Direct in situ PCR allows rapid and sensitive detection of high risk human papillomavirus in cytologic specimens and formalin-fixed paraffin tissues by fluorescent labelling.
We developed a rapid, sensitive and robust high risk human papillomavirus (HR HPV) detection protocol based on direct in situ PCR technology and fluorochrome-modified nucleotides on cytologic specimens (cell smears) and on HPV infected tissues (CIN III). Reproducible results on both cytologic specimens and paraffin-embedded tissues were obtained, providing a powerful tool for clinical investigation on HR HPV infection. Quantitative PCR performed on the same tissue sections adjacent to those used for in situ techniques allowed us to establish the sensitivity of our methods, able to detect rare copies (about 15 in our paraffin-embedded tissues) of HPV.
Comar
M
M
Istituto di Igiene e Medicina Preventiva, Universta di Trieste, Trieste, Italy.
Spano
A
A
Canova
S
S
Bogoni
S
S
Marziliano
N
N
Cernigoi
E
E
Boniotto
M
M
Amoroso
A
A
Parodi
S
S
Campello
C
C
Crovella
S
S
D005260:Female; D005557:Formaldehyde; D006367:HeLa Cells; D006801:Humans; D017404:In Situ Hybridization, Fluorescence / Q000379:methods; D027383:Papillomaviridae / Q000235:genetics / Q000302:isolation & purification*; D016612:Paraffin Embedding; D016133:Polymerase Chain Reaction / Q000379:methods*; D012307:Risk Factors; D012680:Sensitivity and Specificity; D002578:Uterine Cervical Dysplasia / Q000821:virology*
D005557:Formaldehyde