Increase in viral load, viral integration, and gain of telomerase genes during uterine cervical carcinogenesis can be simultaneously assessed by the HPV 16/18 MLPA-assay.
Oncogenic human papillomavirus (HPV) infection is the most important risk factor in cervical carcinogenesis cases; high viral loads, viral integration into the host genome, and gain of the telomerase-related genes, TERT and TERC, are all factors associated with progression to cancer. A recently developed multiparameter HPV 16/18 multiplex ligation-dependent probe amplification (MLPA) assay, which allows the simultaneous assessment of these factors, was applied to a series of 67 normal and (pre)malignant frozen uterine cervical samples, as well as to 91 cytological preparations, to test the ability of the MLPA assay to identify high-risk lesions on the basis of these factors. Validation was performed using quantitative PCR, the PapilloCheck and fluorescence in situ hybridization. Only 5 out of 37 normal tissue samples or low-grade cervical lesions (ie, CIN1 and condyloma) showed either an HPV16 viral load higher than 25 copies per cell, viral integration, and/or gain of one of the telomerase-related genes, whereas for the high-grade cervical lesions, one or more of these risk factors was found in 25 of 30 cases. The HPV MLPA assay showed a sensitivity of 83% and a specificity of 86% in frozen cervical specimens. Furthermore, the feasibility of the MLPA assay was shown for cytological samples, where in 57% of high-grade squamous intraepithelial lesion cases, the high-risk factors were detected using this assay.
Theelen
Wendy
W
Departments of Molecular Cell Biology, GROW - School for Oncology and Developmental Biology, Maastricht University Medical Center, Maastricht, The Netherlands.
Speel
Ernst-Jan M
EJ
Herfs
Michael
M
Reijans
Martin
M
Simons
Guus
G
Meulemans
Els V
EV
Baldewijns
Marcella M
MM
Ramaekers
Frans C S
FC
Somja
Joan
J
Delvenne
Philippe
P
Hopman
Anton H N
AH
D000328:Adult; D000368:Aged; D002294:Carcinoma, Squamous Cell / Q000175:diagnosis / Q000209:etiology; D002471:Cell Transformation, Neoplastic / Q000235:genetics / Q000473:pathology; D004279:DNA, Viral / Q000235:genetics; D005240:Feasibility Studies; D005260:Female; D052162:Human papillomavirus 16 / Q000302:isolation & purification*; D052161:Human papillomavirus 18 / Q000302:isolation & purification*; D006801:Humans; D017404:In Situ Hybridization, Fluorescence; D008875:Middle Aged; D021141:Nucleic Acid Amplification Techniques; D030361:Papillomavirus Infections / Q000175:diagnosis* / Q000209:etiology; D016133:Polymerase Chain Reaction; D019098:Telomerase / Q000235:genetics* / Q000378:metabolism; D002583:Uterine Cervical Neoplasms / Q000175:diagnosis* / Q000209:etiology; D014599:Uterus / Q000378:metabolism / Q000473:pathology; D019562:Viral Load; D016662:Virus Integration; D055815:Young Adult; D002578:Uterine Cervical Dysplasia / Q000175:diagnosis / Q000209:etiology
D004279:DNA, Viral; C509186:TERT protein, human; D019098:Telomerase
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