title: Rapid and sensitive detection of Mycoplasma synoviae by an insulated isothermal polymerase chain reaction-based assay on a field-deployable device.
abstract: Mycoplasma synoviae (MS), causing respiratory diseases, arthritis, and eggshell apex abnormalities in avian species, is an important pathogen in the poultry industry. Implementation of a biosecurity plan is important in MS infection management. Working on a field-deployable POCKIT™ device, an insulated isothermal polymerase chain reaction (iiPCR) assay has a potential for timely MS detection on the farm. The MS iiPCR assay had limit of detection 95% of about 9 genome equivalents by testing serial dilutions of a standard DNA. The detection endpoint of the assay for detection of MS genomic DNA was comparable to a reference real-time PCR. The assay did not crossreact with other important avian pathogens, including avian reovirus, Mycoplasma gallisepticum, Staphylococcus aureus, Escherichia coli, Pasteurella multocida, and Salmonella Pullorum. When 92 synovial fluid and respiratory tract swab samples collected from chickens, turkeys, and geese suspected of MS infection were tested, the clinical performance of the MS iiPCR had 97.8% agreement (Cohen's kappa value, 0.95) with that of the reference real-time PCR. In conclusion, the MS iiPCR/POCKIT™ system, working with field-deployable manual or automatic nucleic acid extraction methods, has potential to serve as a rapid and sensitive on-site tool to facilitate timely detection of MS.
authors.affiliation: Yunlin-Chiayi-Tainan of Animal Disease Diagnostic Center, Department of Veterinary Medicine, National Chiayi University, Chiayi, Taiwan.
authors.forename: Hung-Chih
authors.initials: HC
authors.lastname: Kuo
authors.affiliation: Yunlin-Chiayi-Tainan of Animal Disease Diagnostic Center, Department of Veterinary Medicine, National Chiayi University, Chiayi, Taiwan.
authors.forename: Dan-Yuan
authors.initials: DY
authors.lastname: Lo
authors.affiliation: Yunlin-Chiayi-Tainan of Animal Disease Diagnostic Center, Department of Veterinary Medicine, National Chiayi University, Chiayi, Taiwan.
authors.forename: Chiou-Lin
authors.initials: CL
authors.lastname: Chen
authors.affiliation: GeneReach Biotech, Taichung, Taiwan.
authors.forename: Yun-Long
authors.initials: YL
authors.lastname: Tsai
authors.affiliation: GeneReach Biotech, Taichung, Taiwan.
authors.forename: Jia-Fong
authors.initials: JF
authors.lastname: Ping
authors.affiliation: GeneReach Biotech, Taichung, Taiwan.
authors.forename: Chien-Hsien
authors.initials: CH
authors.lastname: Lee
authors.affiliation: GeneReach Biotech, Taichung, Taiwan peiyu329@genereachbiotech.com.
authors.forename: Pei-Yu Alison
authors.initials: PA
authors.lastname: Lee
authors.affiliation: GeneReach Biotech, Taichung, Taiwan.
authors.forename: Hsiao-Fen Grace
authors.initials: HG
authors.lastname: Chang
mesh_terms: D000818:Animals; D001426:Bacterial Proteins / Q000302:isolation & purification*; D002645:Chickens; D037102:Lectins / Q000302:isolation & purification*; D009175:Mycoplasma Infections / Q000175:diagnosis / Q000382:microbiology / Q000662:veterinary*; D045805:Mycoplasma synoviae / Q000302:isolation & purification*; D011201:Poultry Diseases / Q000175:diagnosis* / Q000382:microbiology; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods / Q000662:veterinary*; D012137:Respiratory System / Q000382:microbiology; D012680:Sensitivity and Specificity; D013582:Synovial Fluid / Q000382:microbiology
chemical_list: D001426:Bacterial Proteins; D037102:Lectins; C406455:VlhA protein, Mycoplasma synoviae
keywords: Mycoplasma synoviae; insulated isothermal PCR; point-of-need detection