Detection of human papilloma virus (HPV) genomes by the primed in situ (PRINS) labelling technique.
Primed in situ Labelling, a technique based on primer mediated DNA synthesis, has become a useful tool in cytogenetics, especially for chromosome mapping, banding and the investigation of sequence organization in fresh metaphase preparations. Its application in the routine surgical pathology laboratory has been hampered by the fact that the technique did not work on paraffin-embedded, formalin-fixed tissue. We investigated cervical biopsies (n = 20) with morphological signs of HPV infection and found that the PRINS method is at least as sensitive as a classical in situ hybridization assay for detecting HPV DNA in paraffin-embedded, formalin-fixed tissue. In all investigated cases (n = 20), HPV DNA was found by both methods. The PRINS method was able to demonstrate HPV DNA not only in superficial koilocytotic squamous cells but also in non-koilocytotic cells in the deeper spinous cell layers, and even in some basal cells. We describe an economical protocol using conventional consensus HPV oligonucleotide DNA primers. The described method is rapid (approximately 3 hours) and easy to perform for screening and subtyping HPV infection in the routine surgical pathology laboratory.
Ramael
M
M
Royal Antwerp Micrographical Society, Belgium. marc.ramael@skynet.be
Van Steelandt
H
H
Stuyven
G
G
Van Steenkiste
M
M
Degroote
J
J
D017931:DNA Primers / Q000737:chemistry; D004279:DNA, Viral / Q000032:analysis; D005260:Female; D016679:Genome, Viral; D006801:Humans; D027383:Papillomaviridae / Q000235:genetics* / Q000302:isolation & purification; D030361:Papillomavirus Infections / Q000175:diagnosis / Q000821:virology*; D020401:Primed In Situ Labeling / Q000379:methods*; D012680:Sensitivity and Specificity; D014412:Tumor Virus Infections / Q000175:diagnosis / Q000821:virology*; D002577:Uterine Cervical Diseases / Q000175:diagnosis / Q000821:virology*
D017931:DNA Primers; D004279:DNA, Viral