PMID- 30442149 OWN - NLM STAT- MEDLINE DCOM- 20181227 LR - 20181227 IS - 1746-6148 (Electronic) IS - 1746-6148 (Linking) VI - 14 IP - 1 DP - 2018 Nov 15 TI - A multiplex xTAG assay for the simultaneous detection of five chicken immunosuppressive viruses. PG - 347 LID - 10.1186/s12917-018-1663-1 [doi] LID - 347 AB - BACKGROUND: Chicken anemia virus (CAV), avian reovirus (ARV), infectious bursal disease virus (IBDV), Marek's disease virus (MDV) and reticuloendotheliosis virus (REV) all cause immunosuppressive disease in birds through vertical or horizontal transmission. Mixed infections with these immunosuppressive pathogens lead to atypical clinical signs and obstruct accurate diagnoses and epidemiological investigations. Therefore, it is essential to develop a high-throughput assay for the simultaneous detection of these immunosuppressive viruses with high specificity and sensitivity. The aim of this study was to establish a novel method using a RT-PCR assay combined with fluorescence labeled polystyrene bead microarray (multiplex xTAG assay) to detect single or mixed viral infections. RESULTS: The results showed that the established xTAG assay had no nonspecific reactions with avian influenza virus (AIV), infectious bronchitis virus (IBV), newcastle disease virus (NDV), infectious laryngotracheitis virus (ILTV), Mycoplasma gallisepticum (MG) and Mycoplasma synoviae (MS). The limit of detection was 1.0 x 10(3) copies/muL for IBDV and 1.0 x 10(2)copies/muL for the other four viruses. Ninety field samples were tested and the results were confirmed using conventional RT-PCR methods. The detection results of these two methods were 100% consistent. The established multiplex xTAG assay allows a high throughput and simultaneous detection of five chicken immunosuppressive viruses. CONCLUSION: The multiplex xTAG assay has been showed to be an additional tool for molecular epidemiology studies of five chicken immunosuppressive viruses in the poultry industry. FAU - Cong, Feng AU - Cong F AD - Guangdong Laboratory Animals Monitoring Institute and Guangdong Provincial Key Laboratory of Laboratory Animals, Guangzhou, 510633, China. FAU - Zhu, Yujun AU - Zhu Y AD - Guangdong Laboratory Animals Monitoring Institute and Guangdong Provincial Key Laboratory of Laboratory Animals, Guangzhou, 510633, China. FAU - Wang, Jing AU - Wang J AD - Guangdong Laboratory Animals Monitoring Institute and Guangdong Provincial Key Laboratory of Laboratory Animals, Guangzhou, 510633, China. FAU - Lian, Yuexiao AU - Lian Y AD - Guangdong Laboratory Animals Monitoring Institute and Guangdong Provincial Key Laboratory of Laboratory Animals, Guangzhou, 510633, China. AD - Guangdong Provincial Key Laboratory of Zoonosis Prevention and Control, College of Veterinary Medicine, South China Agricultural University, Guangzhou, 510640, China. FAU - Liu, Xiangnan AU - Liu X AD - Guangdong Laboratory Animals Monitoring Institute and Guangdong Provincial Key Laboratory of Laboratory Animals, Guangzhou, 510633, China. AD - Guangdong Provincial Key Laboratory of Zoonosis Prevention and Control, College of Veterinary Medicine, South China Agricultural University, Guangzhou, 510640, China. FAU - Xiao, Li AU - Xiao L AD - Guangdong Laboratory Animals Monitoring Institute and Guangdong Provincial Key Laboratory of Laboratory Animals, Guangzhou, 510633, China. FAU - Huang, Ren AU - Huang R AD - Guangdong Laboratory Animals Monitoring Institute and Guangdong Provincial Key Laboratory of Laboratory Animals, Guangzhou, 510633, China. FAU - Zhang, Yu AU - Zhang Y AD - Guangdong Laboratory Animals Monitoring Institute and Guangdong Provincial Key Laboratory of Laboratory Animals, Guangzhou, 510633, China. FAU - Chen, Meili AU - Chen M AD - Guangdong Laboratory Animals Monitoring Institute and Guangdong Provincial Key Laboratory of Laboratory Animals, Guangzhou, 510633, China. chml@gdlami.com. FAU - Guo, Pengju AU - Guo P AD - Guangdong Laboratory Animals Monitoring Institute and Guangdong Provincial Key Laboratory of Laboratory Animals, Guangzhou, 510633, China. vetbio2016@hotmail.com. LA - eng GR - 2015A030313693/the Science and Technology Department of Guangdong Province and Guangzhou/ GR - 2017B030314171/the Science and Technology Department of Guangdong Province and Guangzhou/ GR - 2016201604030059/the Science and Technology Department of Guangdong Province and Guangzhou/ GR - 2017A070702001/the Science and Technology Department of Guangdong Province and Guangzhou/ GR - 2015BAI07B01/National Key Technology R&D Program/ GR - 2016A030303024/Department of Science and Technology of Guangdong Province and Guangzhou Municipa/ PT - Journal Article DEP - 20181115 PL - England TA - BMC Vet Res JT - BMC veterinary research JID - 101249759 SB - IM MH - Animals MH - Birnaviridae Infections/diagnosis/*veterinary/virology MH - *Chicken anemia virus MH - Chickens/virology MH - Circoviridae Infections/diagnosis/*veterinary/virology MH - Coinfection/diagnosis/*veterinary/virology MH - *Infectious bursal disease virus MH - *Mardivirus MH - Marek Disease/*diagnosis/virology MH - Microarray Analysis/methods/*veterinary MH - Multiplex Polymerase Chain Reaction/methods/*veterinary MH - *Orthoreovirus, Avian MH - Poultry Diseases/*diagnosis/virology MH - Reoviridae Infections/diagnosis/*veterinary/virology MH - Reproducibility of Results MH - *Reticuloendotheliosis Viruses, Avian MH - Retroviridae Infections/diagnosis/*veterinary/virology MH - Sensitivity and Specificity MH - Tumor Virus Infections/diagnosis/*veterinary/virology PMC - PMC6238339 OTO - NOTNLM OT - Avian reovirus OT - Chicken anemia virus OT - Chicken immunosuppressive viruses OT - Multiplex xTAG assay OT - Simultaneous detection COIS- ETHICS APPROVAL AND CONSENT TO PARTICIPATE: Not applicable. CONSENT FOR PUBLICATION: Not applicable. COMPETING INTERESTS: The authors declare that they have no competing interests. PUBLISHER'S NOTE: Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations. EDAT- 2018/11/18 06:00 MHDA- 2018/12/28 06:00 PMCR- 2018/11/15 CRDT- 2018/11/17 06:00 PHST- 2018/06/14 00:00 [received] PHST- 2018/10/22 00:00 [accepted] PHST- 2018/11/17 06:00 [entrez] PHST- 2018/11/18 06:00 [pubmed] PHST- 2018/12/28 06:00 [medline] PHST- 2018/11/15 00:00 [pmc-release] AID - 10.1186/s12917-018-1663-1 [pii] AID - 1663 [pii] AID - 10.1186/s12917-018-1663-1 [doi] PST - epublish SO - BMC Vet Res. 2018 Nov 15;14(1):347. doi: 10.1186/s12917-018-1663-1.