Novel method for detection, typing, and quantification of human papillomaviruses in clinical samples.
We report the development of a novel detection and typing methodology for human papillomaviruses (HPV) based on real-time PCR with the self-probing fluorescent primers known as Scorpions. This technique is quick, simple, specific, sensitive, and capable of estimating viral load per cell. We report the results of over 100 typing reactions performed on cell lines, biopsies, and cervical cytobrush samples which, when compared to the current reference HPV detection and typing technique, present a kappa value of 0.89. We further report preliminary data suggesting a relationship between viral load per cell and grade of cervical disease.
Hart
K W
KW
Section of Infection and Immunity, Department of Medicine, University of Wales College of Medicine, Heath Park, Cardiff CF14 4XX, United Kingdom. HartKW@cf.ac.uk
Williams
O M
OM
Thelwell
N
N
Fiander
A N
AN
Brown
T
T
Borysiewicz
L K
LK
Gelder
C M
CM
D002584:Cervix Uteri / Q000821:virology; D017931:DNA Primers; D004279:DNA, Viral / Q000032:analysis; D005260:Female; D006801:Humans; D007124:Immunoenzyme Techniques; D027383:Papillomaviridae / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification* / Q000502:physiology; D030361:Papillomavirus Infections / Q000821:virology*; D016133:Polymerase Chain Reaction / Q000379:methods*; D014407:Tumor Cells, Cultured; D014412:Tumor Virus Infections / Q000821:virology*; D019562:Viral Load
D017931:DNA Primers; D004279:DNA, Viral