„Nested“ PCR for Human Papillomavirus Detection
and Typing in Samples from Cervical Lesions
of Gynecological Patients
Kurillová Ľ.1, Šimko J.2, Holomáň K.2, Šuška P.2, Rusnák I.3
1Laboratórium molekulárnej biológie, Bratislava, odborný garant MUDr. RNDr. J. Šimko, PhD. 2II. gynekologicko-pôrodnícka klinika FN, Bratislava, prednosta prof. MUDr. K. Holomáň, CSc3Katedra gynekológie a pôrodníctva SZU, Bratislava, prednosta prof. MUDr. J. Štencl, CSc. |
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Summary:
Objective: The authors tested PCR – so called nested (N-PCR) for the detection of HPV in cervical
lesions 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 second
part 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 from
cytologically/histologically suspected cervical lesions. In these cases, previous detection by
hybridization method was unsuccessful. The second group consisted of 21 samples acquired by
conisation, in which HPV presence was confi rmed by hybridization method. Post surgery HPV
detection 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 from
the 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 the
fi rst series). In the remaining 4 cases typing for HPV 16, 18, 31 and 33 was negative. In the second
series all 8 samples were HR-HPV type 16.
Conclusion: At the present time, N-PCR is probably the most sensitive HPV detection method. The
only real disadvantages of the given technique are relatively high costs of diagnostics equipment
and the requirement of highly qualifi ed personel.
Key words:
nested PCR, HPV detection, typing
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