A Novel Assay for the Identification of NOTCH1 PEST Domain Mutations in Chronic Lymphocytic Leukemia

Aims. To develop a fast and robust DNA-based assay to detect insertions and deletions mutations in exon 34 that encodes the PEST domain of NOTCH1 in order to evaluate patients with chronic lymphocytic leukemia (CLL). Methods. We designed a multiplexed allele-specific polymerase chain reaction (PCR)...

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Main Authors: Paulo Vidal Campregher, Roberta Cardoso Petroni, Nair Hideko Muto, Roberta Sitnik, Flavia Pereira de Carvalho, Nydia Strachman Bacal, Elvira Deolinda Rodrigues Pereira Velloso, Gislaine Borba Oliveira, João Renato Rebello Pinho, Davi Coe Torres, Marcela Braga Mansur, Rocio Hassan, Irene Gyongyvér Heidemarie Lorand-Metze, Carlos Sérgio Chiattone, Nelson Hamerschlak, Cristovão Luis Pitangueira Mangueira
Format: Article
Language:English
Published: Hindawi Limited 2016-01-01
Series:BioMed Research International
Online Access:http://dx.doi.org/10.1155/2016/4247908
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spelling doaj-dbbc570dc4004022b3741a63fd6b55962020-11-24T22:18:17ZengHindawi LimitedBioMed Research International2314-61332314-61412016-01-01201610.1155/2016/42479084247908A Novel Assay for the Identification of NOTCH1 PEST Domain Mutations in Chronic Lymphocytic LeukemiaPaulo Vidal Campregher0Roberta Cardoso Petroni1Nair Hideko Muto2Roberta Sitnik3Flavia Pereira de Carvalho4Nydia Strachman Bacal5Elvira Deolinda Rodrigues Pereira Velloso6Gislaine Borba Oliveira7João Renato Rebello Pinho8Davi Coe Torres9Marcela Braga Mansur10Rocio Hassan11Irene Gyongyvér Heidemarie Lorand-Metze12Carlos Sérgio Chiattone13Nelson Hamerschlak14Cristovão Luis Pitangueira Mangueira15Departments of Hematology and Clinical Pathology, and Research Institute, Hospital Israelita Albert Einstein, São Paulo, SP, BrazilDepartment of Clinical Pathology, Hospital Israelita Albert Einstein, São Paulo, SP, BrazilDepartment of Clinical Pathology, Hospital Israelita Albert Einstein, São Paulo, SP, BrazilDepartment of Clinical Pathology, Hospital Israelita Albert Einstein, São Paulo, SP, BrazilDepartment of Clinical Pathology, Hospital Israelita Albert Einstein, São Paulo, SP, BrazilDepartment of Clinical Pathology, Hospital Israelita Albert Einstein, São Paulo, SP, BrazilHematology Service, Hospital das Clínicas, Faculdade de Medicina da Universidade de São Paulo São Paulo, SP, BrazilHemocentro, Faculdade de Medicina da Universidade Estadual de Campinas (Hemocentro-Unicamp), Campinas, SP, BrazilDepartment of Clinical Pathology, Hospital Israelita Albert Einstein, São Paulo, SP, BrazilBone Marrow Transplantation Center, Instituto Nacional de Câncer (INCA), Rio de Janeiro, RJ, BrazilPaediatric Haematology-Oncology Program, Instituto Nacional de Câncer (INCA), Rio de Janeiro, RJ, BrazilBone Marrow Transplantation Center, Instituto Nacional de Câncer (INCA), Rio de Janeiro, RJ, BrazilHemocentro, Faculdade de Medicina da Universidade Estadual de Campinas (Hemocentro-Unicamp), Campinas, SP, BrazilDepartment of Hematology, Faculdade de Medicina da Santa Casa de Misericórdia de São Paulo, São Paulo, SP, BrazilDepartment of Hematology, Hospital Israelita Albert Einstein, São Paulo, SP, BrazilDepartment of Clinical Pathology, Hospital Israelita Albert Einstein, São Paulo, SP, BrazilAims. To develop a fast and robust DNA-based assay to detect insertions and deletions mutations in exon 34 that encodes the PEST domain of NOTCH1 in order to evaluate patients with chronic lymphocytic leukemia (CLL). Methods. We designed a multiplexed allele-specific polymerase chain reaction (PCR) combined with a fragment analysis assay to detect specifically the mutation c.7544_7545delCT and possibly other insertions and deletions in exon 34 of NOTCH1. Results. We evaluated our assay in peripheral blood samples from two cohorts of patients with CLL. The frequency of NOTCH1 mutations was 8.4% in the first cohort of 71 unselected CLL patients. We then evaluated a second cohort of 26 CLL patients with known cytogenetic abnormalities that were enriched for patients with trisomy 12. NOTCH1 mutations were detected in 43.7% of the patients with trisomy 12. Conclusions. We have developed a fast and robust assay combining allele-specific PCR and fragment analysis able to detect NOTCH1 PEST domain insertions and deletions.http://dx.doi.org/10.1155/2016/4247908
collection DOAJ
language English
format Article
sources DOAJ
author Paulo Vidal Campregher
Roberta Cardoso Petroni
Nair Hideko Muto
Roberta Sitnik
Flavia Pereira de Carvalho
Nydia Strachman Bacal
Elvira Deolinda Rodrigues Pereira Velloso
Gislaine Borba Oliveira
João Renato Rebello Pinho
Davi Coe Torres
Marcela Braga Mansur
Rocio Hassan
Irene Gyongyvér Heidemarie Lorand-Metze
Carlos Sérgio Chiattone
Nelson Hamerschlak
Cristovão Luis Pitangueira Mangueira
spellingShingle Paulo Vidal Campregher
Roberta Cardoso Petroni
Nair Hideko Muto
Roberta Sitnik
Flavia Pereira de Carvalho
Nydia Strachman Bacal
Elvira Deolinda Rodrigues Pereira Velloso
Gislaine Borba Oliveira
João Renato Rebello Pinho
Davi Coe Torres
Marcela Braga Mansur
Rocio Hassan
Irene Gyongyvér Heidemarie Lorand-Metze
Carlos Sérgio Chiattone
Nelson Hamerschlak
Cristovão Luis Pitangueira Mangueira
A Novel Assay for the Identification of NOTCH1 PEST Domain Mutations in Chronic Lymphocytic Leukemia
BioMed Research International
author_facet Paulo Vidal Campregher
Roberta Cardoso Petroni
Nair Hideko Muto
Roberta Sitnik
Flavia Pereira de Carvalho
Nydia Strachman Bacal
Elvira Deolinda Rodrigues Pereira Velloso
Gislaine Borba Oliveira
João Renato Rebello Pinho
Davi Coe Torres
Marcela Braga Mansur
Rocio Hassan
Irene Gyongyvér Heidemarie Lorand-Metze
Carlos Sérgio Chiattone
Nelson Hamerschlak
Cristovão Luis Pitangueira Mangueira
author_sort Paulo Vidal Campregher
title A Novel Assay for the Identification of NOTCH1 PEST Domain Mutations in Chronic Lymphocytic Leukemia
title_short A Novel Assay for the Identification of NOTCH1 PEST Domain Mutations in Chronic Lymphocytic Leukemia
title_full A Novel Assay for the Identification of NOTCH1 PEST Domain Mutations in Chronic Lymphocytic Leukemia
title_fullStr A Novel Assay for the Identification of NOTCH1 PEST Domain Mutations in Chronic Lymphocytic Leukemia
title_full_unstemmed A Novel Assay for the Identification of NOTCH1 PEST Domain Mutations in Chronic Lymphocytic Leukemia
title_sort novel assay for the identification of notch1 pest domain mutations in chronic lymphocytic leukemia
publisher Hindawi Limited
series BioMed Research International
issn 2314-6133
2314-6141
publishDate 2016-01-01
description Aims. To develop a fast and robust DNA-based assay to detect insertions and deletions mutations in exon 34 that encodes the PEST domain of NOTCH1 in order to evaluate patients with chronic lymphocytic leukemia (CLL). Methods. We designed a multiplexed allele-specific polymerase chain reaction (PCR) combined with a fragment analysis assay to detect specifically the mutation c.7544_7545delCT and possibly other insertions and deletions in exon 34 of NOTCH1. Results. We evaluated our assay in peripheral blood samples from two cohorts of patients with CLL. The frequency of NOTCH1 mutations was 8.4% in the first cohort of 71 unselected CLL patients. We then evaluated a second cohort of 26 CLL patients with known cytogenetic abnormalities that were enriched for patients with trisomy 12. NOTCH1 mutations were detected in 43.7% of the patients with trisomy 12. Conclusions. We have developed a fast and robust assay combining allele-specific PCR and fragment analysis able to detect NOTCH1 PEST domain insertions and deletions.
url http://dx.doi.org/10.1155/2016/4247908
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