Validation of chromogenic in situ hybridization reactions for DNA and RNA detection in formalin-fixed paraffin-embedded tissue

Detalhes bibliográficos
Autor(a) principal: Monteiro,Raquel L.
Data de Publicação: 2019
Outros Autores: Damaceno,Daniela S., Kimura,Lidia M., Cirqueira,Cinthya S., Guerra,Juliana M., Araújo,Leonardo José T.
Tipo de documento: Artigo
Idioma: eng
Título da fonte: Jornal Brasileiro de Patologia e Medicina Laboratorial (Online)
Texto Completo: http://old.scielo.br/scielo.php?script=sci_arttext&pid=S1676-24442019000100057
Resumo: ABSTRACT Introduction: Chromogenic in situ hybridization (CISH) is used alternatively to the traditional immunohistochemical methods for the diagnosis of infectious diseases in formalin-fixed paraffin-embedded samples, since it presents high sensitivity and specificity. This type of sample undergoes several chemical modifications during histological processing, and both poor and excessive fixation can impair sample quality, making it difficult to obtain good results. In CISH, it is common to use positive samples as quality control for the reactions; however, this practice does not provide any information regarding the preservation of the genetic material, nor does it avoid false-negative results. Objective: The objective of this study was to validate the deoxyribonucleic acid (DNA) (+) and (-), and ribonucleic acid (RNA) (+) and (-) control probes to be used as quality control for the samples, evaluating preservation of the genetic material. Materials and methods: Twelve histological sections were used (in quadruplicate, n = 48), prepared from a pool of tissues without microscopic changes related to infectious and/or inflammatory processes. The CISH protocol was conducted according to the manufacturer’s instructions, standardized under the conditions of our laboratory, using commercial DNA and RNA probes chemically linked to digoxigenin. Results and conclusion: Our results were very satisfactory, showing high reproducibility, accuracy, sensitivity and analytical specificity, high predictive values for positive and negative assays and with zero ratio of false-positive and false-negative results, allowing the validation of this reaction.
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spelling Validation of chromogenic in situ hybridization reactions for DNA and RNA detection in formalin-fixed paraffin-embedded tissuein situ hybridizationquality controlmolecular diagnostic techniquesABSTRACT Introduction: Chromogenic in situ hybridization (CISH) is used alternatively to the traditional immunohistochemical methods for the diagnosis of infectious diseases in formalin-fixed paraffin-embedded samples, since it presents high sensitivity and specificity. This type of sample undergoes several chemical modifications during histological processing, and both poor and excessive fixation can impair sample quality, making it difficult to obtain good results. In CISH, it is common to use positive samples as quality control for the reactions; however, this practice does not provide any information regarding the preservation of the genetic material, nor does it avoid false-negative results. Objective: The objective of this study was to validate the deoxyribonucleic acid (DNA) (+) and (-), and ribonucleic acid (RNA) (+) and (-) control probes to be used as quality control for the samples, evaluating preservation of the genetic material. Materials and methods: Twelve histological sections were used (in quadruplicate, n = 48), prepared from a pool of tissues without microscopic changes related to infectious and/or inflammatory processes. The CISH protocol was conducted according to the manufacturer’s instructions, standardized under the conditions of our laboratory, using commercial DNA and RNA probes chemically linked to digoxigenin. Results and conclusion: Our results were very satisfactory, showing high reproducibility, accuracy, sensitivity and analytical specificity, high predictive values for positive and negative assays and with zero ratio of false-positive and false-negative results, allowing the validation of this reaction.Sociedade Brasileira de Patologia Clínica2019-02-01info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersiontext/htmlhttp://old.scielo.br/scielo.php?script=sci_arttext&pid=S1676-24442019000100057Jornal Brasileiro de Patologia e Medicina Laboratorial v.55 n.1 2019reponame:Jornal Brasileiro de Patologia e Medicina Laboratorial (Online)instname:Sociedade Brasileira de Patologia (SBP)instacron:SBP10.5935/1676-2444.20190008info:eu-repo/semantics/openAccessMonteiro,Raquel L.Damaceno,Daniela S.Kimura,Lidia M.Cirqueira,Cinthya S.Guerra,Juliana M.Araújo,Leonardo José T.eng2019-05-08T00:00:00Zoai:scielo:S1676-24442019000100057Revistahttp://www.scielo.br/jbpmlhttps://old.scielo.br/oai/scielo-oai.php||jbpml@sbpc.org.br1678-47741676-2444opendoar:2019-05-08T00:00Jornal Brasileiro de Patologia e Medicina Laboratorial (Online) - Sociedade Brasileira de Patologia (SBP)false
dc.title.none.fl_str_mv Validation of chromogenic in situ hybridization reactions for DNA and RNA detection in formalin-fixed paraffin-embedded tissue
title Validation of chromogenic in situ hybridization reactions for DNA and RNA detection in formalin-fixed paraffin-embedded tissue
spellingShingle Validation of chromogenic in situ hybridization reactions for DNA and RNA detection in formalin-fixed paraffin-embedded tissue
Monteiro,Raquel L.
in situ hybridization
quality control
molecular diagnostic techniques
title_short Validation of chromogenic in situ hybridization reactions for DNA and RNA detection in formalin-fixed paraffin-embedded tissue
title_full Validation of chromogenic in situ hybridization reactions for DNA and RNA detection in formalin-fixed paraffin-embedded tissue
title_fullStr Validation of chromogenic in situ hybridization reactions for DNA and RNA detection in formalin-fixed paraffin-embedded tissue
title_full_unstemmed Validation of chromogenic in situ hybridization reactions for DNA and RNA detection in formalin-fixed paraffin-embedded tissue
title_sort Validation of chromogenic in situ hybridization reactions for DNA and RNA detection in formalin-fixed paraffin-embedded tissue
author Monteiro,Raquel L.
author_facet Monteiro,Raquel L.
Damaceno,Daniela S.
Kimura,Lidia M.
Cirqueira,Cinthya S.
Guerra,Juliana M.
Araújo,Leonardo José T.
author_role author
author2 Damaceno,Daniela S.
Kimura,Lidia M.
Cirqueira,Cinthya S.
Guerra,Juliana M.
Araújo,Leonardo José T.
author2_role author
author
author
author
author
dc.contributor.author.fl_str_mv Monteiro,Raquel L.
Damaceno,Daniela S.
Kimura,Lidia M.
Cirqueira,Cinthya S.
Guerra,Juliana M.
Araújo,Leonardo José T.
dc.subject.por.fl_str_mv in situ hybridization
quality control
molecular diagnostic techniques
topic in situ hybridization
quality control
molecular diagnostic techniques
description ABSTRACT Introduction: Chromogenic in situ hybridization (CISH) is used alternatively to the traditional immunohistochemical methods for the diagnosis of infectious diseases in formalin-fixed paraffin-embedded samples, since it presents high sensitivity and specificity. This type of sample undergoes several chemical modifications during histological processing, and both poor and excessive fixation can impair sample quality, making it difficult to obtain good results. In CISH, it is common to use positive samples as quality control for the reactions; however, this practice does not provide any information regarding the preservation of the genetic material, nor does it avoid false-negative results. Objective: The objective of this study was to validate the deoxyribonucleic acid (DNA) (+) and (-), and ribonucleic acid (RNA) (+) and (-) control probes to be used as quality control for the samples, evaluating preservation of the genetic material. Materials and methods: Twelve histological sections were used (in quadruplicate, n = 48), prepared from a pool of tissues without microscopic changes related to infectious and/or inflammatory processes. The CISH protocol was conducted according to the manufacturer’s instructions, standardized under the conditions of our laboratory, using commercial DNA and RNA probes chemically linked to digoxigenin. Results and conclusion: Our results were very satisfactory, showing high reproducibility, accuracy, sensitivity and analytical specificity, high predictive values for positive and negative assays and with zero ratio of false-positive and false-negative results, allowing the validation of this reaction.
publishDate 2019
dc.date.none.fl_str_mv 2019-02-01
dc.type.driver.fl_str_mv info:eu-repo/semantics/article
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Sociedade Brasileira de Patologia Clínica
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Sociedade Brasileira de Patologia Clínica
dc.source.none.fl_str_mv Jornal Brasileiro de Patologia e Medicina Laboratorial v.55 n.1 2019
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