Institute of Nano Electronic Engineering, Universiti Malaysia Perlis (UniMAP), Kangar, Perlis, 01000, Malaysia.
Faculty of Chemical Engineering Technology, Universiti Malaysia Perlis (UniMAP), Arau, Perlis, 02600, Malaysia.
Biotechnol Appl Biochem. 2022 Aug;69(4):1696-1711. doi: 10.1002/bab.2239. Epub 2021 Aug 22.
An oligonucleotide DNA probe has been developed for the application in the DNA electrochemical biosensor for the early diagnosis of coronavirus disease (COVID-19). Here, the virus microRNA from the N-gene of severe acute respiratory syndrome-2 (SARS-CoV-2) was used for the first time as a specific target for detecting the virus and became a framework for developing the complementary DNA probe. The sequence analysis of the virus microRNA was carried out using bioinformatics tools including basic local alignment search tools, multiple sequence alignment from CLUSTLW, microRNA database (miRbase), microRNA target database, and gene analysis. Cross-validation of distinct strains of coronavirus and human microRNA sequences was completed to validate the percentage of identical and consent regions. The percent identity parameter from the bioinformatics tools revealed the virus microRNAs' sequence has a 100% match with the genome of SARS-CoV-2 compared with other coronavirus strains, hence improving the selectivity of the complementary DNA probe. The 30 mer with 53.0% GC content of complementary DNA probe 5' GCC TGA GTT GAG TCA GCA CTG CTC ATG GAT 3' was designed and could be used as a bioreceptor for the biosensor development in the clinical and environmental diagnosis of COVID-19.
已经开发出一种寡核苷酸 DNA 探针,用于冠状病毒病 (COVID-19) 的 DNA 电化学生物传感器的早期诊断。在这里,首次将严重急性呼吸系统综合征 2 型 (SARS-CoV-2) 的 N 基因中的病毒 microRNA 用作检测病毒的特定靶标,并成为开发互补 DNA 探针的框架。使用生物信息学工具(包括基本局部比对搜索工具、来自 CLUSTLW 的多序列比对、microRNA 数据库 (miRbase)、microRNA 靶标数据库和基因分析)对病毒 microRNA 的序列进行了分析。完成了不同冠状病毒株和人类 microRNA 序列的交叉验证,以验证相同和一致区域的百分比。来自生物信息学工具的同一性参数表明,与其他冠状病毒株相比,病毒 microRNAs 的序列与 SARS-CoV-2 的基因组具有 100%的匹配性,从而提高了互补 DNA 探针的选择性。设计了 30 个碱基、53.0%GC 含量的互补 DNA 探针 5' GCC TGA GTT GAG TCA GCA CTG CTC ATG GAT 3',可作为生物传感器的生物受体,用于 COVID-19 的临床和环境诊断。