Open AccessFailure To Genotype Herpes Simplex Virus by Real-Time PCR Assay and Melting Curve Analysis Due to Sequence Variation within Probe Binding Sites
Author(s) -
Trevor P. Anderson,
Anja Werno,
Kirsten A. Bey,
David R. Murdoch
Publication year - 2003
Publication title -
journal of clinical microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.349
H-Index - 255
eISSN - 1070-633X
pISSN - 0095-1137
DOI - 10.1128/jcm.41.5.2135-2137.2003
Subject(s) - melting curve analysis , genotyping , herpes simplex virus , biology , virology , genotype , polymerase chain reaction , real time polymerase chain reaction , sequence analysis , microbiology and biotechnology , dna , virus , gene , genetics
Real-time PCR with melting curve analysis of PCR products is a rapid procedure for detecting and genotyping herpes simplex virus (HSV). When testing mucocutaneous samples for HSV by a real-time PCR assay targeting the DNA polymerase gene, we found that some PCR products had atypical melting curves that did not conform to the expected melting temperatures for HSV type 1 or 2. Sequence analysis showed that these strains had base-pair mismatches over the probe binding sites. An alternative assay is required to type such atypical isolates.