Identification and validation of reference genes of circulating microRNAs for use as control in gestational toxoplasmosis

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Pereira, Ingrid de Siqueira ; da Cruz, Allecineia Bispo ; Maia, Marta Marques ; Carneiro, Francieli Marinho ; Gava, Ricardo ; Spegiorin, Ligia Cosentino Junqueira Franco ; Branda, Cinara Cassia ; Truzzi, Ingrid Gomes de Campos ; de Faria Junior, Geraldo Magela ; de Mattos, Luiz Carlos ; Pereira-Chioccola, Vera Lucia ; Meira-Strejevitch, Cristina Silva

• Document type: Journal article
• Source: Molecular and Biochemical Parasitology; v. 256, p. 7-pg., 2023-12-01.

Abstract

Toxoplasmosis causes serious harm to the fetus, as tachyzoite dissemination, during pregnancy in women developing the primoinfection. The microRNAs (miRNAs) are small non-coding RNAs, which have regulatory roles in cells by silencing messenger RNA. Circulating miRNA are promising biomarkers for diagnosis and prognosis of numerous diseases. The miRNAs levels are estimated by quantitative real-time PCR (qPCR), however, the relative quantification of each miRNA expression requires proper normalization methods using endogenous miRNAs as control. This study analyzed the expression of three endogenous miRNAs (miR484, miR -423-3p and miR-26b-5p) for use as normalizers in future studies of target miRNAs for gestational toxoplasmosis (GT). A total of 32 plasma samples were used in all assays divided in 21 from women with GT and 11 from healthy women. The stability of each endogenous miRNA was evaluated by the algorithm methods RefFinder that included GeNorm, Normfinder, BestKeeper and comparative delta-CT programs. The miR-484 was the most stably gene, and equivalently expressed in GT and NC groups. These results contribute to future studies of target miRNAs in clinical samples of women with gestational toxoplasmosis. (AU)

• FAPESP's process: 18/09448-8 - Expression of miRNAs in ocular Toxoplasmosis
• Grantee: Geraldo Magela de Faria Junior
• Support Opportunities: Scholarships in Brazil - Doctorate
• FAPESP's process: 20/14783-0 - MicroRNAs e their mechanisms of gene regulation in symptomatic toxoplasmosis
• Grantee: Cristina da Silva Meira Strejevitch
• Support Opportunities: Regular Research Grants