Open AccessSimultaneous Detection of Actinobacillus pleuropneumoniae and Haemophilus parasuis in Pig by Duplex Droplet Digital PCR
Author(s) -
Wen Wen,
Shi Lei,
Chen Xun
Publication year - 2020
Publication title -
efood
Language(s) - English
Resource type - Journals
ISSN - 2666-3066
DOI - 10.2991/efood.k.200918.001
Subject(s) - actinobacillus pleuropneumoniae , haemophilus , digital polymerase chain reaction , microbiology and biotechnology , biology , polymerase chain reaction , virology , bacteria , serotype , genetics , gene
Actinobacillus pleuropneumoniae ( A. pleuropneumoniae ) and Haemophilus parasuis ( H. parasuis ) are important pathogens of swine, which cause great economic losses in the swine industry and bring great safety hazards to food safety. In this study, we established a duplex Droplet Digital Polymerase Chain Reaction (ddPCR) assay for sensitive detection of A. pleuropneumoniae and H. parasuis simultaneously in pork. The limit of detection of plasmid DNA could reach 2 copies/μL for both A. pleuropneumoniae and H. parasuis ; which were approximately 50‐ and 20‐fold greater sensitivity than Quantitative real‐time PCR (qPCR). Both qPCR and ddPCR showed high linearity and positive correlation with standards. The detection results of clinical suspected samples for A. pleuropneumoniae and H. parasuis indicated the positive detection rate of ddPCR (10.7% and 12.5%) was higher than that of qPCR (7.1% and 8.9%). Therefore, the duplex ddPCR assay could be used as an effective quantitative detection to diagnose A. pleuropneumoniae and H. parasuis simultaneously, which may lay a foundation for the development of substitute rapid diagnostic technique.