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  2. MALDI-TOF MS for rapid detection and differentiation between Tet(X)-producers and non-Tet(X)-producing tetracycline-resistant Gram-negative bacteria

MALDI-TOF MS for rapid detection and differentiation between Tet(X)-producers and non-Tet(X)-producing tetracycline-resistant Gram-negative bacteria

  • Virulence. 2022 Dec;13(1):77-88. doi: 10.1080/21505594.2021.2018768.
Zi-Jian Zheng 1 2 3 Ze-Hua Cui 1 2 3 Qiu-Yue Diao 1 2 3 Xin-Qing Ye 1 2 3 Zi-Xing Zhong 1 2 3 Tian Tang 1 2 3 Shuai-Bin Wu 1 2 3 Hui-Ling He 1 2 3 Xin-Lei Lian 1 2 3 Liang-Xing Fang 1 2 3 Xi-Ran Wang 1 2 3 Li-Jie Liang 1 2 3 Ya-Hong Liu 1 2 3 4 Xiao-Ping Liao 1 2 3 Jian Sun 1 2 3
Affiliations

Affiliations

  • 1 National Risk Assessment Laboratory for Antimicrobial Resistance of Animal Original Bacteria, South China Agricultural University, Guangzhou, China.
  • 2 Guangdong Laboratory for Lingnan Modern Agriculture, Guangzhou, China.
  • 3 Guangdong Provincial Key Laboratory of Veterinary Pharmaceutics Development and Safety Evaluation, South China Agricultural University, Guangzhou, China.
  • 4 Jiangsu Co-Innovation Center for the Prevention and Control of Important Animal Infectious Disease and Zoonosis, Yangzhou University, Yangzhou, China.
Abstract

The extensive use of Tetracycline antibiotics has led to the widespread presence of tetracycline-resistance genes in Gram-negative bacteria and this poses serious threats to human and animal health. In our previous study, we reported a method for rapid detection of Tet(X)-producers using MALDI-TOF MS. However, there have been multiple machineries involved in Tetracycline resistance including efflux pump, and ribosomal protection protein. Our previous demonstrated the limitation in probing the non-Tet(X)-producing tetracycline-resistant strains. In this regard, we further developed a MALDI-TOF MS method to detect and differentiate Tet(X)-producers and non-Tet(X)-producing tetracycline-resistant strains. Test strains were incubated with tigecycline and oxytetracycline in separate tubes for 3 h and then analyzed spectral peaks of tigecycline, oxytetracycline, and their metabolite. Strains were distinguished using MS ratio for [metabolite/(metabolite+ tigecycline or oxytetracycline)]. Four control strains and 319 test strains were analyzed and the sensitivity was 98.90% and specificity was 98.34%. This was consistent with the results obtained from LC-MS/MS analysis. Interestingly, we also found that the Reactive Oxygen Species (ROS) produced by tetracycline-susceptible strains were able to promote the degradation of oxytetracycline. Overall, the MALDITet(X)-plus test represents a rapid and reliable method to detect Tet(X)-producers, non-Tet(X)-producing tetracycline-resistant strains, and tetracycline-susceptible strains.

Keywords

MALDI-TOF MS; Rapid detection; Tet(X)-producers; non-Tet(X)-producers; phenotypic detection; tetracycline resistance.

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