Direct buffer composition of blood pre-process for nucleic acid based diagnostics

核酸诊断血液预处理中直接缓冲液的组成

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Abstract

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, Na(2)SO(4) and GuSCN were selected as representative kosmotropic and chaotropic salts, respectively. MgCl(2) 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(2) pfu/100 μL of whole blood by a very simple method.

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