title: Comparison of SPF10 real-time PCR and conventional PCR in combination with the INNO-LiPA HPV Genotyping Extra assay for the detection and typing of human papillomavirus in cervical samples.
abstract: The novel SPF10 real-time PCR assay allows the simultaneous amplification and detection of the HPV target. That way, LiPA analysis of the HPV-negative samples can be avoided, reducing workload and cost. This study aims to evaluate the performance of the SPF10 real-time PCR in combination with the LiPA assay for HPV detection and typing in cervical samples. Thirty-nine cervical samples were subjected to the SPF10 conventional PCR in combination with the LiPA assay. Subsequently, the SPF10 real-time PCR was performed to enable the comparison between the SPF10 conventional and the real-time PCR results. In case of discrepancy, the samples were subjected to the CLART HPV2 assay. As a result, 27 out of 39 samples were identified as HPV-positive by the SPF10 real-time PCR and were genotyped further by the LiPA assay. Twenty samples (74.1%) showed an absolute agreement between the conventional and real-time SPF10 PCR (concordant), three (11.1%) displayed additional or fewer types (compatible), two (7.4%) did not show any similarity between both assays (discordant) and the remaining two (7.4%) were LiPA-negative. The two assays showed an excellent strength of agreement for individual (κ=0.932) and multiple genotype detection (κ=0.834). In conclusion, the two SPF10 PCR methods are comparable. Therefore, the SPF10 real-time PCR with subsequent LiPA could be used for the detection and genotyping of HPV in cervical samples.
authors: Applied Molecular Biology Research (AMBIOR) Group, Laboratory of Cell Biology and Histology, University of Antwerp, Groenenborgerlaan 171, B2020 Antwerp, Belgium; Vaccine & Infectious Diseases Institute (VAXINFECTIO), University of Antwerp, Universiteitsplein 1, B2610 Antwerp, Belgium. Electronic address: Isabel.Micalessi@ua.ac.be.
authors: M I
authors: MI
authors: Micalessi
authors: G A
authors: GA
authors: Boulet
authors: S
authors: S
authors: Pillet
authors: J
authors: J
authors: Jacquet
authors: B
authors: B
authors: Pozzetto
authors: J J
authors: JJ
authors: Bogers
authors: T
authors: T
authors: Bourlet
mesh_terms: D002584:Cervix Uteri / Q000821:virology*; D005260:Female; D006801:Humans; D025202:Molecular Diagnostic Techniques / Q000379:methods*; D027383:Papillomaviridae / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification*; D030361:Papillomavirus Infections / Q000175:diagnosis* / Q000821:virology*; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*
keywords: Cervical specimen; Genotype; Human papillomavirus; LiPA; Real-time PCR; SPF10