caa a22 4500 605505241 CHVBK 20210128100621.0 cr unu---uuuuu 210128e20150101xx s 000 0 eng 10.1007/s00253-014-6337-8 doi (NATIONALLICENCE)springer-10.1007/s00253-014-6337-8 Bead-based suspension array for simultaneous differential detection of five major swine viruses [Elektronische Daten] [Ru Chen, Xiao-Lu Yu, Xiao-Bo Gao, Cun-Yi Xue, Chang-Xu Song, Yan Li, Yong-Chang Cao] A novel multiplex detection array based on Luminex xMAP technology was developed and validated for simultaneous detection of five major viruses causing swine reproductive diseases. By combining one-step asymmetric multiplex reverse transcription polymerase chain reaction (RT-PCR) with xMAP bead-based hybridization and flow cytometry analysis, the resulting multiplex assay was capable of detecting single and mixed infections of PRRSV, PCV-2, PRV, CSFV, and PPV in a single reaction. The assay accurately detected and differentiated 23 viral strains used in this study. The low detection limit was determined as 2.2-22copies/μL (corresponding to 0.5-6.8fg/μL DNA template) on plasmid constructs containing viral fragments. The intra-assay and inter-assay variances (CV%) were low that ranged from 2.5 to 5.4% and 4.1 to 7.6%, respectively. The assay was applied to test field samples and detected single and mixed viral infections. The detection rate was higher than that of uniplex conventional PCR and RT-PCR methods. The detection of PRRSV by the bead-based multiplex assay was comparable with a commercially available real time RT-PCR kit. The test procedure on purified DNA or RNA samples could be completed within 2h. In conclusion, the bead-based suspension array presented here proved to be a high-throughput practical tool that provided highly specific and sensitive identification of single and multiple infections of five major viruses in pigs and boar semen. Springer-Verlag Berlin Heidelberg, 2015 xMAP technology nationallicence Bead-based suspension array nationallicence Swine virus nationallicence Multiplex detection nationallicence Chen Ru Animal Inspection and Quarantine Laboratory, Technical Center, Guangdong Entry-Exit Inspection and Quarantine Bureau, No.66 Huacheng Dadao Ave., Zhujiangxincheng, 510623, Guangzhou, China aut Yu Xiao-Lu School of Life Sciences, Sun Yat-Sen University, Guangzhou, China aut Gao Xiao-Bo Department of Genetics, National Research Institute for Family Planning, 100081, Beijing, China aut Xue Cun-Yi School of Life Sciences, Sun Yat-Sen University, Guangzhou, China aut Song Chang-Xu Veterinary Medicine Institute, Guangdong Academy of Agricultural Sciences, Guangzhou, China aut Li Yan Veterinary Medicine Institute, Guangdong Academy of Agricultural Sciences, Guangzhou, China aut Cao Yong-Chang School of Life Sciences, Sun Yat-Sen University, Guangzhou, China aut Applied Microbiology and Biotechnology Springer Berlin Heidelberg 99/2(2015-01-01), 919-928 0175-7598 99:2<919 2015 99 253 https://doi.org/10.1007/s00253-014-6337-8 text/html Onlinezugriff via DOI BK010053 XK010053 XK010000 Metadata rights reserved Springer special CC-BY-NC licence nationallicence 1 research-article jats NATIONALLICENCE NATIONALLICENCE NL-springer NATIONALLICENCE 856 40 https://doi.org/10.1007/s00253-014-6337-8 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Chen Ru Animal Inspection and Quarantine Laboratory, Technical Center, Guangdong Entry-Exit Inspection and Quarantine Bureau, No.66 Huacheng Dadao Ave., Zhujiangxincheng, 510623, Guangzhou, China aut NATIONALLICENCE 700 1- Yu Xiao-Lu School of Life Sciences, Sun Yat-Sen University, Guangzhou, China aut NATIONALLICENCE 700 1- Gao Xiao-Bo Department of Genetics, National Research Institute for Family Planning, 100081, Beijing, China aut NATIONALLICENCE 700 1- Xue Cun-Yi School of Life Sciences, Sun Yat-Sen University, Guangzhou, China aut NATIONALLICENCE 700 1- Song Chang-Xu Veterinary Medicine Institute, Guangdong Academy of Agricultural Sciences, Guangzhou, China aut NATIONALLICENCE 700 1- Li Yan Veterinary Medicine Institute, Guangdong Academy of Agricultural Sciences, Guangzhou, China aut NATIONALLICENCE 700 1- Cao Yong-Chang School of Life Sciences, Sun Yat-Sen University, Guangzhou, China aut NATIONALLICENCE 773 0- Applied Microbiology and Biotechnology Springer Berlin Heidelberg 99/2(2015-01-01), 919-928 0175-7598 99:2<919 2015 99 253