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Updated: Jan 22, 2026

Multiplex PCR and Reverse Line Blot Hybridization Assay mPCR/RLB
Published on: August 6, 2011
Identification by high-throughput sequencing of HPV variants and quasispecies that are untypeable by linear reverse
Lucie Molet1, Delphine Girlich2, Rémy A Bonnin2
1Laboratoire de Virologie, Assistance Publique-Hôpitaux de Paris (AP-HP), Hôpitaux Universitaires Paris Sud, Hôpital Paul Brousse, Villejuif, France; INSERM UMR-996 « Inflammation, Chimiokines et Immunopathologies », Université Paris Sud, Université Paris Saclay, LabEx Lermit, Faculté de Médecine, Clamart, France.
Abstract:
The linear reverse blotting assays are valid methods for accurate human papillomavirus (HPV) typing required to manage women at risk of developing cervical cancer. However, some samples showed a positive signal in HPV lines but failed to display a positive signal in subsequent typing lines (designated as HPV-X), which indicate that certain types were not available on the respective typing blots. The aim of this study is to elucidate the types or variants of HPV through the high-throughput sequencing (HTS) of 54 ASCUS cervical samples in which the viruses remained untypeable with INNO LiPA HPV® assays. Low-risk (LR)-HPV types (HPV6, 30, 42, 62, 67, 72, 74, 81, 83, 84, 87, 89, 90 and 114), high-risk (HR)-HPV35 and possibly (p)HR-HPV73 were detected among HPV-X. Individual multiple infections (two to seven types) were detected in 40.7% of samples. Twenty-two specimens contained variants characterised by 2-10 changes. HPV30 reached the maximal number of 17 variants with relative abundance inferior or equal to 2.7%. The presence of L1 quasispecies explains why linear reverse blotting assays fail when variants compete or do not match the specific probes. Further studies are needed to measure the LR-HPV quasispecies dynamics and its role during persistent infection.
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