ArticleMolecular biology reports2026
Evaluating the influence of different anticoagulant blood collection tubes on the efficiency of circulating RNA extraction and Real-Time PCR quantification.
Article in Molecular biology reports, 2026. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Not yet cited in PubMed.
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Abstract
BACKGROUND AND
aimsRigorous pre-analytical standardization is a prerequisite for reliable use of circulating microRNAs and RNA as clinical biomarkers. This study evaluates the influence of four common anticoagulants (sodium citrate, Dipotassium Ethylenediaminetetraacetic Acid (K2-EDTA), Lithium heparin, and Acid Citrate Dextrose (ACD)) on isolation and quantification of miRNA and RNA from human plasma. MATERIALS AND
methodsFifteen healthy volunteers were recruited from the research staff and donated blood to study the effects of different anticoagulants on molecular tests. After Trizol extraction, we assessed RNA quantification and purity via spectrophotometry and amplification efficiency via Real-Time PCR for miR-133a, cel-miR-39 (exogenous spike-in), and U6snRNA (endogenous control).
resultsOur results confirmed that while K2-EDTA, sodium citrate and ACD provide comparable enzymatic environments for Real-time PCR, lithium heparin strongly inhibited reverse transcriptase and DNA polymerase activity, preventing successful amplification. Furthermore, specific differences in spectrophotometric purity ratios (A
conclusionsFor researchers focusing on circulating RNA and miRNA, sodium citrate, ACD and K2-EDTA should be regarded as suitable anticoagulants, with sodium citrate and ACD offering slightly better spectrophotometric purity profiles. Lithium heparin is strongly discouraged for studies involving enzymatic amplification (PCR), as the Trizol extraction method is insufficient to remove the inhibitory heparin molecules.
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