Sequencing and resolution of amplified herpes simplex virus DNA with intermediate melting curves as genotype 1 or 2 by LightCycler PCR assay.
DNA from 101 specimens containing herpes simplex virus (HSV) produced atypical intermediate melting curves compared with those expected for HSV type 1 or HSV type 2 subsequent to real-time PCR. Nucleic acid sequence analysis of amplified target DNA revealed 1- or 3-bp polymorphisms in the probe region which allowed designation of these viruses as HSV genotype 1 or HSV genotype 2. These two subpopulations of HSV were also identified as HSV genotype 1 or HSV genotype 2 using another commercially available PCR method. Amplified HSV target DNA producing intermediate melting curves could be designated as HSV genotype 1 or HSV genotype 2 without performing sequencing or another PCR method with 96/101 (95%) specimens by adding known intermediate HSV DNA characteristic for the two subpopulations as controls.
Issa
Nicolas C
NC
Division of Clinical Microbiology, Mayo Clinic, 200 First St., SW, Rochester, MN 55905, USA.
Espy
Mark J
MJ
Uhl
James R
JR
Smith
Thomas F
TF
D004279:DNA, Viral / Q000032:analysis / Q000737:chemistry; D005838:Genotype; D006558:Herpes Genitalis / Q000821:virology; D006561:Herpes Simplex / Q000821:virology; D018259:Herpesvirus 1, Human / Q000145:classification* / Q000235:genetics; D018258:Herpesvirus 2, Human / Q000145:classification* / Q000235:genetics; D006801:Humans; D016133:Polymerase Chain Reaction / Q000379:methods*; D011110:Polymorphism, Genetic; D012015:Reference Standards; D017422:Sequence Analysis, DNA; D044366:Transition Temperature
D004279:DNA, Viral