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

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

Volume: 19 Issue: 3

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

Affiliation:

• Department of Physiology, School of Medicine, Kyungpook National University, Daegu 41944, Korea;
• Department of Biomedical Science, Graduate School, Kyungpook National University, Daegu 41944, Korea
• Clinical Omics Institute, Kyungpook National University, Daegu 41405, Korea
• BK21 Four Program, School of Medicine, Kyungpook National University, Daegu 41944, Korea

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

Abstract:

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.

Cite this article as:

Bae Jung Mi, Lee Mi Young, Choi Seul Ye, Lee Eunmi, Le Tan Minh, Duc Nguyen Thi Hong, Lee Donghyeon, Cho Junghwan, Han Soo Hyung*, Park Jee-Young Nora and Chong Oh Gun, Simple Electric Device to Isolate Nucleic Acids from Whole Blood Optimized for Point of Care Testing of Brain Damage, Current Neurovascular Research 2022; 19 (3) . https://dx.doi.org/10.2174/1567202619666220903105805