Detection and typing of HSV-1, HSV-2, and VZV by a multiplex polymerase chain reaction.
The development of a multiplex polymerase chain reaction method for the rapid and accurate detection and typing of HSV-1, HSV-2, and VZV from clinical specimens is described. A sensitive multiplex polymerase chain reaction was achieved by optimization of parameters such as the primers, magnesium, and dNTPs concentrations. False-negative results that sometimes arise due to inhibitors of DNA amplification or failure of DNA extraction procedure used may be avoided by assaying each specimen with alpha-tubulin primers. Multiplex PCR amplified viral sequences from all 55 specimens obtained from patients with clinical evidence of HSV or VZV infection indicated 100% sensitivity. From 55 patients who were investigated by multiplex PCR, HSV-1 was detected in 28, HSV-2 in 20, and VZV in 7 specimens. The reported results indicate that the present multiplex PCR assay has a potential application in clinical diagnosis when a rapid and accurate detection and typing of involved viruses HSV-1, HSV-2, or VZV is needed.
Markoulatos
P
P
Hellenic Pasteur Institute, Department of Virology, Athens, Greece. vresearch@hol.gr
Georgopoulou
A
A
Kotsovassilis
C
C
Karabogia-Karaphillides
P
P
Spyrou
N
N
D000328:Adult; D018259:Herpesvirus 1, Human / Q000145:classification / Q000302:isolation & purification*; D018258:Herpesvirus 2, Human / Q000145:classification / Q000302:isolation & purification*; D014645:Herpesvirus 3, Human / Q000145:classification / Q000302:isolation & purification*; D006801:Humans; D016133:Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity