Shi-Jun Zhang: Key Laboratory of Zoonosis Research, Ministry of Education, Institute of Zoonosis, College of Veterinary Medicine, College of Animal Sciences, Jilin University, Xi An Da Lu 5333, Changchun 130062, China.
Lu-Lu Wang: Key Laboratory of Zoonosis Research, Ministry of Education, Institute of Zoonosis, College of Veterinary Medicine, College of Animal Sciences, Jilin University, Xi An Da Lu 5333, Changchun 130062, China.
Shi-Ying Lu: Key Laboratory of Zoonosis Research, Ministry of Education, Institute of Zoonosis, College of Veterinary Medicine, College of Animal Sciences, Jilin University, Xi An Da Lu 5333, Changchun 130062, China.
Pan Hu: Key Laboratory of Zoonosis Research, Ministry of Education, Institute of Zoonosis, College of Veterinary Medicine, College of Animal Sciences, Jilin University, Xi An Da Lu 5333, Changchun 130062, China.
Yan-Song Li: Key Laboratory of Zoonosis Research, Ministry of Education, Institute of Zoonosis, College of Veterinary Medicine, College of Animal Sciences, Jilin University, Xi An Da Lu 5333, Changchun 130062, China.
Ying Zhang: Key Laboratory of Zoonosis Research, Ministry of Education, Institute of Zoonosis, College of Veterinary Medicine, College of Animal Sciences, Jilin University, Xi An Da Lu 5333, Changchun 130062, China.
Heng-Zhen Chang: Key Laboratory of Zoonosis Research, Ministry of Education, Institute of Zoonosis, College of Veterinary Medicine, College of Animal Sciences, Jilin University, Xi An Da Lu 5333, Changchun 130062, China.
Fei-Fei Zhai: Key Laboratory of Zoonosis Research, Ministry of Education, Institute of Zoonosis, College of Veterinary Medicine, College of Animal Sciences, Jilin University, Xi An Da Lu 5333, Changchun 130062, China.
Zeng-Shan Liu: Key Laboratory of Zoonosis Research, Ministry of Education, Institute of Zoonosis, College of Veterinary Medicine, College of Animal Sciences, Jilin University, Xi An Da Lu 5333, Changchun 130062, China.
Zhao-Hui Li: Key Laboratory of Zoonosis Research, Ministry of Education, Institute of Zoonosis, College of Veterinary Medicine, College of Animal Sciences, Jilin University, Xi An Da Lu 5333, Changchun 130062, China.
Hong-Lin Ren: Key Laboratory of Zoonosis Research, Ministry of Education, Institute of Zoonosis, College of Veterinary Medicine, College of Animal Sciences, Jilin University, Xi An Da Lu 5333, Changchun 130062, China.
INTRODUCTION: The plate counting method widely used at present to discern viable from non-viable in the host or cell is time-consuming and laborious. Therefore, it is necessary to establish a rapid, simple method for detecting and counting viable organisms. MATERIAL AND METHODS: Using propidium monoazide (PMA) to inhibit amplification of DNA from dead , a novel, rapid PMA-quantitative PCR (PMA-qPCR) detection method for counting viable was established. The standard recombinant plasmid with the target gene fragment inserted was constructed for drawing a standard curve. The reaction conditions were optimised, and the sensitivity, specificity, and repeatability were analysed. RESULTS: The optimal exposure time and working concentration of PMA were 10 min and 15 μg/mL, respectively. The correlation coefficient (R) of the standard curve was 0.999. The sensitivity of the method was 10 CFU/mL, moreover, its specificity and repeatability also met the requirements. The concentration of measured by the PMA-qPCR did not differ significantly from that measured by the plate counting method, and the concentrations of viable bacteria in infected cells determined by the two methods were of the same order of magnitude. CONCLUSION: In this study, a rapid and simple PMA-qPCR counting method for viable was established, which will facilitate related research.
