title: Development of colloidal gold-based immunochromatographic assay for rapid detection of Mycoplasma suis in porcine plasma.
abstract: A one-step immunochromatographic assay using gold nanoparticles coated with polyclonal antibody (pAb) against Mycoplasma suis (M. suis) was developed in this study for the detection of M. suis in porcine plasma. The colloidal gold was prepared by the reduction of gold salt with sodium citrate coupled with pAb against M. suis. The pAb was produced by immunizing the BALB/c mice with recombinant MSG1 (rMSG1) protein from M. suis expressed in Escherichia coli. The optimal concentrations of the capture antibody and the coating antibody were 12 μg/ml and 1.5 mg/ml, respectively, and that of the blocking buffer was 1% bovine serum albumin. The lower detection limit of the immunochromatographic assay test was 100 ng/ml with visual detection under optimal conditions of analysis. Classical swine fever virus, porcine reproductive and respiratory syndrome virus, swine pneumonia mycoplasma, swine toxoplasma, and porcine parvovirus were used to evaluate the specificity of the immunochromatographic strips. No cross-reaction of the antibodies with other related swine pathogens was observed. This qualitative test based on the visual evaluation of the results did not require any equipment. The assay time for M. suis detection was less than 10 min, suitable for rapid detection at the grassroots level. The one-step colloidal gold immunochromatographic strips that we developed had high specificity and sensitivity. Therefore, this method would be feasible, convenient, rapid, and effective for detecting M. suis in porcine plasma.
authors.affiliation: College of Veterinary Medicine, Research Center for Animal Disease Control Engineering Shandong Province, Shandong Agricultural University, Tai'an 271018, China.
authors.forename: Kai
authors.initials: K
authors.lastname: Meng
authors.affiliation: College of Veterinary Medicine, Research Center for Animal Disease Control Engineering Shandong Province, Shandong Agricultural University, Tai'an 271018, China.
authors.forename: Wenjing
authors.initials: W
authors.lastname: Sun
authors.affiliation: College of Veterinary Medicine, Research Center for Animal Disease Control Engineering Shandong Province, Shandong Agricultural University, Tai'an 271018, China.
authors.forename: Peng
authors.initials: P
authors.lastname: Zhao
authors.affiliation: College of Veterinary Medicine, Research Center for Animal Disease Control Engineering Shandong Province, Shandong Agricultural University, Tai'an 271018, China.
authors.forename: Limei
authors.initials: L
authors.lastname: Zhang
authors.affiliation: College of Veterinary Medicine, Research Center for Animal Disease Control Engineering Shandong Province, Shandong Agricultural University, Tai'an 271018, China.
authors.forename: Dongjie
authors.initials: D
authors.lastname: Cai
authors.affiliation: College of Veterinary Medicine, Research Center for Animal Disease Control Engineering Shandong Province, Shandong Agricultural University, Tai'an 271018, China.
authors.forename: Ziqiang
authors.initials: Z
authors.lastname: Cheng
authors.affiliation: College of Veterinary Medicine, Research Center for Animal Disease Control Engineering Shandong Province, Shandong Agricultural University, Tai'an 271018, China.
authors.forename: Huijun
authors.initials: H
authors.lastname: Guo
authors.affiliation: College of Veterinary Medicine, Research Center for Animal Disease Control Engineering Shandong Province, Shandong Agricultural University, Tai'an 271018, China. Electronic address: Liujz@sdau.edu.cn.
authors.forename: Jianzhu
authors.initials: J
authors.lastname: Liu
authors.affiliation: College of Veterinary Medicine, Research Center for Animal Disease Control Engineering Shandong Province, Shandong Agricultural University, Tai'an 271018, China.
authors.forename: Dubao
authors.initials: D
authors.lastname: Yang
authors.affiliation: College of Veterinary Medicine, Research Center for Animal Disease Control Engineering Shandong Province, Shandong Agricultural University, Tai'an 271018, China.
authors.forename: Shujing
authors.initials: S
authors.lastname: Wang
authors.affiliation: College of Veterinary Medicine, Research Center for Animal Disease Control Engineering Shandong Province, Shandong Agricultural University, Tai'an 271018, China.
authors.forename: Tongjie
authors.initials: T
authors.lastname: Chai
mesh_terms: D000818:Animals; D000906:Antibodies / Q000276:immunology*; D000942:Antigens, Bacterial / Q000097:blood* / Q000276:immunology; D015374:Biosensing Techniques / Q000295:instrumentation; D002846:Chromatography, Affinity / Q000295:instrumentation*; D003102:Colloids / Q000737:chemistry; D004867:Equipment Design; D019544:Equipment Failure Analysis; D006046:Gold / Q000737:chemistry*; D053768:Metal Nanoparticles / Q000737:chemistry; D051379:Mice; D008807:Mice, Inbred BALB C; D009174:Mycoplasma / Q000276:immunology* / Q000302:isolation & purification*; D045729:Pneumonia of Swine, Mycoplasmal / Q000097:blood* / Q000175:diagnosis / Q000276:immunology; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013552:Swine
chemical_list: D000906:Antibodies; D000942:Antigens, Bacterial; D003102:Colloids; D006046:Gold
keywords: Colloidal gold; Immunochromatographic assay; Mycoplasma suis; Polyclonal antibody; Rapid detection