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Updated: Jan 25, 2026

Development of a Quantitative Recombinase Polymerase Amplification Assay with an Internal Positive Control
Published on: March 30, 2015
[Establishment of recombinase polymerase amplification assay for drug resistance gene bla_(NDM) coding
Bowen Tu1, Ying Zhao1, Junhong Li1
1Inspection Center of Changzhou Disease Control and Prevention Center, Changzhou 213022, China.
Objective:
To establish a recombinase polymerase amplification(RPA) method for extensively resistant pathogen screening and rapid detection in the field for rapid amplification of the metallo-beta-lactamase gene bla_(NDM).
Methods:
Specific conservative sequence had been selected as target genes by sequence comparative analysis. The primers and probes for RPA assays were designed according to the principle of RPA amplification requirements. A descending gradient diluted template genes were used for RPA detection to determine the sensitivity. Reference templates of other resistant types of bacteria were used to analysis the specificity of amplification reaction systems. The amplification reaction systems were also conducted repeatedly for verifying the repeatability.
Results:
Three of the RPA reaction systems could effectively amplify the target genes, the sensitivities reach 2×10~2 copies. No one cross reaction existed with the other drug-resistant bacteria DNA. All the reactions can be completed between two to seven minutes.
Conclusion:
The RPA assays of the metallo-beta-lactamase gene bla_(NDM) are established, which may amply target genes fast and have a lower detection limit, and be potentially useful for in field pathogens detection.
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