Molecular strategies for identifying human papillomavirus infection in routinely processed samples: focus on paraffin sections.
OBJECTIVE
To optimize the protocols of human papillomavirus (HPV) detection in clinical samples, we used polymerase chain reaction (PCR)-based techniques in paraffin-embedded tissue sections and compared the results with those obtained with PCR and Hybrid Capture II (HC2) performed in liquid-based cytology (LBC) preservation medium.


METHODS
Forty-five consecutive cervical biopsy specimens were taken from women participating in the ongoing Latin American Screening Study at Leonor Mendes de Barros Hospital, São Paulo, Brazil, during 2003 and 2004. The biopsy specimens were analyzed for HPV by a modified GP5+/GP6+ PCR protocol, and the results were compared with those obtained by PGMY PCR and HC2 in samples collected in LBC preservation medium.


RESULTS
beta-Globin was detected in 100% of the multiplex PCR system from LBC samples and 66.7% with PCO4+/PCO3+ PCR in biopsy specimens. Of the three methods, PGMY PCR system and HC2 were equally effective in detecting HPV; both detected 13 cases in 45 samples (28.9%). The GP5+/GP6+ PCR applied in biopsy specimens showed a 20% HPV detection rate (9/45).


CONCLUSIONS
Our PCR protocols worked reasonably well and allowed us to compare the three molecular methods with histological and cytological findings. The reproducibility of the results makes the technique applicable in archival materials.
Nonogaki
Suely
S
Pathology Division of Adolfo Lutz Institute, São Paulo, Brazil.
Wakamatsu
Alda
A
Filho
Adhemar Longatto
AL
Roteli-Martins
Cecília
C
di Loreto
Celso
C
Maeda
Marina Yoshiê Sakamoto
MY
Utagawa
Maria Lúcia
ML
Pereira
Sônia Maria Miranda
SM
Polli
Sabrina
S
Alves
Venâncio A F
VA
Syrjänen
Kari
K
D000293:Adolescent; D000328:Adult; D001706:Biopsy; D001938:Brazil; D002584:Cervix Uteri / Q000473:pathology / Q000821:virology; D003955:Diagnostic Tests, Routine; D005260:Female; D005914:Globins / Q000032:analysis; D006801:Humans; D008875:Middle Aged; D025202:Molecular Diagnostic Techniques; D027383:Papillomaviridae / Q000302:isolation & purification*; D030361:Papillomavirus Infections / Q000175:diagnosis* / Q000473:pathology / Q000821:virology*; D010232:Paraffin; D016133:Polymerase Chain Reaction; D012597:Sclerosing Solutions; D019562:Viral Load
D012597:Sclerosing Solutions; D010232:Paraffin; D005914:Globins