, 2009-12, Vol.46, p.S43-S48

Ort / Verlag

Amsterdam: Elsevier B.V

Erscheinungsjahr

2009

Link zum Volltext

Quelle

MEDLINE

Beschreibungen/Notizen

• 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.