Rapid, differential diagnosis of orthopox- and herpesviruses based upon real-time PCR product melting temperature and restriction enzyme analysis of amplicons.
Orthopoxviruses tend to have non-specific early symptoms that cannot be differentiated readily from other infectious exanthemas, such as varicella-zoster virus (VZV) or disseminated herpes simplex virus (HSV) infections. A rapid assay was developed for the differential diagnosis of orthopoxviruses and herpesviruses based upon the melting temperatures of real-time PCR amplicons. A mean melting temperature difference of 8.7 degrees C was observed between the products amplified from the two virus families. Further identification of individual pathogens was made using restriction enzyme analysis. The assay was able to identify correctly viruses from quality control panels of herpes and orthopoxviruses.
Carletti
Fabrizio
F
Laboratory of Virology, National Institute for Infectious Diseases Lazzaro Spallanzani - IRCCS, Via Portuense, 292, 00149 Rome, Italy.
Di Caro
Antonino
A
Calcaterra
Silvia
S
Grolla
Allen
A
Czub
Markus
M
Ippolito
Giuseppe
G
Capobianchi
Maria R
MR
Horejsh
Douglas
D
D017931:DNA Primers; D006561:Herpes Simplex / Q000175:diagnosis* / Q000821:virology; D006564:Herpesviridae / Q000235:genetics / Q000302:isolation & purification*; D006801:Humans; D018155:Orthopoxvirus / Q000235:genetics / Q000302:isolation & purification*; D016133:Polymerase Chain Reaction / Q000379:methods*; D011213:Poxviridae Infections / Q000175:diagnosis*; D015183:Restriction Mapping / Q000379:methods*; D012680:Sensitivity and Specificity; D013696:Temperature
D017931:DNA Primers