title: Development of a real-time polymerase chain reaction assay for detection of Actinobacillus suis in porcine lung.
abstract: In the current study, the development and validation of a real-time polymerase chain reaction (PCR) assay using a TaqMan-labeled probe for the detection of Actinobacillus suis from porcine lung samples is described. This real-time PCR amplified a 110-bp region of the 23S ribosomal RNA gene from A. suis but not from other bacteria. First, the assay was validated with 183 bacterial strains representing different species of bacteria. Subsequently, 85 porcine lung specimens that were declared A. suis-positive and -negative by bacterial culture and identification were tested to assess whether it can be performed directly on tissue specimens. The bacterial culture results and real-time PCR results agreed across all the samples tested assigning 100% positive and negative predictive values to the PCR. Further, the detection limit of the assay was 380 colony-forming units (CFU) per ml or approximately 1 CFU per reaction. In conclusion, the TaqMan real-time PCR assay described herein is a highly specific, sensitive, and reproducible test, which can be used to detect A. suis DNA in porcine lung specimens, thus providing a timely diagnosis.
authors: Department of Veterinary and Biomedical Sciences, The Pennsylvania State University, University Park, PA 16802, USA. sxk91@psu.edu
authors: Subhashinie
authors: S
authors: Kariyawasam
authors: Erin
authors: E
authors: Strait
authors: Dianna
authors: D
authors: Jordan
authors: Jeremy
authors: J
authors: Kroll
mesh_terms: D000189:Actinobacillus Infections / Q000175:diagnosis / Q000382:microbiology / Q000662:veterinary*; D044132:Actinobacillus suis / Q000302:isolation & purification*; D000818:Animals; D008168:Lung / Q000382:microbiology*; D008171:Lung Diseases / Q000175:diagnosis / Q000382:microbiology / Q000662:veterinary*; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods / Q000662:veterinary*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013552:Swine; D013553:Swine Diseases / Q000175:diagnosis / Q000382:microbiology*