Selective detection of viable seed-borne Acidovorax citrulli by real-time PCR with propidium monoazide.

ABSTRACT: In recent years, use of the DNA-intercalating dye propidium monoazide (PMA) in real-time PCR has been reported as a novel method to detect viable bacteria in different types of samples, such as food, environmental, and microbiological samples. In this study, viable cells of Acidovorax citrulli, the causal agent of bacterial seedling blight and fruit blotch, were selectively detected and differentiated from dead cells by real-time fluorescent polymerase chain reaction amplification after the bacterial solution was treated with the DNA-binding dye PMA. The primers and TaqMan probe were based on the A. citrulli genome (Aave_1909, Gene ID: 4669443) and were highly specific for A. citrulli. The detection threshold of this assay was 10³ colony-forming units per mL (CFU/mL) in pure cel

SUBMITTER: Tian Q 

PROVIDER: S-EPMC5064318 | biostudies-literature | 2016 Oct

REPOSITORIES: biostudies-literature