title: Development of an Indirect Dot-PPA-ELISA using glutamate dehydrogenase as a diagnostic antigen for the rapid and specific detection of Streptococcus suis and its application to clinical specimens.
abstract: Streptococcus suis is an important zoonotic pathogen causing infections in pigs and humans. Bacterial surface-related proteins are often explored as potential vaccine candidates and diagnostic antigens. In the present study, glutamate dehydrogenase, a highly conserved immunogenic extracellular protein, was used to establish a dot horseradish peroxidase enzyme-linked staphylococcal protein A immunosorbent assay (Dot-PPA-ELISA) for diagnosis of S. suis infection. The antigen-antibody reaction was optimised through checkerboard titration involving serial dilutions, followed by selective blocking tests and evaluations of cross-reaction, repeatability, and stability. Comparative analysis by using a conventional plate ELISA kit showed that the specificity and sensitivity of the Dot-PPA-ELISA were 97.5 and 96.6%, respectively. Furthermore, dynamic changes in the levels of antibody in rabbits immunised with a propolis inactivated vaccine were monitored by Dot-PPA-ELISA. A total seroprevalence of 73.1% in 305 pig serum samples indicated the method's applicability to detect S. suis infection. Cumulatively, the results suggested that Dot-PAA-ELISA is a convenient, rapid, sensitive, and specific diagnostic method suitable for studying large numbers of samples obtained from clinical and epidemiological studies, thereby helping reduce important economic losses.
authors.affiliation: College of Animal Science and Veterinary Medicine, Shandong Agricultural University, No. 61, Daizong Street, Taian, 271018, Shandong, People's Republic of China.|College of Animal Science and Veterinary Medicine, Henan Institute of Science and Technology, No. 90, Hualan Street, Xinxiang, 453003, Henan, People's Republic of China.
authors.forename: Xiao-Jing
authors.initials: XJ
authors.lastname: Xia
authors.affiliation: College of Animal Science and Veterinary Medicine, Henan Institute of Science and Technology, No. 90, Hualan Street, Xinxiang, 453003, Henan, People's Republic of China.
authors.forename: Lei
authors.initials: L
authors.lastname: Wang
authors.affiliation: College of Animal Science and Veterinary Medicine, Shandong Agricultural University, No. 61, Daizong Street, Taian, 271018, Shandong, People's Republic of China. bzshenzq@163.com.|Shandong Binzhou Animal Science &Veterinary Medicine Academy, No.169, Huanghe 2th road, Binzhou, 256600, Shandong, People's Republic of China. bzshenzq@163.com.
authors.forename: Zhi-Qiang
authors.initials: ZQ
authors.lastname: Shen
authors.affiliation: Center for Experimental and Molecular Medicine, Academic Medical Center, University of Amsterdam, 1105 AZ, Amsterdam, The Netherlands.
authors.forename: Wanhai
authors.initials: W
authors.lastname: Qin
authors.affiliation: College of Animal Science and Veterinary Medicine, Henan Institute of Science and Technology, No. 90, Hualan Street, Xinxiang, 453003, Henan, People's Republic of China.
authors.forename: Jianhe
authors.initials: J
authors.lastname: Hu
authors.affiliation: College of Animal Science and Veterinary Medicine, Shandong Agricultural University, No. 61, Daizong Street, Taian, 271018, Shandong, People's Republic of China. sjjiang@sdau.edu.cn.
authors.forename: Shi-Jin
authors.initials: SJ
authors.lastname: Jiang
authors.affiliation: Shandong Binzhou Animal Science &Veterinary Medicine Academy, No.169, Huanghe 2th road, Binzhou, 256600, Shandong, People's Republic of China.
authors.forename: Shu-Guang
authors.initials: SG
authors.lastname: Li
mesh_terms: D000818:Animals; D000907:Antibodies, Bacterial / Q000097:blood*; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D005969:Glutamate Dehydrogenase / Q000276:immunology*; D011817:Rabbits; D012680:Sensitivity and Specificity; D013290:Streptococcal Infections / Q000175:diagnosis* / Q000382:microbiology / Q000662:veterinary*; D017011:Streptococcus suis / Q000276:immunology*
chemical_list: D000907:Antibodies, Bacterial; D005969:Glutamate Dehydrogenase
keywords: Diagnosis; Dot-PPA-ELISA; Streptococcus suis; rGDH