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The Effect of Blood Clots on the Quality of RNA Extracted from PAXgene Blood RNA Tubes.

Author(s): Tang R, Zhu L, Zhu P, Yin R, Zheng C

Publication: Biopreserv Biobank, 2023, Vol. , Page

PubMed ID: 37540078 PubMed Review Paper? No

Purpose of Paper

Conclusion of Paper

Studies

1. Study Purpose

   This paper compared the yield, purity, and integrity of RNA extracted from patient-matched clotted and non-clotted blood that was collected in PAXgene Blood RNA tubes and between RNA extracted from specimens with small and large clots.  Fasting blood was recollected in PAXgene RNA tubes within 2 days from 15 patients who had a clotted blood specimen (diagnosis not specified). The paired clotted and new non-clotted specimens were stored at -80°C for 1-5 months. Clotted specimens were classified based on clot thickness as small if the maximum clot diameter was < 0.5 cm (4 specimens) or large if the diameter was ≥ 0.5 cm (11 specimens). Before RNA extraction with the PAXgene Blood RNA Kit, blood was thawed at room temperature for 2 h and then inverted 10 times.  RNA yield and purity were quantified by spectrophotometer. RNA integrity was analyzed using the Eukaryote Total RNA 6000 Nano LabChip Kit and the Eukaryote Total RNA Nano Assay on a Bioanalyzer.

   Summary of Findings:

   RNA yield and purity (OD 260/280 and OD 260/230) were comparable when RNA was extracted from clotted and non-clotted specimens. However, the median RIN was lower in RNA extracted from clotted than non-clotted blood (8.20 versus 7.20, P<0.05). RNA extracted from specimens with large clots (maximum diameter ≥0.5 cm) had a lower RIN than RNA extracted from specimens with small clots (maximum diameter < 0.5 cm) (6.9 versus 8.25, P<0.0001), but RNA yield and purity were similar when RNA was extracted from specimens with small and large clots. Importantly, while RNA extracted from all 15 of the non-clotted specimens had a RIN ≥7.0, only 9 of the 15 clotted specimens yielded RNA with a RIN ≥7.0. All 4 specimens with small clots yielded RNA with a RIN ≥7.0, but only 5 of the 11 specimens with large clots had RNA with RIN ≥7.0.

   Biospecimens

   • Bodily Fluid - Blood

   Preservative Types

   • Frozen

   Diagnoses:

   • Not specified

   Platform:

   Analyte	Technology Platform
   RNA	Automated electrophoresis/Bioanalyzer
   RNA	Spectrophotometry

   Pre-analytical Factors:

   Classification	Pre-analytical Factor	Value(s)
   Biospecimen Aliquots and Components	Blood and blood products	Clotted specimen Non clotted specimen

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