title: Comparison of the sensitivity and specificity of real-time PCR and in situ hybridization in HPV16 and 18 detection in archival cervical cancer specimens.
abstract: The aim of this study was to analyze the correlation between real-time PCR (RT-PCR) treated as a reference method and in situ hybridization with tyramide amplification system (ISH-TSA) in the detection of HPV16 and 18 infection and the assessment of viral genome status. The study was performed on cervical cancer biopsies fixed in 10% neutral buffered formalin and embedded in paraffin obtained from 85 women. TaqMan-based 5'exonuclease RT-PCR with type-specific primers was used to assess HPV16 and 18 infections and genome status. Viral infection and genome status was also assessed by ISH-TSA. RT-PCR revealed 76 (89.4%), and ISH-TSA 81 (95.3%) cancers with HPV16 and 18 infections. The ISH-TSA sensitivity and specificity were: 96.1% and 11.1% compared to RT-PCR. The difference between these techniques in HPV detection was significant (p = 0.000). Among 76 HPV16/18 positive cancers in RT-PCR, there were 30 (39.5%) with integrated and 46 (60.5%) with mixed viral genome form. According to ISH-TSA, there were 39 (51.3%) samples with integrated and 37 with mixed form (48.7%). The sensitivity and specificity of ISH-TSA in genome status assessment were 70.0% and 60.9%, respectively. The difference between RT-PCR and ISH-TSA in genome state detection was not statistically significant (p = 0.391). These results suggest that ISH-TSA shows insufficient specificity in HPV detection for use in clinical practice. However, this assay could be applied for viral genome status assessment.
authors: Department of Applied Radiobiology, Center of Oncology, Krakow Division, Krakow, Poland. z5biesag@cyfronet.pl
authors: Beata
authors: B
authors: Biesaga
authors: Sława
authors: S
authors: Szostek
authors: Małgorzata
authors: M
authors: Klimek
authors: Jerzy
authors: J
authors: Jakubowicz
authors: Joanna
authors: J
authors: Wysocka
mesh_terms: D018070:Biological Specimen Banks; D005260:Female; D018628:Gene Dosage / Q000235:genetics; D016679:Genome, Viral / Q000235:genetics; D052162:Human papillomavirus 16 / Q000235:genetics* / Q000302:isolation & purification; D052161:Human papillomavirus 18 / Q000235:genetics* / Q000302:isolation & purification; D006801:Humans; D017403:In Situ Hybridization / Q000379:methods*; D008875:Middle Aged; D030361:Papillomavirus Infections / Q000235:genetics / Q000473:pathology / Q000821:virology; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D012015:Reference Standards; D012680:Sensitivity and Specificity; D002583:Uterine Cervical Neoplasms / Q000473:pathology / Q000821:virology*