High prevalence of HPV 51 in an unvaccinated population and implications for HPV vaccines

Human papillomavirus (HPV) is detected in 99.7% of cervical cancers. Current vaccines target types 16 and 18. Prior to vaccination implementation, a prospective cohort study was conducted to determine baseline HPV prevalence in unvaccinated women in Wales; after HPV16 and HPV18, HPV 51 was found to be most prevalent. This study aimed to re-assess the unexpected high prevalence of HPV 51 and consider its potential for type-replacement. Two hundred HPV 51 positive samples underwent re-analysis by repeating the original methodology using HPV 51 GP5+/6+ PCR-enzyme immunoassay, and additionally a novel assay of HPV 51 E7 PCR. Data were correlated with age, social deprivation and cytology. Direct repeat of HPV 51 PCR-EIA identified 146/195 (75.0%) samples as HPV 51 positive; E7 PCR identified 166/195 (85.1%) samples as HPV 51 positive. HPV 51 prevalence increased with cytological grade. The prevalence of HPV 51 in the pre-vaccinated population was truly high. E7 DNA assays may offer increased specificity for HPV genotyping. Cross-protection of current vaccines against less-prevalent HPV types warrants further study. This study highlights the need for longitudinal investigation into the prevalence of non-vaccine HPV types, especially those phylogenetically different to vaccine types for potential type-replacement. Ongoing surveillance will inform future vaccines

Human papillomavirus negative but dyskaryotic cervical cytology: re-analysis of molecular testing

Background Evaluation of molecular Human Papillomavirus (HPV) testing into UK Cervical Screening Programmes is underway. In South Wales the current HPV prevalence in women attending routine screening is 13.5% with 76.3% HR HPV positive in cases with reported dyskaryotic cervical cytology [Hibbitts S, Jones J, Powell N, Dallimore N, McRea J, Beer H, et al. Human papillomavirus prevalence in women attending routine cervical screening in South Wales, UK: a cross-sectional study. Br J Cancer 2008;99(December (11)):1929–33]. Objectives The aim of this study was to re-analyse the 23.7% cases with reported dyskaryotic cytology that were HR HPV negative (n = 52 out of 219 in a population of 10,000). Study design Three procedures were performed: (i) GP5+/GP6+ PCR-EIA repeat on original DNA extracts; (ii) DNA extraction and GP5+/GP6+ HPV PCR-EIA; (iii) DNA extraction and HPV typing using Greiner Bio-One™ PapilloCheck DNA microarray. Results 51 out of 52 samples were re-analysed. Direct repeat HPV PCR-EIA identified 24% (n = 12/51) of samples positive for HR HPV. Re-extracted DNA and PCR-EIA increased detection to 41.2% (n = 21/51) and PapilloCheck detected 78.4% (n = 40/51). HR HPV detection by PapilloCheck was significantly higher compared with the other methods of re-analysis. Eleven samples were persistently HR HPV negative but 4 tested positive for low risk HPV. Conclusions This study identifies that up to 78% of samples with dyskaryotic cervical cytology that test negative for HPV can be found to be HPV positive on re-analysis. The reliance on a single negative HPV test result could lead to missed HPV related disease in a subset of patients, the number dependant on which HPV test is performed. The clinical significance of a false negative HPV result depends on the screening interval and how HPV testing is incorporated into screening

Direct Sequence Detection of Structured H5 Influenza Viral RNA

We describe the development of sequence-specific molecular beacons (dual-labeled DNA probes) for identification of the H5 influenza subtype, cleavage motif, and receptor specificity when hybridized directly with in vitro transcribed viral RNA (vRNA). The cloned hemagglutinin segment from a highly pathogenic H5N1 strain, A/Hanoi/30408/2005(H5N1), isolated from humans was used as template for in vitro transcription of sense-strand vRNA. The hybridization behavior of vRNA and a conserved subtype probe was characterized experimentally by varying conditions of time, temperature, and Mg2+ to optimize detection. Comparison of the hybridization rates of probe to DNA and RNA targets indicates that conformational switching of influenza RNA structure is a rate-limiting step and that the secondary structure of vRNA dominates the binding kinetics. The sensitivity and specificity of probe recognition of other H5 strains was calculated from sequence matches to the National Center for Biotechnology Information influenza database. The hybridization specificity of the subtype probes was experimentally verified with point mutations within the probe loop at five locations corresponding to the other human H5 strains. The abundance frequencies of the hemagglutinin cleavage motif and sialic acid recognition sequences were experimentally tested for H5 in all host viral species. Although the detection assay must be coupled with isothermal amplification on the chip, the new probes form the basis of a portable point-of-care diagnostic device for influenza subtyping

Global Genomic Diversity of Human Papillomavirus 6 Based on 724 Isolates and 190 Complete Genome Sequences

Human papillomavirus type 6 (HPV6) is the major etiological agent of anogenital warts and laryngeal papillomas and has been included in both the quadrivalent and nonavalent prophylactic HPV vaccines. This study investigated the global genomic diversity of HPV6, using 724 isolates and 190 complete genomes from six continents, and the association of HPV6 genomic variants with geographical location, anatomical site of infection/disease, and gender. Initially, a 2,800-bp E5a-E5b-L1-LCR fragment was sequenced from 492/530 (92.8%) HPV6-positive samples collected for this study. Among them, 130 exhibited at least one single nucleotide polymorphism (SNP), indel, or amino acid change in the E5a-E5b-L1-LCR fragment and were sequenced in full. A global alignment and maximum likelihood tree of 190 complete HPV6 genomes (130 fully sequenced in this study and 60 obtained from sequence repositories) revealed two variant lineages, A and B, and five B sublineages: B1, B2, B3, B4, and B5. HPV6 (sub)lineage-specific SNPs and a 960-bp representative region for whole-genome-based phylogenetic clustering within the L2 open reading frame were identified. Multivariate logistic regression analysis revealed that lineage B predominated globally. Sublineage B3 was more common in Africa and North and South America, and lineage A was more common in Asia. Sublineages B1 and B3 were associated with anogenital infections, indicating a potential lesion-specific predilection of some HPV6 sublineages. Females had higher odds for infection with sublineage B3 than males. In conclusion, a global HPV6 phylogenetic analysis revealed the existence of two variant lineages and five sublineages, showing some degree of ethnogeographic, gender, and/or disease predilection in their distribution.Fil: Jelen, Mateja M. University of Ljubljana; EsloveniaFil: Chen, Zigui. Albert Einstein College of Medicine. New York; Estados UnidosFil: Kocjan, Botjan J.. University of Ljubljana; EsloveniaFil: Burt, Felicity J.. University of the Free State. Bloemfontein; SudáfricaFil: Chan, Paul K.S.. Chinese University of Hong Kong; ChinaFil: Chouhy, Diego. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Biología Molecular y Celular de Rosario. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Instituto de Biología Molecular y Celular de Rosario; ArgentinaFil: Combrinck, Catharina E.. University of the Free State. Bloemfontein; SudáfricaFil: Coutlee, Francois. Hôpital Notre-Dame-Pavillon Deschamps. Montréal; CanadáFil: Estrade, Christine. Lausanne University Hospital; SuizaFil: Ferenczy, Alex. McGill University and Jewish General Hospital. Montreal; CanadáFil: Fiander, Alison. Institute of Cancer and Genetics. Cardiff; Reino UnidoFil: Franco, Eduardo L.. McGill University. Montreal; CanadáFil: Garland, Suzanne M.. The Royal Women's Hospital. Parkville; Australia. The University of Melbourne; Australia. Murdoch Children's Research Institute. Victoria; AustraliaFil: Giri, Adriana Angelica. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Biología Molecular y Celular de Rosario. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Instituto de Biología Molecular y Celular de Rosario; ArgentinaFil: González, Joaquín Víctor. Dirección Nacional de Instituto de Investigación. Administración Nacional de Laboratorio e Instituto de Salud; ArgentinaFil: Gröning, Arndt. Medizinisches Versorgungszentrum wagnerstibbe für Laboratoriumsmedizin und Pathologie GmbH. Hannover; AlemaniaFil: Heidrich, Kerstin. Medizinisches Versorgungszentrum wagnerstibbe für Laboratoriumsmedizin und Pathologie GmbH. Hannover; AlemaniaFil: Hibbitts, Sam. Institute of Cancer and Genetics. Cardiff; Reino UnidoFil: Hosnjak, Lea. University of Ljubljana; EsloveniaFil: Luk, Tommy N.M. The Chinese University of Hong Kong; China. Hong Kong Special Administrative Region; ChinaFil: Marinic, Karina. Hospital Dr. Julio C. Perrando. Chaco; ArgentinaFil: Matsukura, Toshihiko. National Institute of Health. Tokyo; JapónFil: Neumann, Anna. The University of Melbourne; Australia. Medizinisches Versorgungszentrum wagnerstibbe für Laboratoriumsmedizin und Pathologie GmbH. Hannover; AlemaniaFil: Oštrbenk, Anja. University of Ljubljana; EsloveniaFil: Picconi, Maria Alejandra. Dirección Nacional de Instituto de Investigación. Administración Nacional de Laboratorio e Instituto de Salud; ArgentinaFil: Richardson, Harriet. Queen's University. Ontario; CanadáFil: Sagadin, Martin. University of Ljubljana; EsloveniaFil: Sahli, Roland. Lausanne University Hospital; SuizaFil: Seedat, Riaz Y.. University of the Free State. Bloemfontein; SudáfricaFil: Seme, Katja. University of Ljubljana; EsloveniaFil: Severini, Alberto. Public Health Agency of Canada. Winnipeg; CanadáFil: Sinchi, Jessica L. Hospital Dr. Julio C. Perrando. Chaco; ArgentinaFil: Smahelova, Jana. Institute of Hematology and Blood Transfusion. Praga; República ChecaFil: Tabrizi, Sepehr N.. The University of Melbourne; Australia. Murdoch Children's Research Institute. Victoria; Australia. The Royal Women's Hospital. Parkville; AustraliaFil: Tachezy, Ruth. Institute of Hematology and Blood Transfusion. Praga; República ChecaFil: Tohme, Sarah. Public Health Agency of Canada. Winnipeg; CanadáFil: Uloza, Virgilijus. Lithuanian University of Health Sciences. Kaunas; LituaniaFil: Vitkauskiene, Astra. Lithuanian University of Health Sciences. Kaunas; LituaniaFil: Wong, Yong Wee. Universti Kebangsaan Malaysia. Bangi; MalasiaFil: Lepeja, Snježana Židovec. University Hospital for Infectious Diseases Dr. Fran Mihaljević. Zagreb; CroaciaFil: Burk, Robert D.. Albert Einstein College of Medicine.New York; Estados UnidosFil: Poljak, Mario. Albert Einstein College of Medicine. New York; Estados Unido