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(Reference retrieved automatically from Web of Science through information on FAPESP grant and its corresponding number as mentioned in the publication by the authors.)

Reference genes for studies in infectious parasitic diseases in five types of human tissues

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Meira-Strejevitch, Cristina Silva [1] ; Pereira-Chioccola, Vera Lucia [1] ; Maia, Marta Marques [1] ; Carnietto de Hippolito, Daise Damaris [1] ; Wang, Hui-Tzu Lin [2] ; Motoie, Gabriela [1] ; de Souza Gomes, Aparecida Helena [3] ; Kanamura, Cristina Takami [4] ; Martines, Roosecelis Brasil [4] ; Brandao de Mattos, Cinara Cassia [5] ; Frederico, Fabio Batista [6] ; de Mattos, Luiz Carlos [5] ; Grp, FAMERP Toxoplasma Res
Total Authors: 13
Affiliation:
[1] Adolfo Lutz Inst, Ctr Parasitol & Micol, Ave Dr Arnaldo 351, 8 Andar, BR-01246902 Sao Paulo, SP - Brazil
[2] Inst Dante Pazzanese Cardiol, Lab Invest Mol Cardiol, Sao Paulo - Brazil
[3] Adolfo Lutz Inst, Lab Reg Sorocaba, Sorocaba - Brazil
[4] Adolfo Lutz Inst, Ctr Patol, Sao Paulo - Brazil
[5] Fac Med Sao Jose do Rio Preto, Lab Imunogenet, Sao Jose Do Rio Preto - Brazil
[6] Fundacao Fac Reg Med Hosp Base, Ambulatorio Oftalmol, Sao Jose Do Rio Preto - Brazil
Total Affiliations: 6
Document type: Journal article
Source: GENE REPORTS; v. 7, p. 98-105, JUN 2017.
Web of Science Citations: 3
Abstract

Gene expression analyses based onmessenger RNA (mRNA) expression require accurate data normalization. When using endogenous reference genes, these should be carefully validated. Validated reference genes vary greatly depending on tissue, cell subsets and experimental context. This study was aimed to identify reference genes that have more stable mRNA levels among individuals in peripheral blood mononuclear cells (PBMC); fresh skin biopsies; lung and brain autopsies as well as, skin biopsies formalin-fixed paraffin-embedded (FFPE). Therefore, 6 endogenous reference genes were evaluated by quantitative real-time polymerase chain reaction: 18S rRNA, glyceraldehyde-3-phosphate dehydrogenase (GAPDH), TATA box-binding protein (TBP), beta-2-microbolin (B2M), ubiquitin C (UBC) and mitochondrially encoded ATP synthase 6 (MT-ATP6). Furthermore, validation of their stabilities and performance as reference genes was determined by geNorm and NormFinder programs. The results show that the most stable genes for PBMC and fresh skin biopsies were TBP and UBC; in FFPE lung autopsies and skin biopsies were GAPDH and B2M; and in FFPE brain autopsies were GAPDH and UBC. In addition, 18S rRNA was the least stable of all genes analyzed. These data concluded that even genes constitutively expressed have transcript level variations in different tissues as well as storage and experimental conditions. These observations suggest that suitable reference genes should be selected for normalization of gene expression data analysis. (C) 2017 Elsevier Inc. All rights reserved. (AU)

FAPESP's process: 13/10050-5 - Serological, molecular and images in patients with active lesions or acute relapse of ocular infection by Toxoplasma gondii: a study follow-up
Grantee:Mariana Previato
Support type: Scholarships in Brazil - Scientific Initiation
FAPESP's process: 14/09496-1 - New diagnostic strategies employed in symptomatic toxoplasmosis
Grantee:Vera Lúcia Pereira Chioccola
Support type: Regular Research Grants
FAPESP's process: 13/15879-8 - Toxoplasma gondii infection: serological and molecular characterization in patients attended in public health service (SUS)
Grantee:Fernando Henrique Antunes Murata
Support type: Scholarships in Brazil - Master
FAPESP's process: 15/04803-6 - Cytokine expression in the modulation of cutaneous leishmaniasis and its interaction with clinical, epidemiological and laboratory findings
Grantee:Daise Damaris Carnietto de Hippólito
Support type: Scholarships in Brazil - Master
FAPESP's process: 13/05811-7 - Emerging and reemerging infectious diseases diagnosis in samples of organs and tissues
Grantee:Roosecelis Brasil Martines
Support type: Scholarships abroad - Research
FAPESP's process: 13/25650-8 - Class II HLA genes (DRB1 and DQB1) as risck factors for ocular toxoplasmosis
Grantee:Luiz Carlos de Mattos
Support type: Regular Research Grants