Expression of recombinant Bst Large Fragment polymerase and its application in isothermal amplification for detecting Neisseria meningitidis

  • Đào Thị Huyền Bệnh viện Trung ương Quân đội 108
  • Đào Thanh Quyên Bệnh viện Trung ương Quân đội 108
  • Trần Thị Thanh Huyền Bệnh viện Trung ương Quân đội 108
  • Trần Thị Thu Hiền Bệnh viện Trung ương Quân đội 108
  • Ngô Tất Trung Bệnh viện Trung ương Quân đội 108
  • Lê Hữu Song Bệnh viện Trung ương Quân đội 108

Main Article Content

Keywords

Bst Large Fragment, N. meningitidis, Loop-mediated isothermal amplification

Abstract

Objective: Expression of Bst Large Fragment enzyme (BstLF) and its application in isothermal amplification for detecting N. meningitidis. Subject and method: E. coli BL21(DE3) pLysS cells were transformed with pET15b_BstLF_6XHis encoding plasmid was then induced with the specific inducer (IPTG); Using the affinity chromatography method. Its function was evaluated and compared with the commercial enzyme in the LAMP reaction for identifying N. meningitidis. Result: The best condition for expression of high-quality recombinant Bst Large Fragment enzyme in 37oC, 1mM IPTG in four hours. The reaction using an in-house enzyme for detecting N. meningitidis was established in 50 minutes. The sensitivity achieved was 102 copies/reaction (confidence is 95%). Conclusion: Expression of BstLF enzyme and its application in LAMP assay for detecting N. meningitidis with a detection limit of 102 copies/reaction, equivalent to commercial Bst 3.0 DNA Polymerase enzyme.

Article Details

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Published
Aug 8, 2023
DOI: https://doi.org/10.52389/ydls.v18i5.1882
Issue
Volume 18-No 5/2023
Section
Articles
How to Cite
Huyền Đào T., Quyên Đào T., HuyềnT. T. T., HiềnT. T. T., TrungN. T., & SongL. H. (2023). Expression of recombinant Bst Large Fragment polymerase and its application in isothermal amplification for detecting Neisseria meningitidis. Tạp Chí Y Dược lâm sàng 108, 18(5). https://doi.org/10.52389/ydls.v18i5.1882