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

• Published in: Journal of virological methods Vol. 194; no. 1-2; pp. 113 - 117
• Main Authors: Micalessi, M.I., Boulet, G.A., Pillet, S., Jacquet, J., Pozzetto, B., Bogers, J.J., Bourlet, T.
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier B.V 01.12.2013
• Subjects: Cervical specimen; Cervix Uteri - virology; Female; Genotype; genotyping; Human papillomavirus; Humans; LiPA; Molecular Diagnostic Techniques - methods; Papillomaviridae; Papillomaviridae - classification; Papillomaviridae - genetics; Papillomaviridae - isolation & purification; Papillomavirus Infections - diagnosis; Papillomavirus Infections - virology; quantitative polymerase chain reaction; Real-time PCR; Real-Time Polymerase Chain Reaction - methods; SPF10; Genotype; Human papillomavirus; LiPA; Real-time PCR; Cervical specimen; SPF10
• ISSN: 0166-0934; 1879-0984
• DOI: 10.1016/j.jviromet.2013.08.013

•Simultaneous amplification and detection of HPV target by a SPF10 real-time PCR.•Reduction of workload and cost by avoiding LiPA step for HPV-negative samples.•Assessment of the performance of the SPF10 real-time PCR using cervical samples.•High comparability between SPF10 real-time PCR and conventional PCR. 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.