title: Laser micro-dissection and qPCR for identifying specific HPV types responsible for malignancy in penile lesions.
abstract: The aim of the study was to identify specific human papillomavirus (HPV) type responsible for malignancy in penile tissue samples using laser micro-dissection and TaqMan quantitative real-time PCR (qPCR). The study was based on two pre-malignant and seven malignant penile tissue samples and laser micro-dissection was performed on all. Genotyping was performed on whole tissue sections and laser micro-dissection samples using qPCR. Two whole tissue section samples were HPV negative while seven were HPV positive. In four samples that were single HPV infections with whole tissue section PCR, identical HPV types were confirmed with laser micro-dissection PCR. Clearly confirming that the single HPV type detected is responsible for malignancy. In two samples that had multiple HPV infections with whole tissue section PCR, only one HPV type with the highest viral load was detected with laser micro-dissection PCR, suggesting that the HPV type with the highest viral load is most likely the cause of that particular lesion. HPV 11 and/or HPV 16 were the only types detected with laser micro-dissection PCR in these cases, compared to multiple HPV types (HPV 11, HPV 16, HPV 18, HPV 31, HPV 33, HPV 35, and HPV 39) initially detected with whole tissue section PCR. HPV 11 was associated with verrucous lesions while HPV 16 was associated with squamous cell carcinoma and PIN 3 lesions. This study confirms that laser micro-dissection and qPCR are essential tools in identifying the HPV types responsible for malignancy in penile lesions, particularly in samples with multiple infections.
authors.affiliation: Department of Virology, Sefako Makgatho Health Sciences University/National Health Laboratory Service, Medunsa, Pretoria, South Africa.
authors.forename: Ramokone L
authors.initials: RL
authors.lastname: Lebelo
authors.affiliation: Laboratory of Cell Biology and Histology, University of Antwerp, Antwerp, Belgium.
authors.forename: Sofie
authors.initials: S
authors.lastname: Thys
authors.affiliation: Department of Molecular Diagnostics, Laboratory for Clinical Pathology AML, Sonic HealthCare Benelux, Antwerp, Belgium.
authors.forename: Ina
authors.initials: I
authors.lastname: Benoy
authors.affiliation: Department of Molecular Diagnostics, Laboratory for Clinical Pathology AML, Sonic HealthCare Benelux, Antwerp, Belgium.
authors.forename: Christophe E
authors.initials: CE
authors.lastname: Depuydt
authors.affiliation: Laboratory of Cell Biology and Histology, University of Antwerp, Antwerp, Belgium.|Department of Molecular Diagnostics, Laboratory for Clinical Pathology AML, Sonic HealthCare Benelux, Antwerp, Belgium.
authors.forename: John-Paul
authors.initials: JP
authors.lastname: Bogers
authors.affiliation: Division of Anatomical Pathology, Sefako Makgatho Health Sciences University/National Health Laboratory Service, Medunsa, Pretoria, South Africa.
authors.forename: Meshack N
authors.initials: MN
authors.lastname: Bida
authors.affiliation: Department of Virology, Sefako Makgatho Health Sciences University/National Health Laboratory Service, Medunsa, Pretoria, South Africa.
authors.forename: M Jeffrey
authors.initials: MJ
authors.lastname: Mphahlele
mesh_terms: D002294:Carcinoma, Squamous Cell / Q000175:diagnosis / Q000473:pathology / Q000821:virology; D018289:Carcinoma, Verrucous / Q000175:diagnosis / Q000473:pathology / Q000821:virology; D004279:DNA, Viral / Q000032:analysis; D005838:Genotype; D052140:Human papillomavirus 11 / Q000145:classification / Q000235:genetics* / Q000302:isolation & purification; D052162:Human papillomavirus 16 / Q000145:classification / Q000235:genetics* / Q000302:isolation & purification; D006801:Humans; D061346:Laser Capture Microdissection; D008297:Male; D008875:Middle Aged; D030361:Papillomavirus Infections / Q000175:diagnosis / Q000821:virology*; D016612:Paraffin Embedding; D010412:Penile Neoplasms / Q000473:pathology / Q000821:virology*; D060888:Real-Time Polymerase Chain Reaction; D019562:Viral Load
chemical_list: D004279:DNA, Viral
keywords: human papillomavirus; laser micro-dissection; real time PCR; squamous cell carcinoma; verrucous carcinoma