EVALUATION OF RESTRICTION-FRAGMENT-LENGTH-POLYMORPHISM ANALYSIS OF THE UL10-UL13 GENOMIC REGION FOR RAPID IDENTIFICATION OF HUMAN CYTOMEGALOVIRUS STRAINS

A sensitive semi-nested polymerase chain reaction (PCR) was establishe d which allows rapid identification of human cytomegalovirus strains d irectly on clinical specimens, thereby permitting virus isolation and propagation on cell cultures to be avoided. The assay is based on rest riction analysis of PCR products derived from the polymorphic UL10-UL1 3 region of the human cytomegalovirus genome. The method was evaluated using clinical samples from 23 subjects comprising 16 breast-feeding mothers and seven bone marrow transplantation recipients. For eight mo thers, postnatal virus transmission to their offspring via breast milk was studied. Interestingly, for one mother-infant pair, a double infe ction with two distinct human cytomegalovirus strains could be demonst rated. Stepwise digestion with different restriction enzymes raised th e possibility of detecting different strains almost twofold compared t o analysis with only one enzyme. This assay is a practical tool for mo nitoring human cytomegalovirus transmission in various clinical settin gs.