903 Protein Saver cards: the best alternative for dried blood spot storage at room temperature for HCV RNA.
Author(s): Arca-Lafuente S, Casanueva-Benítez C, Crespo-Bermejo C, Lara-Aguilar V, Martín-Carbonero L, de Los Santos I, Madrid R, Briz V
Publication: Sci Rep, 2022, Vol. 12, Page 10124
PubMed ID: 35710721 PubMed Review Paper? No
Purpose of Paper
The purpose of this paper was to compare the stability of hepatitis C virus (HCV) RNA in dried blood and plasma specimens that were stored at room temperature or at -20°C on two different types of cards (903 protein saver cards and FTA classic cards) for up to 390 days.
Conclusion of Paper
The number of specimens with detectable HCVa declined with longer storage on both types of cards. Stability of HCV RNA was slightly better for when plasma or blood was stored on Protein Saver Cards than on FTA Classic Cards. HCV RNA was more stable when specimens were stored on FTA Classic Cards at -20°C rather than at room temperature, but specimens on 903 Protein Saver Cards had slightly more stable HCV RNA when stored at room temperature than at -20°C. HCV was detectable in all blood and plasma specimens on 903 Protein Saver Cards stored for 255 days at room temperature, but the number of specimens with detectable HCV declined with longer storage. Sequencing of amplified HCV from blood on 903 Protein Saver Cards stored at room temperature for 255 days showed 97% sequence identity to fresh plasma specimens, but declined to 87% identity when stored for 285 days.
Studies
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Study Purpose
The purpose of this study was to compare the stability of HCV RNA in dried blood and plasma specimens that were stored on 903 protein saver cards and FTA classic cards for up to 390 days at room temperature or at -20°C. EDTA blood was collected from six patients with HCV. Plasma was obtained from 4 of the 5 mL of blood collected while the remaining 1 mL was directly applied to the cards directly (no other details of plasma processing were provided). Viral load in plasma was quantified using the COBAS TaqMan HCV Test. Blood and plasma were applied to FTA Classic Cards (200 µL) and 903 Protein Saver Cards (25 µL) and allowed to dry at room temperature for 2 h, after which they were stored at room temperature or at -20°C. At each timepoint a 3 mm punch was cut from each card, and RNA was purified using the Whatman Purification Reagent, and HCV was detected by RT-PCR. To evaluate HCV integrity, amplified nested PCR products were extracted from agarose gels, purified using the Wizard SV Gel and PCR Clean-Up System and Sanger sequenced.
Summary of Findings:
The number of specimens with detectable HCV declined with longer storage on both types of cards. Stability of HCV RNA was slightly better when blood or plasma were stored on Protein Saver Cards rather than FTA Classic Cards. HCV RNA stability in blood and plasma was also more stable when FTA Classic Cards were stored at -20°C rather than room temperature, or when903 Protein Saver Cards cards were stored at room temperature compared to -20°C. HCV RNA was detectable after 285 days in 9 of 11 specimens stored on FTA Classic Cards at -20°C but only detectable in 6 of 11 specimens stored at room temperature. After storage of FTA Classic Cards for 360 days, HCV RNA was detected in 2 of 6 blood specimens, regardless of storage temperature and in 2 and 0 of the 5 plasma specimens stored at -20°C and room temperature, respectively. After 255 days of storage at room temperature on 903 Protein Saver Cards, HCV was detectable in all blood and plasma specimens, and after 285 days at room temperature was detectable in 5 of 6 blood and 4 of 5 plasma specimens. After 360 days at room temperature only 4 of 11 (blood and plasma combined) specimens on 903 Protein Saver Cards had detectable HCV RNA. Sequencing of amplified HCV from blood stored on 903 Protein Saver Cards at room temperature for 255 days displayed a 97% sequence identity to that of fresh plasma specimens, but when stored for 285 days this declined to 87% identity and sequencing was not possible from blood specimens stored on 903 Protein Saver Cards for 360 days.
Biospecimens
Preservative Types
- Other Preservative
- Frozen
Diagnoses:
- Hepatitis
Platform:
Analyte Technology Platform RNA DNA sequencing RNA RT-PCR Pre-analytical Factors:
Classification Pre-analytical Factor Value(s) Biospecimen Aliquots and Components Blood and blood products Whole blood
Plasma
Storage Storage conditions 903 Protein Saver Cards
FTA classic cards
As dried blood
As dried plasma
Storage Storage temperature Room temperature
-20°C
Storage Storage duration 0 days
15 days
60 days
120 days
165 days
195 days
255 days
285 days
360 days
390 days