Establishment of an A/T-Rich Specifically MGB Probe digital droplet PCR Assays Based on SNP for Brucella wild strains and vaccine strains.

Autor: Li W; Medical College, Inner Mongolia Minzu University, Tongliao 028000, China., Zhang S; Medical College, Inner Mongolia Minzu University, Tongliao 028000, China., Dang S; Keerqin District First People's Hospital, Tongliao 028000, China., Gao L; Tongliao Infectious Disease Hospital, Tongliao 028000, China., Li G; Tongliao Infectious Disease Hospital, Tongliao 028000, China., Cheng D; Beidahuang Industry Group General Hospital, Harbin 150000, China., Jiang L; College of Chemistry, Fuzhou University, Fuzhou 350000, China., Huang T; College of Public Health, Inner Mongolia Minzu University, Tongliao 028000, China; Brucellosis Prevention and Treatment Engineering Research Center of Inner Mongolia Autonomous Region, Tongliao 028000, China; Key Laboratory of Zoonose Prevention and Control at Universities of Inner Mongolia Autonomous Region, Tongliao 028000, China., Zhai J; Medical College, Inner Mongolia Minzu University, Tongliao 028000, China; Brucellosis Prevention and Treatment Engineering Research Center of Inner Mongolia Autonomous Region, Tongliao 028000, China; Key Laboratory of Zoonose Prevention and Control at Universities of Inner Mongolia Autonomous Region, Tongliao 028000, China. Electronic address: jbzhai@imun.edu.cn.
Jazyk: angličtina
Zdroj: Diagnostic microbiology and infectious disease [Diagn Microbiol Infect Dis] 2024 Sep; Vol. 110 (1), pp. 116432. Date of Electronic Publication: 2024 Jul 17.
DOI: 10.1016/j.diagmicrobio.2024.116432
Abstrakt: In recent years, immunization with the S2 live-attenuated vaccine has been recognized as the most economical and effective strategy for preventing brucellosis in Inner Mongolia, China. However, there are still challenges related to vaccine toxicity and the inability to distinguish between vaccine immunization and natural infection. Therefore, in this study, we developed a digital droplet polymerase chain reaction (ddPCR) assay based on single-nucleotide polymorphism (SNP) loci to identify wild Brucella strains and S2 vaccine strains. The assay demonstrated excellent linearity (R 2 > 0.99) with a lower detection limit of 10 copies/µL for both wild and vaccine strains. Additionally, the ddPCR assay outperformed the real-time fluorescent quantitative PCR (qPCR) assay in screening 50 clinical samples. We have established an effective and highly sensitive ddPCR assay for Brucella, providing an efficient method for detecting and differentiating wild strains of Brucella from the S2 vaccine strain.
Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.
(Copyright © 2024 The Author(s). Published by Elsevier Inc. All rights reserved.)
Databáze: MEDLINE
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