Human papillomavirus detection and typing using a nested-PCR-RFLP assay.
BACKGROUND
It is clinically important to detect and type human papillomavirus (HPV) in a sensitive and specific manner.


OBJECTIVE
Development of a nested-polymerase chain reaction-restriction fragment length polymorphism (nested-PCR-RFLP) assay to detect and type HPV based on the analysis of L1 gene.


METHODS
Analysis of published DNA sequence of mucosal HPV types to select sequences of new primers. Design of an original nested-PCR assay using the new primers pair selected and classical MY09/11 primers. HPV detection and typing in cervical samples using the nested-PCR-RFLP assay.


RESULTS
The nested-PCR-RFLP assay detected and typed HPV in cervical samples. Of the total of 128 clinical samples submitted to simple PCR and nested-PCR for detection of HPV, 37 (28.9%) were positive for the virus by both methods and 25 samples were positive only by nested-PCR (67.5% increase in detection rate compared with single PCR). All HPV positive samples were effectively typed by RFLP assay.


CONCLUSIONS
The method of nested-PCR proved to be an effective diagnostic tool for HPV detection and typing.
Postgraduate Program in Genetics and Molecular Diagnosis, Universidade Luterana do Brasil, Canoas, RS, Brazil.
Janaina
J
Coser
Thaís da Rocha
Tda R
Boeira
André Salvador Kazantzi
AS
Fonseca
Nilo
N
Ikuta
Vagner Ricardo
VR
Lunge
D001483:Base Sequence; D002584:Cervix Uteri / Q000821:virology*; D017931:DNA Primers / Q000032:analysis; D004279:DNA, Viral / Q000032:analysis; D005260:Female; D006801:Humans; D027383:Papillomaviridae / Q000145:classification / Q000235:genetics*; D030361:Papillomavirus Infections / Q000175:diagnosis* / Q000821:virology; D016133:Polymerase Chain Reaction; D012150:Polymorphism, Restriction Fragment Length; D012680:Sensitivity and Specificity
D017931:DNA Primers; D004279:DNA, Viral