Kim YeJi, Lee Won-Nyoung, Yoo Hyun Jin, Baek Changyoon, Min Junhong
School of Integrative Engineering, Chung-Ang University, Heuksukdong, Dongjak-gu, Seoul, 06974 Republic of Korea.
Biochip J. 2017;11(4):255-261. doi: 10.1007/s13206-017-1401-y. Epub 2017 Dec 12.
Recently, a variety of methods, so called "direct 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, NaSO and GuSCN were selected as representative kosmotropic and chaotropic salts, respectively. MgCl 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 10 pfu/100 μL of whole blood by a very simple method.
最近，已经开发出了多种所谓的“直接缓冲液”方法，用于在现场无需任何设备的情况下利用血液中的核酸来检测感染性细菌和病毒。在此，我们首先研究了可能构成直接缓冲液的候选化学物质对全血中主要成分PCR抑制作用降低的单独和组合效果。分别选择长短链聚乙二醇、硫酸钠和硫氰酸胍作为典型的盐析盐和离液盐。选择氯化镁作为二价阳离子源，并使用氢氧化钠来控制血液的pH值。用 Triton X-100测试了常见非离子生物洗涤剂的效果，并选择十二烷基硫酸钠（SDS）作为阴离子洗涤剂。这些结果可为基于核酸的诊断领域中样品制备溶液的开发提供基础。结果，本研究中开发的直接缓冲液能够通过非常简单的方法检测浓度为10 pfu/100 μL全血的病毒。
