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Current Neurovascular Research


ISSN (Print): 1567-2026
ISSN (Online): 1875-5739

Research Article

Simple Electric Device to Isolate Nucleic Acids from Whole Blood Optimized for Point of Care Testing of Brain Damage

Author(s): Mi Jung Bae, Young Mi Lee, Ye Seul Choi, Eunmi Lee, Minh Tan Le, Thi Hong Duc Nguyen, Donghyeon Lee, Junghwan Cho, Hyung Soo Han*, Nora Jee-Young Park and Gun Oh Chong

Volume 19, Issue 3, 2022

Published on: 07 October, 2022

Page: [333 - 343] Pages: 11

DOI: 10.2174/1567202619666220903105805

open access plus


Background: Detection or monitoring of brain damage is a clinically crucial issue. Nucleic acids in the whole blood can be used as biomarkers for brain injury. Polymerase chain reaction (PCR) which is one of the most commonly used molecular diagnostic assays requires isolated nucleic acids to initiate amplification. Currently used nucleic acid isolation procedures are complicated and require laboratory equipments.

Objective: In this study, we tried to develop a simple and convenient method to isolate nucleic acids from the whole blood sample using a tiny battery-powered electric device. The quality of the isolated nucleic acids should be suitable for PCR assay without extra preparation.

Methods: A plastic device with separation chamber was designed and printed with a 3D printer. Two platinum electrodes were placed on both sides and a battery was used to supply the electricity. To choose the optimal nucleic acid isolation condition, diverse lysis buffers and separation buffers were evaluated, and the duration and voltage of the electricity were tested. Western blot analysis and PCR assay were used to determine the quality of the separated nucleic acids.

Results: 2ul of whole blood was applied to the cathode side of the separation chamber containing 78 ul of normal saline. When the electricity at 5 V was applied for 5 min, nucleic acids were separated from segment 1 to 3 of the separation chamber. The concentration of nucleic acids peaked around 7~8 mm from cathode side. PCR assay using the separation buffer as the template was performed successfully both in conventional and realtime PCR methods. The hemoglobin in the whole blood did not show the inhibitory effect in our separation system and it may be due to structural modification of hemoglobin during electric separation.

Conclusion: Our simple electric device can separate nucleic acids from the whole blood sample by applying electricity at 5 V for 5 min. The separation buffer solution taken from the device can be used for PCR assay successfully.

Keywords: Point of care test, whole blood, nucleic acid, electric device, brain damage, molecular diagnosis.

Katan M, Luft A. Global burden of stroke. Semin Neurol 2018; 38(2): 208-11.
Harpaz D, Eltzov E, Seet RCS, Marks RS, Tok AIY. Point-of-care-testing in acute stroke management: An unmet need ripe for technological harvest. Biosensors 2017; 7(4): 30.
[] [PMID: 28771209]
Mondello S, Sorinola A, Czeiter E, et al. Blood-based protein biomarkers for the management of traumatic brain injuries in adults presenting to emergency departments with mild brain injury: A living systematic review and meta-analysis. J Neurotrauma 2021; 38(8): 1086-106.
[] [PMID: 29020853]
Biberthaler P, Musaelyan K, Krieg S, et al. Evaluation of acute glial fibrillary acidic protein and ubiquitin C-Terminal Hydrolase-L1 plasma levels in traumatic brain injury patients with and without intracranial lesions. Neurotrauma Reports 2021; 2(1): 617-25.
[] [PMID: 35018363]
Davis CK, Vemuganti R. DNA damage and repair following traumatic brain injury. Neurobiol Dis 2021; 147105143.
[] [PMID: 33127471]
Campello Yurgel V, Ikuta N, Da Rocha BA, et al. Role of plasma DNA as a predictive marker of fatal outcome following severe head injury in males. J Neurotrauma 2007; 24(7): 1172-81.
[] [PMID: 17610356]
Rao A, Berg B, Quezada T, et al. Diagnosis and antibiotic treatment of group a streptococcal pharyngitis in children in a primary care setting: Impact of point-of-care polymerase chain reaction. BMC Pediatr 2019; 19(1): 24.
[] [PMID: 30651115]
Liu C, Xiang X, Han S, et al. Blood-based liquid biopsy: Insights into early detection and clinical management of lung cancer. Cancer Lett 2022; 524: 91-102.
[] [PMID: 34656690]
Trujillo B, Wu A, Wetterskog D, Attard G. Blood-based liquid biopsies for prostate cancer: Clinical opportunities and challenges. Br J Cancer 2022. [Epub Ahead of Print]
[] [PMID: 35715640]
Lone SN, Nisar S, Masoodi T, et al. Liquid biopsy: A step closer to transform diagnosis, prognosis and future of cancer treatments. Mol Cancer 2022; 21(1): 79.
[] [PMID: 35303879]
Wijerathne H, Witek MA, Baird AE, Soper SA. Liquid biopsy markers for stroke diagnosis. Expert Rev Mol Diagn 2020; 20(8): 771-88.
[] [PMID: 32500751]
Marcatti M, Saada J, Okereke I, et al. Quantification of circulating cell free mitochondrial dna in extracellular vesicles with PicoGreen™ in liquid biopsies: Fast assessment of disease/trauma severity. Cells 2021; 10(4): 819.
[] [PMID: 33917426]
Sullivan BP, Bender AT, Ngyuen DN, Zhang JY, Posner JD. Nucleic acid sample preparation from whole blood in a paper microfluidic device using isotachophoresis. J Chromatogr B Analyt Technol Biomed Life Sci 2021; 1163122494.
[] [PMID: 33401049]
Lee K, Kang JH, Kim HM, et al. Direct electrophoretic microRNA preparation from clinical samples using nanofilter membrane. Nano Converg 2020; 7(1): 1.
[] [PMID: 31930443]
Marshall L, Dyer R. Purification of FFPE samples using isotachophoresis to improve the yield and quality of amplifiable DNA. J Biomol Tech: JBT 2020; 31 (Suppl.): S11.
De Oliveira CB, Franco OL. Cryptic host defense peptides: Multifaceted activity and prospects for medicinal chemistry. Curr Top Med Chem 2020; 20(14): 1274-90.
[] [PMID: 32209042]
Sidstedt M, Hedman J, Romsos EL, et al. Inhibition mechanisms of hemoglobin, immunoglobulin G, and whole blood in digital and real-time PCR. Anal Bioanal Chem 2018; 410(10): 2569-83.
[] [PMID: 29504082]

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