title: A novel diagnostic platform based on multiplex ligase detection-PCR and microarray for simultaneous detection of swine viruses.
abstract: Simultaneous detection and identification of multiple pathogens is required in many diagnostic fields. In this study a novel method based on a multiplex ligase detection (LD)-polymerase chain reaction (PCR) and microarray (MLPM) is described to detect simultaneously several swine viruses involved in reproductive and/or respiratory problems. The multiplex diagnostic system was validated using standard plasmids, and clinical samples. Using this strategy as few as 10 copies of target plasmids were detected successfully. Each probe pair yielded specific positive signal only in its target site. In addition, when six target plasmids were present simultaneously sufficient robust signals were generated in their corresponding sites of six plasmid templates and no obvious signals were detected in non-target sites. Compared to real-time PCR, the MLPM showed specificities and sensitivities of 95.7-100% and 100% for 47 clinical samples tested, respectively. The results demonstrate that this novel assay is a specific, sensitive, and multiplex diagnostic method for detection of multiple pathogens and can also be adapted easily for diagnostic purposes.
authors.affiliation: College of Life Sciences, Zhejiang Sci-Tech University, 2nd Street, Hangzhou 310018, China. yonghoujiang@zstu.edu.cn
authors.forename: Yonghou
authors.initials: Y
authors.lastname: Jiang
authors.forename: Yao
authors.initials: Y
authors.lastname: Guo
authors.forename: Ping
authors.initials: P
authors.lastname: Wang
authors.forename: Qinfang
authors.initials: Q
authors.lastname: Dong
authors.forename: Tanja
authors.initials: T
authors.lastname: Opriessnig
authors.forename: Juhui
authors.initials: J
authors.lastname: Cheng
authors.forename: Hui
authors.initials: H
authors.lastname: Xu
authors.forename: Xianfeng
authors.initials: X
authors.lastname: Ding
authors.forename: Jiangfeng
authors.initials: J
authors.lastname: Guo
mesh_terms: D000818:Animals; D019411:Clinical Laboratory Techniques / Q000379:methods*; D005260:Female; D052776:Female Urogenital Diseases / Q000175:diagnosis / Q000662:veterinary / Q000821:virology; D008025:Ligases; D008297:Male; D052801:Male Urogenital Diseases / Q000175:diagnosis / Q000662:veterinary / Q000821:virology; D046228:Microarray Analysis / Q000379:methods*; D016133:Polymerase Chain Reaction / Q000379:methods*; D012141:Respiratory Tract Infections / Q000175:diagnosis / Q000662:veterinary / Q000821:virology; D012680:Sensitivity and Specificity; D013552:Swine / Q000821:virology; D013553:Swine Diseases / Q000175:diagnosis* / Q000821:virology*; D014730:Veterinary Medicine / Q000379:methods; D014773:Virology / Q000379:methods*; D014777:Virus Diseases / Q000175:diagnosis / Q000662:veterinary* / Q000821:virology
chemical_list: D008025:Ligases