Multiplexed PCR genotyping of HPVs from plantaris verrucae

Lai, Jenn‐Haung; Doyle, Rebecca; Bluhm, J.M.; Johnson, James C.

doi:10.1016/j.jcv.2005.11.012

Cited by 10 publications

(5 citation statements)

References 20 publications

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“…Furthermore, in seven condylomata acuminata that were previously HPV6-positive using Low-risk Alpha-PV PCR, the presence of HPV6 was confirmed with the HPV6/11 RT-PCR and all seven samples tested HPV2/27/57-negative using the HPV2/27/57 multiplex RT-PCR ( Table 2, samples nos. [24][25][26][27][28][29][30]. Using the PCR protocols mentioned above, Alpha-PV DNA was absent in three and five samples of tested verrucae vulgares and condylomata acuminata, respectively ( , showing that the HPV2/27/57 multiplex RT-PCR is able to detect at least 10 viral copies of each targeted HPV type per a single reaction.…”

Section: Resultsmentioning

confidence: 94%

“…In contrast to previously described conventional PCRs (23)(24)(25)(26)(27)(28)(29)(30), which amplify up to 835-bp fragments of HPV DNA, the HPV2/27/57 multiplex RT-PCR targets significantly shorter HPV DNA fragments (144-157-bp), also rendering it very appropriate for HPV typing in archival tissue specimens (51). Furthermore, the majority of conventional broad-spectrum PCRs are not suitable for detecting viral targets present in low concentrations, and Sanger sequencing of PCR products hinders the identification of concurrent HPV infections.…”

Section: Discussionmentioning

confidence: 90%

“…In addition to in situ hybridization methods (20)(21)(22), several conventional broad-spectrum polymerase chain reactions (PCR)-which enable detection and differentiation of HPV types that are etiologically associated with condylomata acuminata and verrucae vulgares by subsequent laborious and time-consuming typing of PCR products using agarose gel electrophoresis, hybridization on strips/microtiter wells, and direct Sanger sequencing-have been described previously (23)(24)(25)(26)(27)(28)(29)(30)(31). Because in situ hybridization and conventional PCRs are suboptimal methods, de Koning et al (32) and Schmitt et al (33) developed broad-spectrum HPV typing bead-based xMAP Luminex suspension arrays, which are able to detect and differentiate 23 and 19 HPV types, respectively, that are most frequently found in common warts, including HPV2, HPV27, and HPV57.…”

Section: Introductionmentioning

confidence: 99%

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Development of a novel multiplex type-specific quantitative real-time PCR for detection and differentiation of infections with human papillomavirus types HPV2, HPV27, and HPV57

Hošnjak

Komloš

Kocjan

et al. 2016

Acta Dermatovenerologica Alpina Pannonica Et Adriatica

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Introduction:The present study describes the development and evaluation of the first multiplex type-specific quantitative realtime PCR (RT-PCR), enabling simple, rapid, sensitive, and specific concurrent detection and differentiation of human papillomavirus (HPV) types HPV2, 27, and 57 in a single PCR reaction. Results: The HPV2/27/57 multiplex RT-PCR with a dynamic range of seven orders of magnitude (discriminating 10 to 10 8 viral genome equivalents/reaction) has an analytical sensitivity of at least 10 viral copies of each targeted HPV type/reaction, and no cross-reactivities were observed among the included targets. All three primer/probe combinations were efficient in amplifying 500 copies of targeted DNA in a background of 10 8 , 10 7 , 500, 100, and 10 copies of non-targeted viral DNA/reaction, and the performance of the HPV2/27/57 multiplex RT-PCR was additionally not affected by the presence of background human genomic DNA. When testing DNA isolates obtained from fresh-frozen tissue specimens of various children's warts, the results of the HPV2/27/57 multiplex RT-PCR were completely in line with the results of the conventional Low-risk Alpha-PV PCR. Conclusion: The newly developed HPV2/27/57 multiplex RT-PCR is an appropriate test for use in routine clinical laboratory settings and for studies focusing on the molecular epidemiology, pathogenesis, and natural history of HPV2/27/57-related lesions.

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Section: Resultsmentioning

confidence: 94%

Section: Discussionmentioning

confidence: 90%

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Development of a novel multiplex type-specific quantitative real-time PCR for detection and differentiation of infections with human papillomavirus types HPV2, HPV27, and HPV57

Hošnjak

Komloš

Kocjan

et al. 2016

Acta Dermatovenerologica Alpina Pannonica Et Adriatica

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“…Twenty-five consecutive immunocompetent persons seeking treatment for persistent cutaneous warts in general practice (8) or outpatient dermatology department (17) were enrolled.…”

Section: Clinical Materialsmentioning

confidence: 99%

“…1 HPV types belonging to four of those genera, i.e., Alphapapillomavirus, Gammapapillomavirus, Mupapillomavirus and Nupapillomavirus have been associated with cutaneous warts, mainly with foot and hand warts. [2][3][4][5][6][7][8][9][10][11][12][13] Reliable detection and sampling techniques are mandatory to be able to study the epidemiology of these HPV infections.…”

Section: Introductionmentioning

confidence: 99%