Development of a multiplex PCR for the simultaneous amplification and genotyping of glycoprotein N among human cytomegalovirus strains.
Genomic variation among human cytomegalovirus (HCMV) strains is probably involved in HCMV-induced pathogenesis. The envelope glycoprotein N (gN) showed extensive genetic polymorphism as HCMV isolates have been clustered into four distinct gN variants (gN-1, gN-2, gN-3, gN-4) whose distribution has been analyzed worldwide using different methodological approaches (PCR-RFLP, PCR-Cloning, PCR-Sequencing). This paper describes a new method for concurrent detection of gN genotypes among HCMV strains using a multiplex gN-variants specific PCR plus visualization on agarose gel, avoiding subsequent steps such as cloning, restriction or sequencing. This novel approach will reduce costs and shorten the detection time of gN polymorphisms among HCMV clinical isolates.
Department of Haematology, Oncology and Laboratory Medicine, Clinical Unit of Microbiology, St. Orsola Malpighi General Hospital, University of Bologna, Bologna, Italy. sara.pignatelli@unibo.it
Sara
S
Pignatelli
Daniela
D
Maurizio
Maria Paola
MP
Ladini
Paola
P
Dal Monte
D003587:Cytomegalovirus / Q000145:classification / Q000235:genetics* / Q000302:isolation & purification*; D003586:Cytomegalovirus Infections / Q000821:virology*; D017931:DNA Primers / Q000235:genetics; D005838:Genotype; D006801:Humans; D016133:Polymerase Chain Reaction / Q000379:methods*; D011110:Polymorphism, Genetic; D014759:Viral Envelope Proteins / Q000235:genetics*
D017931:DNA Primers; D014759:Viral Envelope Proteins