title: Genotyping of human cytomegalovirus using non-radioactive single-strand conformation polymorphism (SSCP) analysis.
abstract: Genetic variation in the glycoprotein B (gB) gene may play a role in human cytomegaloviruses (HCMVs) pathogenesis. Using restriction analysis of the gB gene product (PCR-RFLP), amplified by the nested polymerase chain reaction, the HCMV strains can be compared and classified into at least four HCMV groups. PCR single-strand conformation polymorphism (PCR-SSCP) is one of the techniques used to identify a mutant sequence or a polymorphism in a known gene. SSCP analysis has the advantage over RFLP analysis on detection of DNA polymorphisms and point mutations at a variety of positions in DNA fragments. However, the original SSCP protocols using the incorporation of radioactive label and polyacrylamide gel electrophoresis for detection are labour intensive and time-consuming. A simplified SSCP protocol is described to identify HCMV strains and the gB genotype, allowing the detection of sequence variations not residing in the endonuclease recognition sites.
authors.affiliation: Department of Pharmacology, College of Medicine, State University of Campinas, Distrito de Barão Geraldo, SP 13083-970, Campinas, Brazil. dulmal@unicamp.br <dulmal@unicamp.br>
authors.forename: Dulcinéia Martins
authors.initials: DM
authors.lastname: de Albuquerque
authors.forename: Sandra Cecília Botelho
authors.initials: SC
authors.lastname: Costa
mesh_terms: D016026:Bone Marrow Transplantation / Q000009:adverse effects; D003587:Cytomegalovirus / Q000145:classification* / Q000235:genetics*; D003586:Cytomegalovirus Infections / Q000821:virology*; D014644:Genetic Variation; D005838:Genotype; D006801:Humans; D017354:Point Mutation; D016133:Polymerase Chain Reaction; D012150:Polymorphism, Restriction Fragment Length; D018807:Polymorphism, Single-Stranded Conformational; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D014759:Viral Envelope Proteins / Q000235:genetics*
chemical_list: D014759:Viral Envelope Proteins; C040495:glycoprotein B, Simplexvirus