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1.
J Immunol Methods ; 465: 13-19, 2019 02.
Artículo en Inglés | MEDLINE | ID: mdl-30496732

RESUMEN

BACKGROUND: Uncontrolled preanalytical variables can reduce the accuracy and reproducibility of downstream analytical results from peripheral blood mononuclear cells (PBMCs). METHODS: PBMCs were isolated from EDTA and citrate-anticoagulated blood samples, obtained from healthy subjects and patients with inflammatory and infectious conditions. PBMC-derived RNA samples were examined for gene expression changes induced by extended blood pre-centrifugation delays at 4 °C and RT. We used Taqman RTqPCR to evaluate the combination of two target genes for their "diagnostic performance" in identifying EDTA and citrate-anticoagulated PBMC samples with extended pre-centrifugation times. RESULTS: We established the PBMC preanalytical score, a gene expression metric to asses the PBMC quality related to the pre-centrifugation delay at room temperature for different anticoagulants. The PBMC preanalytical score measurement can identify: CONCLUSION: The proposed PBMC preanalytical score may enable objective PBMC sample qualification for downstream applications, which may be influenced by blood precentrifugation delays.


Asunto(s)
Proteínas de Unión al Calcio/biosíntesis , Regulación de la Expresión Génica , Interleucina-8/biosíntesis , Leucocitos Mononucleares/metabolismo , alfa-Manosidasa/biosíntesis , Adulto , Anciano , Femenino , Humanos , Leucocitos Mononucleares/citología , Masculino , Persona de Mediana Edad
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