Comparison of the sensitivity and specificity of real-time PCR and &lt;i&gt;in situ&lt;/i&gt; hybridization in HPV16 and 18 detection in archival cervical cancer specimens

Biesaga, Beata; Szostek, Sława; Klimek, Małgorzata; Jakubowicz, Jerzy; Wysocka, Joanna

doi:10.5603/fhc.2012.0033

Cited by 11 publications

(10 citation statements)

References 25 publications

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“…Viral DNA detection was positive by real-time PCR whereas viral RNA and DNA detection were negative on histological sections by ISH for Pacific oyster spat infected with the lowest dose (L). These results could be partly explained through the higher sensitivity of real-time PCR in comparison with ISH (Biesaga et al, 2012).…”

Section: Discussionmentioning

confidence: 99%

Detection and distribution of ostreid herpesvirus 1 in experimentally infected Pacific oyster spat

Segarra

Baillon

Faury

et al. 2016

Journal of Invertebrate Pathology

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High mortality rates are reported in spat and larvae of Pacific oyster Crassostrea gigas and associated with ostreid herpesvirus 1 (OsHV-1) detection in France. Although the viral infection has been experimentally reproduced in oyster larvae and spat, little knowledge is currently available concerning the viral entry and its distribution in organs and tissues. This study compares OsHV-1 DNA and RNA detection and localization in experimentally infected oysters using two virus doses: a low dose that did not induce any mortality and a high dose inducing high mortality. Real time PCR demonstrated significant differences in terms of viral DNA amounts between the two virus doses. RNA transcripts were detected in oysters receiving the highest dose of viral suspension whereas no transcript was observed in oysters injected with the low dose. This study also allowed observing kinetics of viral DNA and RNA detection in different tissues of oyster spat. Finally, viral detection was significantly different in function of tissues (p<0.005), time (p<0.005) with an interaction between tissues and time (p<0.005) for each probe.

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Section: Discussionmentioning

confidence: 99%

Detection and distribution of ostreid herpesvirus 1 in experimentally infected Pacific oyster spat

Segarra

Baillon

Faury

et al. 2016

Journal of Invertebrate Pathology

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“…The HPV16 presence was determined on the basis of amplification of 81-bp fragment of virus E6 gene with primers (F: GAG AAC TGC AAT GTT TCA GGA CC, R: TGT ATA GTT GTT TGC AGC TCT GTG C) and TaqMan probe (6FAM-CAG GAG CGA CCC AGA AAG TTA CCA CAG TT-TAMRA), synthesized by Thermo Fisher Scientific, Waltham, USA, as previously described in detail (Biesaga et al 2012 ). In brief, amplification was carried out in a 25 µl mixture containing: 12.5 µl of Fast Universal PCR Master Mix (2 X), 100 nM of each primer, 300 nM of probe and 80 ng of DNA template.…”

Section: Methodsmentioning

confidence: 99%

Differences in the prognosis of HPV16-positive patients with squamous cell carcinoma of head and neck according to viral load and expression of P16

Biesaga

Mucha-Małecka

Janecka-Widła

et al. 2017

J Cancer Res Clin Oncol

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PurposeTo evaluate the impact of HPV16 load (VL—the number of virus genome copies per cell) and P16 expression on prognosis of patients with squamous cell carcinomas (SCCs) of head and neck (HN).Materials and methods HPV16 presence was assessed in the group of 109 patients with HNSCCs by quantitative polymerase chain reaction (qPCR). VL (assessed by qPCR) and P16 expression (evaluated by immunohistochemistry) were analysed only in the subgroup of HPV16-positive tumours. These features were correlated with 5-year overall survival (OS) and disease-free survival (DFS).ResultsHPV16 infection was found in 36 tumours (33.0%). Virus-positive patients had better OS and DFS than those without infection (P = 0.041 and 0.005). Among HPV16-positive HNSCCs, 18 (50.0%) had higher VL (median value > 6764.3 copies/cell) and 25 (73.5%) P16 over expression. The significant differences in OS and DFS (P = 0.008 and 0.004) were noticed according to VL, wherein 100% DFS was found for patients with higher VL. According to P16 expression, significant difference was found only for OS (P = 0.020). In multivariate analysis, VL (P = 0.045; HR = 2.795; CI 0.121–1.060) and the level of smoking (P = 0.023, HR = 2.253; CI 1.124–4.514) were independent factors affecting DFS of HPV16-positive patients.ConclusionOn the basis of viral load, it is possible to differentiate prognosis of patients with HPV16-positive HNSCCs. In this subgroup, viral load has stronger prognostic potential than P16 expression.

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“…Although previous studies have reported and identified endogenous retroviruses in bovine tissues (Baba et al., ; Nakaya & Miyazawa, ; Xiao, Kim, et al., ; Xiao, Park, et al., ), the current study is the first report on expression of endogenous retroviruses in pre‐implantation bovine embryo. We used RT‐qPCR technique with a very dynamic range and high sensitivity, specificity and accuracy (Benoy et al., ; Biesaga, Szostek, Klimek, Jakubowicz, & Wysocka, ; Cleusix, Lacroix, Dasen, Leo, & Le Blay, ; Tse et al., ). Our findings on the expression level of two BERVs, BERV‐K1 and BERV‐K2 envelope transcripts, reveal that these two ERVs are expressed throughout different stages of pre‐implantation development.…”

Section: Discussionmentioning

confidence: 99%

Expression of endogenous retroviruses in pre‐implantation stages of bovine embryo

Khazaee

Farzaneh

Mirshokraei

et al. 2018

Reprod Domestic Animals

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Endogenous retroviruses (ERVs) are involved in cellular proliferation, pluripotency, tissue-specific remodelling and regulation of developmental processes. These elements are transcriptionally active in mouse and human pre-implantation embryos. Empirical evidence indicates that regulatory networks involved with ERV transcripts are responsible for pluripotency and totipotency at certain stages of mouse and human pre-implantation development. Yet, the expression in pre-implantation bovine embryo remains unidentified. To determine whether two members of bovine endogenous retroviruses, BERV-K1 and BERV-K2, are expressed in the pre-implantation bovine embryo, each embryonic stage developed in vitro and was subjected to RNA release, reverse transcription and quantitative PCR. We found that BERV-K1 and BERV-K2 are expressed throughout different stages of pre-implantation development. The higher level of expression was detected in embryonic blastomeres with totipotent/pluripotent status (two-cell to 16-cell stages), while the more differentiated blastocyst stage showed significantly lower levels of ERVs expression. These findings suggest a possible role for endogenous retroviruses in the establishment of totipotent and pluripotent states in pre-implantation bovine embryo, similar to functions which have been suggested for these elements in human and mouse embryos.

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Comparison of the sensitivity and specificity of real-time PCR and <i>in situ</i> hybridization in HPV16 and 18 detection in archival cervical cancer specimens

Cited by 11 publications

References 25 publications

Detection and distribution of ostreid herpesvirus 1 in experimentally infected Pacific oyster spat

Detection and distribution of ostreid herpesvirus 1 in experimentally infected Pacific oyster spat

Differences in the prognosis of HPV16-positive patients with squamous cell carcinoma of head and neck according to viral load and expression of P16

Expression of endogenous retroviruses in pre‐implantation stages of bovine embryo

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