Establishing rapid PCR/LAMP screening detection methods for NDM-1superbug

Objective:Establishing rapid PCR/LAMP screening detection methods for NDM-1(New Delhi Metallo-beta-Lactamase-1)superbug.Methods: Designing the primers of PCR/LAMP for superbug based on the sequences of NDM-1 from Genbank and optimizing the reaction conditions for PCR/LAMP.Forty one standard strains,one positive clone and two hundred and seventy five strains collected from different regions were tested for the specificity and sensitivity of PCR/LAMP methods.Results: These two methods show good specificity,and no strain was positvie except the positive clone.The detection limits of these two methods were different,with detection limits of 31 cfu/reaction and 123 cfu/reaction for LAMP and PCR,respectively.LAMP can be finished in one step in 1.5 to 2 hours from the DNA extraction to resμlts identification,however,it was time consuming for PCR and need about 3~4 hours for the whole process.Conclusion: The specificity and sensitivity of PCR/LAMP methods established for the detection of superbug NDM-1gene appeared to be pretty good,and these two methods can be acted as one of the emergency detection reserve methods and used in the monitoring survey of superbug now.