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Bead-based suspension array for simultaneous differential detection of five major swine viruses

文献类型： 外文期刊

作者： Chen, Ru 1 ; Yu, Xiao-Lu 2 ; Gao, Xiao-Bo 3 ; Xue, Cun-Yi 2 ; Song, Chang-Xu 4 ; Li, Yan 4 ; Cao, Yong-Chang 2 ;

作者机构： 1.Guangdong Entry Exit Inspect & Quarantine Bur, Ctr Tech, Anim Inspect & Quarantine Lab, Guangzhou 510623, Guangdong, Peoples R China

2.Sun Yat Sen Univ, Sch Life Sci, Guangzhou 510275, Guangdong, Peoples R China

3.Natl Res Inst Family Planning, Dept Genet, Beijing 100081, Peoples R China

4.Guangdong Acad Agr Sci, Vet Med Inst, Guangzhou, Guangdong, Peoples R China

关键词： xMAP technology;Bead-based suspension array;Swine virus;Multiplex detection

期刊名称：APPLIED MICROBIOLOGY AND BIOTECHNOLOGY （ 影响因子：4.813； 五年影响因子：4.697 ）

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收录情况： SCI

摘要： 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 2.2-22 copies/mu L (corresponding to 0.5-6.8 fg/mu L 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 2 h. 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.

  • 相关文献

[1]Novel multiplex PCR assay using locked nucleic acid (LNA)-based universal primers for the simultaneous detection of five swine viruses. Chen, Ru,Qiu, Yang,Gao, Xiao-Bo,Yu, Xiao-Lu,Song, Chang-Xu.

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