Detection and genotyping of human papillomavirus by real-time PCR assay

Moreau, F.; Fetouchi, Rachid; Micalessi, I.; Brejeon, Valérie; Bacon, Nathalie; Jannes, Geert; Pendeven, Catherine Le; Lekbaby, Bouchra; Kremsdorf, Dina; Guily, Jean Lacau Saint; Soussan, Patrick Ben

doi:10.1016/j.jcv.2012.11.003

Cited by 19 publications

(10 citation statements)

References 20 publications

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“…In some cases we also calculated the difference in cycle time (Ct) between the 18sRNA gene and viral DNA (ΔCt). Fold change (2 ΔCt ) demonstrates the relative viral DNA load in each sample as described previously 23 , 39 .…”

Section: Methodsmentioning

confidence: 99%

Mouse papillomavirus infection persists in mucosal tissues of an immunocompetent mouse strain and progresses to cancer

Cladel

Budgeon

Balogh

et al. 2017

Sci Rep

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Mouse papillomavirus has shown broad tissue tropism in nude mice. Previous studies have tested cutaneous infections in different immunocompromised and immunocompetent mouse strains. In the current study, we examined mucosal infection in several immunocompetent and immunocompromised mouse strains. Viral DNA was monitored periodically by Q-PCR of lavage samples. Immunohistochemistry and in situ hybridization were used to determine viral capsid protein and viral DNA respectively. All athymic nude mouse strains showed active infections at both cutaneous and mucosal sites. Interestingly, NOD/SCID mice, which have a deficiency in T, B, and NK cells, showed minimal disease at cutaneous sites but developed persistent infection at the mucosal sites including those of the anogenital region and the oral cavity. Three strains of immunocompetent mice supported mucosal infections. Infections of the lower genital tract in heterozygous (immunocompetent) mice of the NU/J strain progressed to high grade dysplasia and to carcinoma in situ. Anti-MmuPV1 neutralizing antibodies were detected in the sera of all immunocompetent animals. Our findings demonstrate that the mucosae may be the preferred sites for this virus in mice. The mouse model is expected to be a valuable model for the study of mucosal papillomavirus disease, progression, and host immune control.

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Section: Methodsmentioning

confidence: 99%

Mouse papillomavirus infection persists in mucosal tissues of an immunocompetent mouse strain and progresses to cancer

Cladel

Budgeon

Balogh

et al. 2017

Sci Rep

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“…To select which method should be applied for DNA extraction, we used QiaAmp ® DNA mini Kit (Qiagen, Germany) ( 16 ), phenol–chloroform extraction, guanidinium thiocyanate extraction ( 17 ), and ammonium acetate extraction ( 18 ). The best results were obtained by the ammonium acetate method.…”

Section: Methodsmentioning

confidence: 99%

PCR-RFLP assay as an option for primary HPV test

Golfetto

Alves

Martins

et al. 2018

Braz J Med Biol Res

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Persistent human papillomavirus (HPV) infection is an essential factor of cervical cancer. This study evaluated the analytical performance of restriction fragment length polymorphism polymerase chain reaction (PCR-RFLP) assay compared to PapilloCheck® microarray to identify human papilloma virus (HPV) in cervical cells. Three hundred and twenty-five women were analyzed. One sample was used for conventional cytology and another sample was collected using BD SurePath™ kit for HPV tests. Eighty samples (24.6%) were positive for HPV gene by PCR-Multiplex and were then submitted to PCR-RFLP and PapilloCheck® microarray. There was a genotyping agreement in 71.25% (57/80) on at least one HPV type between PCR-RFLP and PapilloCheck® microarray. In 22 samples (27.5%), the results were discordant and those samples were additionally analyzed by DNA sequencing. HPV 16 was the most prevalent HPV type found in both methods, followed by HPVs 53, 68, 18, 39, and 66 using PCR-RFLP analysis, and HPVs 39, 53, 68, 56, 31, and 66 using PapilloCheck® microarray. In the present study, a perfect agreement using Cohen's kappa (κ) was found in HPV 33 and 58 (κ=1), very good for HPV 51, and good for types 16, 18, 53, 59, 66, 68, 70, and 73. PCR-RFLP analysis identified only 25% (20/80) HPV coinfection, and PapilloCheck® microarray found 62.5% (50/80). Our Cohen's kappa results indicate that our in-house HPV genotyping testing (PCR-RFLP analysis) could be applied as a primary HPV test screening, especially in low income countries. If multiple HPV types are found in this primary test, a more descriptive test, such as PapilloCheck® microarray, could be performed.

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“…Papillomaviruses are small nonenveloped icosahedral viruses that carry a circular double-stranded DNA (dsDNA) genome decorated with host-derived histones. Recently, it has been suggested that individual warts can sometimes be simultaneously coinfected with multiple types of papillomaviruses (2). …”

Section: Genome Announcementmentioning

confidence: 99%