Cancer Diagnosis Program | Biorepositories and Biospecimen Research Branch

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Improved Discrimination of Patients with Breast Cancer from Healthy Controls Using Paper-Based microRNA Expression Profiling of Plasma, Following Precipitation.

Author(s): Tan KML, Tay SP, Chua HW, Sufyan Bin Masroni M, Tucker S, Koay ES, Leong SM

Publication: Clin Chem, 2017, Vol. 63, Page 1899-1901

PubMed ID: 28923847 PubMed Review Paper? No

Purpose of Paper

Conclusion of Paper

Studies

1. Study Purpose

   This study investigated if extracting miRNA from plasma precipitates rather than plasma improves discrimination between patients with breast cancer and healthy controls. Peripheral blood from 9 healthy controls and 9 patients with breast cancer was centrifuged at 3000 x g for 10 min to obtain plasma. Plasma was stored at -80˚C and visually inspected to confirm the absence of hemolysis. RNA was extracted directly from plasma using the FTA Elute MicroCard. To investigate the effect of precipitation, Total Exosome Isolation reagent was added to plasma and the specimen was vortexed and centrifuged at 10000 x g for 10 min. The pellet was resuspended in PBS, spotted on an FTA Elute MicroCard, and then the card was washed in 70% ethanol. RNA was eluted with DEPC water and miRNAs were quantified using the real-time PCR MiRXES IDEAL miRNA assays. Values were normalized to the mean quantification cycle (Cq) value for all miRNA.

   Summary of Findings:

   Plasma precipitates from breast cancer patients had significantly higher levels of miR-16 (P=0.0020), miR-106b (P=0.0135), and miR-486-5p (P=0.0047) than precipitates from healthy controls. Plasma showed more overlap in Cq values between breast cancer patients and controls than what was observed from precipitates, but plasma from breast cancer patients had significantly higher levels of miR-16 (P=0.0244), miR-106b (P=0.0424), and miR-486-5p (P=0.0173) than healthy controls. Principle component analysis confirmed better discrimination between breast cancer patients and controls in precipitates than plasma. The precipitates had a larger area under the receiver operator curves (AUROC) than plasma for all four miRNAs examined.

   Biospecimens

   • Bodily Fluid - Plasma

   Preservative Types

   • Frozen

   Diagnoses:

   • Neoplastic - Carcinoma
   • Normal

   Platform:

   Analyte	Technology Platform
   RNA	Real-time qRT-PCR

   Pre-analytical Factors:

   Classification	Pre-analytical Factor	Value(s)
   Preaquisition	Diagnosis/ patient condition	Breast cancer Healthy
   Real-time qRT-PCR Specific	Targeted nucleic acid	miR-16 miR-106b miR-222 miR-486-5p
   Biospecimen Aliquots and Components	Blood and blood products	Plasma Plasma precipitates

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