Hybrid capture II and polymerase chain reaction for identifying HPV infections in samples collected in a new collection medium: a comparison.
OBJECTIVE
To compare the performance of human papillomavirus (HPV) DNA detection by polymerase chain reaction (PCR) and Hybrid Capture II (HCII) test (Digene, Gaithersburg, Maryland, U.S.A.) in residual cells left in the collection vials of the DNACitoliq system (Digene Brasil, São Paulo, Brazil).


METHODS
A series of 263 cervical samples collected for liquid-based cytology with the DNACitoliq system was tested for oncogenic HPV types first with HCII and subsequently with PCR. After DNA purification with GFX Genomic Blood DNA Purification Kit (Amersham, Piscataway, New Jersey, U.S.A.), PCR was performed using AmpliTaq Gold DNA polymerase (Applied Biosystems). PGMY09/11 L1 consensus primers and GH20/PCO4 primers for human beta-globin target were coamplified.


RESULTS
Altogether, 260 samples were positive for beta-globin, and 3 negative ones were excluded from the analysis. PCR and HCII yielded concordant results in 199 cases (76.5%) (102 positive and 97 negative), with Cohen's kappa of .577 (95% CI .477-.677) and weighted kappa of .733 (95% CI .659-.791). HPV prevalence in different categories of cytologic abnormalities was practically identical with HCII and PCR assays (P=.989). Among the 61 (23.5%) discrepant cases, 28 samples were HCII+/PCR- cases. Of these, 27 of 28 samples showed a low viral load, and 1 had an intermediate viral load.


CONCLUSIONS
The data suggest that residual material from the DNACitoliq system adequately preserves HPV DNA for detection by HCII and PCR, with performance similar to that of specimen transport medium.
Pathology Division, Adolfo Lutz Institute, São Paulo, Brazil.
Suely
S
Nonogaki
Alda
A
Wakamatsu
Adhemar
A
Longatto Filho
Sonia Maria Miranda
SM
Pereira
Maria Lúcia
ML
Utagawa
Venâncio Avancini
VA
Ferreira Alves
Celso
C
Di Loreto
Marina Yoshiê
MY
Sakamoto Maeda
Temístocles P
TP
Lima
Cecília
C
Roteli-Martins
Kari
K
Syrjänen
D000293:Adolescent; D000328:Adult; D001706:Biopsy / Q000379:methods; D015344:DNA Probes, HPV; D004279:DNA, Viral / Q000032:analysis; D005260:Female; D005838:Genotype; D006801:Humans; D008875:Middle Aged; D027383:Papillomaviridae / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D030361:Papillomavirus Infections / Q000175:diagnosis* / Q000821:virology; D016133:Polymerase Chain Reaction; D011933:Reagent Kits, Diagnostic; D015203:Reproducibility of Results; D013048:Specimen Handling / Q000295:instrumentation / Q000379:methods*
D015344:DNA Probes, HPV; D004279:DNA, Viral; D011933:Reagent Kits, Diagnostic