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

Autor(es):Mostrar menos -

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

Afiliação do(s) autor(es)

^[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

• Tipo de documento: Artigo Científico
• Fonte: GENE REPORTS; v. 7, p. 98-105, JUN 2017.
• Citações Web of Science: 3

Resumo

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)

• Processo FAPESP: 13/15879-8 - Infecção por Toxoplasma gondii: caracterização sorológica e molecular em pacientes atendidos pelo SUS
• Beneficiário: Fernando Henrique Antunes Murata
• Modalidade de apoio: Bolsas no Brasil - Mestrado
• Processo FAPESP: 13/05811-7 - Diagnóstico de doenças infecciosas emergentes e reemergentes em amostras de órgãos e tecidos
• Beneficiário: Roosecelis Brasil Martines
• Modalidade de apoio: Bolsas no Exterior - Pesquisa
• Processo FAPESP: 13/25650-8 - Genes HLA de classe II (DRB1 e DQB1) como fatores de risco para a toxoplasmose ocular
• Beneficiário: Luiz Carlos de Mattos
• Modalidade de apoio: Auxílio à Pesquisa - Regular
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• Beneficiário: Mariana Previato
• Modalidade de apoio: Bolsas no Brasil - Iniciação Científica
• Processo FAPESP: 14/09496-1 - Novas estratégias diagnósticas empregadas na toxoplasmose sintomática
• Beneficiário: Vera Lúcia Pereira Chioccola
• Modalidade de apoio: Auxílio à Pesquisa - Regular
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• Beneficiário: Daise Damaris Carnietto de Hippólito
• Modalidade de apoio: Bolsas no Brasil - Mestrado