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Info. Vol.11 - No.4 (2017.12.20)
Title Direct Buffer Composition of Blood Pre-process for Nucleic Acid Based Diagnostics
Authors YeJi Kim1, Won-Nyoung Lee1, Hyun Jin Yoo1, Changyoon Baek1 & Junhong Min1,*
Institutions 1School of Integrative Engineering, Chung-Ang University, Heuksukdong, Dongjak-gu, Seoul 06974, Republic of Korea
*Correspondence and requests for materials should be addressed to J. Min (junmin@cau.ac.kr)
Abstract Recently, a variety of methods, so called ?쐂irect buffer?? have been developed to utilize nucleic acid in the blood for the measurement of infectious bacteria and virus without any equipment in the field. In here, we first investigated the individual and combinatory effects of candidate chemicals which might be composed of the direct buffer on the PCR inhibition reduction of main compositions in whole blood. The long and short PEGs, Na2SO4 and GuSCN were selected as representative kosmotropic and chaotropic salts, respectively. MgCl2 were chosen as divalent cation source and NaOH was used to control blood pH. The effect of common non-ionic biological detergent was tested with Triton X-100 and SDS (Sodium Dodecyl sulfate) was chosen as anionic detergent. These results could provide a foundation for the development of sample preparation solution in nucleic acid based diagnostic field. As a result, the direct buffer developed in this study was able to detect viruses with a concentration of 102 pfu/100 關L of whole blood by a very simple method.
Keyword Whole blood, Adenovirus, Direct buffer, Nucleic acid, Sample preparation
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