["Nested" PCR for human papillomavirus detection and typing in samples from cervical lesions of gynecological patients].
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 amplification of specific 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 verified 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 confirmed 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 confirmed in all 30 samples from the first group (100%) and in 8 (38%) cases from the group of samples obtained after conisation. Type-specific PCR detection indicated HPV type 16 in 20 cases, HPV type 18 in 6 cases (from the first 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.


CONCLUSIONS
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 qualified personel.
Kurillová
L
L
Laboratórium molekulárnej biológie, Bratislava.
Simko
J
J
Holomán
K
K
Suska
P
P
Rusnák
I
I
D002584:Cervix Uteri / Q000821:virology*; D004279:DNA, Viral / Q000032:analysis; D005260:Female; D006801:Humans; D027383:Papillomaviridae / Q000145:classification / Q000302:isolation & purification*; D016133:Polymerase Chain Reaction; D012680:Sensitivity and Specificity; D002583:Uterine Cervical Neoplasms / Q000821:virology*; D002578:Uterine Cervical Dysplasia / Q000821:virology*
D004279:DNA, Viral
"Nested" PCR a jej vyuzitie pri detekcii a typizácii papilomavírusovej infekcie vo vzorkách z cervikálnych lézií gynekologických pacientok.