„Nested“ PCR for Human Papillomavirus Detectionand Typing in Samples from Cervical Lesionsof Gynecological Patients

Overview

Objective:
The authors tested PCR – so called nested (N-PCR) for the detection of HPV in cervicallesions of uterus. This method is based on repeated amplifi cation of specifi c viral DNA fragments.The sensitivity of N-PCR was compared to the sensitivity of the hybridization method. In the secondpart of the experiment positive samples underwent HPV typing by PCR.Background: The effectiveness of N-PCR for HPV detection was verifi ed on 30 samples fromcytologically/histologically suspected cervical lesions. In these cases, previous detection byhybridization method was unsuccessful. The second group consisted of 21 samples acquired byconisation, in which HPV presence was confi rmed by hybridization method. Post surgery HPVdetection using hybridization technique was negative in this group of patients.Results: By means of N-PCR the presence of HPV DNA was confi rmed in all 30 samples fromthe fi rst group (100%) and in 8 (38%) cases from the group of samples obtained after conisation.Type-specifi c PCR detection indicated HPV type 16 in 20 cases, HPV type 18 in 6 cases (from thefi rst series). In the remaining 4 cases typing for HPV 16, 18, 31 and 33 was negative. In the secondseries all 8 samples were HR-HPV type 16.Conclusion: At the present time, N-PCR is probably the most sensitive HPV detection method. Theonly real disadvantages of the given technique are relatively high costs of diagnostics equipmentand the requirement of highly qualifi ed personel.

Key words:
nested PCR, HPV detection, typing