New approach to human high-risk papillomavirus (HR-HPV) genotyping.
Human high-risk papillomaviruses (HR-HPVs) are involved in the induction of invasive cervical cancer. The aim of this study was to introduce a simple, semi-automated and reproducible approach suitable for HR-HPV detection in clinical practice. The procedure is based on DNA isolation, nested polymerase chain reaction, single strand conformational polymorphism and evaluation of HR-HPV genotypes with Gel-Pro software. The clinical performance of the new approach was assessed in two different patient materials: 1) cervical smears with cytological classification Pap2-3 or Pap3 lacking nuclear atypia (anisonucleosis and polychromasia) or koilocytotic atypia and without any previous therapy 2) formalin-fixed, paraffin-embedded cervical carcinoma and lymph node sections. Using the new approach we detected HR-HPV DNA in 64% patient samples cytologically classified as Pap2-3 or Pap3 respectively and in 80% formalin-fixed, paraffin-embedded lymph node sections histologically classified as lymph nodes without carcinoma cell infiltration. The combination of methods described in this study results in increased sensitivity of HR-HPV identification allowing detection of HPV DNA in a very small amount of target DNA so that it can be widely used in distinguishing the pre- malignant lesions and in determination of invading carcinoma cells to lymph nodes in patients with advanced cervical cancer. The new approach is useful in unambiguous HR-HPV genotyping even in double-HPV infection.
Weismanová
E
E
Department of Clinical Genetics, St. Elizabeth Cancer Institute, 812 50 Bratislava 1, Slovak Republic. eweisman@ousa.sk
Weismann
P
P
Vizváryová
M
M
Lehotská
V
V
Krizanová
O
O
Repiská
V
V
Kausitz
J
J
D001331:Automation; D002277:Carcinoma / Q000175:diagnosis* / Q000473:pathology; D002584:Cervix Uteri / Q000821:virology; D004279:DNA, Viral / Q000032:analysis*; D005260:Female; D005838:Genotype; D006801:Humans; D008207:Lymphatic Metastasis; D027383:Papillomaviridae / Q000235:genetics / Q000302:isolation & purification*; D030361:Papillomavirus Infections / Q000175:diagnosis* / Q000473:pathology; D016133:Polymerase Chain Reaction / Q000379:methods*; D018807:Polymorphism, Single-Stranded Conformational; D011230:Precancerous Conditions / Q000175:diagnosis; D012680:Sensitivity and Specificity; D012984:Software; D014412:Tumor Virus Infections / Q000175:diagnosis* / Q000473:pathology; D002583:Uterine Cervical Neoplasms / Q000175:diagnosis / Q000473:pathology
D004279:DNA, Viral