Diagnosis of genital infection caused by human papillomavirus using in situ hybridisation: the importance of the size of the biopsy specimen.

Gómez, Francisco Javier Torres; Corcuera, María Teresa; Muñoz, Elisa; M, Roldán; Alonso, María José; Abad, María del Mar; Paz, J.I.; López-Bravo, A.

doi:10.1136/jcp.48.1.57

Cited by 10 publications

(1 citation statement)

References 8 publications

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“…Typing-The specimens that were positive on screening were studied using a NISH commercial kit ('PathoGene', Enzo Diagnostics) designed for detecting the DNA of low-risk (6, ll), intermediate-risk (31, 33, 51) and high-risk (16,18) HPVs. Only the DNA sequences complementary to those used in the probes could be detected, since NISH was carried out under high stringency conditions and following the protocol described by the manufacturer of the kit.…”

Section: In Situ Hybridizationmentioning

confidence: 99%

Labelling Pattern Obtained by Non-Isotopicin Situ Hybridization as a Prognostic Factor in HPV-Associated Lesions

Gómez-Aguado

Picazo

et al. 1996

J. Pathol.

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In the study of infection of the lower female genital tract caused by human papillomavirus (HPV), one of the main concerns is the search for prognostic factors to predict the evolution of premalignant low‐ and high‐grade squamous intraepithelial lesions. This study has evaluated the prognostic usefulness of the patterns of positive reaction obtained by non‐isotopic in situ hybridization (NISH), referred to as diffuse, punctate, or mixed ‘labelling patterns’. The study examined 141 vulvar and uterine cervical biopsy specimens that were positive for HPV by a NISH screening technique and that had the following histological diagnoses: low‐grade squamous intraepithelial lesion (LSIL; n=87); high‐grade squamous intraepithelial lesion (HSIL, n=40); and squamous cell carcinoma (SCC n=14). Typing of all the specimens was carried out by NISH with DNA probes specific for HPV types 6/11 (low risk), 16/18 (high risk), and 31/33/51 (intermediate risk), and the labelling pattern observed in each specimen was recorded. Statistical analysis of the results showed that there was a significant difference in the distribution of labelling patterns, both by lesion diagnosis (P≤0·004) and by infecting viral type (P≤10−6). Lesions with a punctate or mixed pattern are considered more likely to undergo malignant evolution and consequently have a worse prognosis than lesions with a diffuse pattern.

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Section: In Situ Hybridizationmentioning

confidence: 99%

Labelling Pattern Obtained by Non-Isotopicin Situ Hybridization as a Prognostic Factor in HPV-Associated Lesions

Gómez-Aguado

Picazo

et al. 1996

J. Pathol.

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In Situ hybridization and its diagnostic applications in pathology

1997

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In situ hybridization (ISH) is a technique by which specific nucleotide sequences are identified in cells or tissue sections. These may be endogenous, bacterial or viral, DNA or RNA. On the basis of research applications, the technique is now being translated into diagnostic practice, mainly in the areas of gene expression, infection and interphase cytogenetics. Diagnostic applications are most often based on short nucleotide sequences (oligomers) labelled with non‐isotopic reporter molecules, and sites of binding may be localized by histochemical or immunohistochemical methods. The technique can be applied to routinely fixed and processed tissues; with some targets, it is even possible to obtain hybridization in autopsy material. ISH has been used to detect messenger RNA (mRNA) as a marker of gene expression, where levels of protein storage are low; for example, to confirm an endocrine tumour as the source of excess hormone production. Its application in infectious diseases has to date been mainly in viral infections, such as the typing of human papillomavirus (HPV) or the detection of Epstein–Barr virus by the presence of small nuclear RNAs (EBERs). The expression of mRNAs for histone proteins has been used to detect cells in S phase, and related methods may be applied to detect apoptotic cells. Using probes to chromosome‐specific sequences, it is possible to detect aneuploidy, and to document changes in specific chromosomes, which may have prognostic significance in some tumours, such as B‐cell chronic lymphatic leukaemia. Using sequence‐specific probes, translocations can be identified, such as the t(11;12) of Ewing's sarcoma. This review presents an outline of the technique of in situ hybridization and discusses areas of current and potential diagnostic application. © 1997 John Wiley & Sons, Ltd.

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Development of Selective Bisubstrate‐Based Inhibitors Against Protein Kinase C (PKC) Isozymes By Using Dynamic Peptide Microarrays

Poot¹,

Ameijde²,

Slijper³

et al. 2009

ChemBioChem

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Kinase inhibitors are increasingly important in drug development. Because the majority of current inhibitors target the conserved ATP-binding site, selectivity might become an important issue. This could be particularly problematic for the potential drug target protein kinase C (PKC), of which twelve isoforms with high homology exist in humans. A strategy to increase selectivity is to prepare bisubstrate-based inhibitors that target the more selective peptide-binding site in addition to the ATP-binding site. In this paper a generally applicable, rapid methodology is presented to discover such bisubstrate-based leads. Dynamic peptide microarrays were used to find peptide-binding site inhibitors. These were linked with chemoselective click chemistry to an ATP-binding site inhibitor, and this led to novel bisubstrate structures. The peptide microarrays were used to evaluate the resulting inhibitors. Thus, novel bisubstrate-based inhibitors were obtained that were both more potent and selective compared to their constituent parts. The most promising inhibitor has nanomolar affinity and selectivity towards PKCtheta amongst three isozymes.

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Diagnosis of genital infection caused by human papillomavirus using in situ hybridisation: the importance of the size of the biopsy specimen.

Cited by 10 publications

References 8 publications

Labelling Pattern Obtained by Non-Isotopicin Situ Hybridization as a Prognostic Factor in HPV-Associated Lesions

Labelling Pattern Obtained by Non-Isotopicin Situ Hybridization as a Prognostic Factor in HPV-Associated Lesions

In Situ hybridization and its diagnostic applications in pathology

Development of Selective Bisubstrate‐Based Inhibitors Against Protein Kinase C (PKC) Isozymes By Using Dynamic Peptide Microarrays

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