PAPILLOMAVIRUS SCREENING IN CERVICAL CELL SAMPLES USING DUAL-LABEL DOT-BLOT ANALYSIS

The presence of human papillomavirus (HPV) in cervical cells is closel y related to the development of cervical carcinoma. Detection of virus may be by Southern blot, dot blot or the highly sensitive polymerase chain reaction. Whatever method is employed, there are problems of fal se negatives due to poor clinical samples in which the DNA may be degr aded or is absent altogether. Here we describe a new method of dual la belling for dot blots using a P-32-labelled probe for HPV and a S-35-l abelled probe for human actin genes. The samples were counted on a Bet aplate(TM) flat-bed scintillation counter and the data analysed to sep arate the activities of the two isotopes. The counts from the actin pr obe show whether human DNA is present or not and false negatives from this cause may thereby be eliminated. The counts due to HPV when compa red with those for actin give a quantitative measure of HPV abundance for the particular sample and this may have clinical relevance.