Molecular Detection of the mcr Genes by Multiplex PCR

Jiayang Liu1 ,* Zhiyuan Zhang1 ,* Yuyang Feng,1 Huimin Hu,1 Yu Yu,1 Lihao Qiu,1 Hongtao Liu,1 Zhimin Guo,2 Jing Huang,2 Chongtao Du,1 Jiazhang Qiu1 1Key Laboratory of Zoonosis, Ministry of Education, College of Veterinary Medicine, Jilin University, Changchun 130062, People’s Republic of China; 2Department of Clinical Laboratory, The First Hospital of Jilin University, Changchun 130021, People’s Republic of China*These authors contributed equally to this workCorrespondence: Jiazhang Qiu; Chongtao DuKey Laboratory of Zoonosis, Ministry of Education, College of Veterinary Medicine, Jilin University, Changchun 130062, People’s Republic of ChinaEmail qiujz@jlu.edu.cn; duct@jlu.edu.cnBackground: The emergence and prevalence of plasmid-mediated colistin-resistant bacterial strains in recent years have raised great concerns in clinical medicine. It is urgently needed to develop a cheaper, faster, simpler, sensitive, and specific molecular detection method to identify and monitor the dissemination of the transferable resistant determinants.Methods and Results: Herein, eight pairs of primers were designed to set up a multiplex PCR method for the rapid and efficient determination of reported mcr genes. This assay can give results within 85 min (35 min for amplification and 50 min for electrophoresis). We validated the feasibility of this assay by testing the presence of mcr genes in 60 colistin-resistant isolates.Conclusion: Our multiplex PCR technique exhibits remarkable advantages in the light of clear identification, efficiency of amplification, as well as the time consuming for detection, and thus could be useful for the surveillance and epidemiological research of plasmid-mediated colistin resistance, particularly for the under-resourced laboratories.Keywords: multiplex PCR, colistin resistance, mcr genes