Quantitation of HHV-7 genome by real-time polymerase chain reaction assay using MGB probe technology.
A real-time PCR assay was developed to quantify human herpesvirus-7 (HHV-7) genome based on TaqMan technology using the new MGB probe. Primers and probe were chosen in the conserved U100 gene. Plasmid containing the sequence of interest was constructed for the standardisation of the method and to assess its sensitivity. This HHV-7 genomic quantitation assay has a threshold sensitivity of fourteen equivalent genome copy number (EqCop) per reaction. This method was applied to the quantitation of HHV-7 in the peripheral blood mononuclear cells (PBMCs) obtained from 31 healthy subjects. Eighty seven per cent had HHV-7 positive detection in the PBMCs with a viral load ranging from 275 to 14545 EqCop per million of cells. This method presents interesting characteristics with a wide range of quantitation, a good sensitivity, and constitutes a new tool for the study of HHV-7 infection in vivo and in vitro.
D000328:Adult; D004279:DNA, Viral / Q000097:blood*; D005260:Female; D016199:Herpesvirus 7, Human / Q000302:isolation & purification*; D006801:Humans; D008297:Male; D015345:Oligonucleotide Probes / Q000235:genetics*; D016133:Polymerase Chain Reaction / Q000379:methods* / Q000592:standards; D019349:Roseolovirus Infections / Q000821:virology*; D012680:Sensitivity and Specificity; D019914:Taq Polymerase; D019562:Viral Load
D004279:DNA, Viral; D015345:Oligonucleotide Probes; D019914:Taq Polymerase