Rapid detection of herpes simplex virus DNA in genital ulcers by real-time PCR using SYBR green I dye as the detection signal.
We have evaluated a real-time PCR procedure based on the LightCycler technology for rapid detection of herpes simplex virus (HSV) in genital lesions. Two sets of primers, corresponding to the thymidine kinase and DNA polymerase regions, were used for the amplification reactions in separate capillaries containing the SYBR Green I dye as detection signal. In 28 of 118 samples (24%), HSV was isolated by conventional cell culture. All cell culture-positive samples were also positive by real-time PCR. Six additional cell culture-negative samples were positive by PCR with both sets of primers. Total processing time was less than 3 h. Real-time PCR using SYBR Green I as detection signal is a sensitive procedure for the rapid diagnosis of HSV in genital lesions.
Aldea
Carmen
C
Servicio de Microbiología, Hospital 12 de Octubre, Madrid, Spain.
Alvarez
Carmen P
CP
Folgueira
Lola
L
Delgado
Rafael
R
Otero
Joaquín R
JR
D052160:Benzothiazoles; D004279:DNA, Viral / Q000032:analysis*; D003959:Diamines; D005260:Female; D005456:Fluorescent Dyes; D005831:Genital Diseases, Female / Q000821:virology*; D005832:Genital Diseases, Male / Q000821:virology*; D006801:Humans; D008297:Male; D009930:Organic Chemicals; D016133:Polymerase Chain Reaction / Q000379:methods*; D011804:Quinolines; D018139:Simplexvirus / Q000235:genetics / Q000302:isolation & purification*; D014456:Ulcer / Q000821:virology*
D052160:Benzothiazoles; D004279:DNA, Viral; D003959:Diamines; D005456:Fluorescent Dyes; D009930:Organic Chemicals; D011804:Quinolines; C098022:SYBR Green I