A New Strategy for the Detection of Chicken Infectious Anemia Virus Contamination in Attenuated Live Vaccine by Droplet Digital PCR

Chicken infectious anemia virus (CIAV) causes the atrophy of bone marrow hematopoietic and lymphoid tissues in chicks, leading to huge economic losses all over the world. The using of attenuated vaccine contaminated with CIAV increased the mortality and the pathogenicity of other diseases in many fa...

Full description

Bibliographic Details
Main Authors: Qiuchen Li, Yubiao Zhang, Fanfeng Meng, Hui Jiang, Guanlong Xu, Jiabo Ding, Yawen Zhang, Guiwei Dong, Sibao Tian, Shuang Chang, Peng Zhao
Format: Article
Language:English
Published: Hindawi Limited 2019-01-01
Series:BioMed Research International
Online Access:http://dx.doi.org/10.1155/2019/2750472
id doaj-d13593b6c28040e9be0d451027137d6b
record_format Article
spelling doaj-d13593b6c28040e9be0d451027137d6b2020-11-25T01:37:50ZengHindawi LimitedBioMed Research International2314-61332314-61412019-01-01201910.1155/2019/27504722750472A New Strategy for the Detection of Chicken Infectious Anemia Virus Contamination in Attenuated Live Vaccine by Droplet Digital PCRQiuchen Li0Yubiao Zhang1Fanfeng Meng2Hui Jiang3Guanlong Xu4Jiabo Ding5Yawen Zhang6Guiwei Dong7Sibao Tian8Shuang Chang9Peng Zhao10College of Veterinary Medicine, Shandong Agricultural University, Tai’an, Shandong, ChinaYuyao Animal Husbandry and Veterinary Bureau, Ningbo, Zhejiang, ChinaBeijing Dafaun Poultry Breeding Company Ltd., Beijing, ChinaNational Reference Laboratory for Animal Brucellosis, China Institute of Veterinary Drug Control, Beijing 102629, ChinaNational Reference Laboratory for Animal Brucellosis, China Institute of Veterinary Drug Control, Beijing 102629, ChinaNational Reference Laboratory for Animal Brucellosis, China Institute of Veterinary Drug Control, Beijing 102629, ChinaCollege of Veterinary Medicine, Shandong Agricultural University, Tai’an, Shandong, ChinaCollege of Veterinary Medicine, Shandong Agricultural University, Tai’an, Shandong, ChinaCollege of Veterinary Medicine, Shandong Agricultural University, Tai’an, Shandong, ChinaCollege of Veterinary Medicine, Shandong Agricultural University, Tai’an, Shandong, ChinaCollege of Veterinary Medicine, Shandong Agricultural University, Tai’an, Shandong, ChinaChicken infectious anemia virus (CIAV) causes the atrophy of bone marrow hematopoietic and lymphoid tissues in chicks, leading to huge economic losses all over the world. The using of attenuated vaccine contaminated with CIAV increased the mortality and the pathogenicity of other diseases in many farms. However, it is difficult to detect the CIAV contamination by general detection technology due to the extremely low dose of CIAV in vaccines. In this study, we established a new method called droplet digital Polymerase Chain Reaction (ddPCR) to detect CIAV contamination of vaccines more sensitively and accurately. The lowest detection limitation of this method is 2.4 copies of CIAV plasmid or CIAV contamination at 0.1 EID50/1000 feathers in vaccines without any positive signals of other viruses. Besides, the sensitivity of ddPCR is 100 times greater than that of conventional PCR and 10 times greater than that of real-time PCR. The ddPCR technique is more sensitive and more intuitive. Therefore, it could be valuable for the detection of CIAV contamination in vaccines.http://dx.doi.org/10.1155/2019/2750472
collection DOAJ
language English
format Article
sources DOAJ
author Qiuchen Li
Yubiao Zhang
Fanfeng Meng
Hui Jiang
Guanlong Xu
Jiabo Ding
Yawen Zhang
Guiwei Dong
Sibao Tian
Shuang Chang
Peng Zhao
spellingShingle Qiuchen Li
Yubiao Zhang
Fanfeng Meng
Hui Jiang
Guanlong Xu
Jiabo Ding
Yawen Zhang
Guiwei Dong
Sibao Tian
Shuang Chang
Peng Zhao
A New Strategy for the Detection of Chicken Infectious Anemia Virus Contamination in Attenuated Live Vaccine by Droplet Digital PCR
BioMed Research International
author_facet Qiuchen Li
Yubiao Zhang
Fanfeng Meng
Hui Jiang
Guanlong Xu
Jiabo Ding
Yawen Zhang
Guiwei Dong
Sibao Tian
Shuang Chang
Peng Zhao
author_sort Qiuchen Li
title A New Strategy for the Detection of Chicken Infectious Anemia Virus Contamination in Attenuated Live Vaccine by Droplet Digital PCR
title_short A New Strategy for the Detection of Chicken Infectious Anemia Virus Contamination in Attenuated Live Vaccine by Droplet Digital PCR
title_full A New Strategy for the Detection of Chicken Infectious Anemia Virus Contamination in Attenuated Live Vaccine by Droplet Digital PCR
title_fullStr A New Strategy for the Detection of Chicken Infectious Anemia Virus Contamination in Attenuated Live Vaccine by Droplet Digital PCR
title_full_unstemmed A New Strategy for the Detection of Chicken Infectious Anemia Virus Contamination in Attenuated Live Vaccine by Droplet Digital PCR
title_sort new strategy for the detection of chicken infectious anemia virus contamination in attenuated live vaccine by droplet digital pcr
publisher Hindawi Limited
series BioMed Research International
issn 2314-6133
2314-6141
publishDate 2019-01-01
description Chicken infectious anemia virus (CIAV) causes the atrophy of bone marrow hematopoietic and lymphoid tissues in chicks, leading to huge economic losses all over the world. The using of attenuated vaccine contaminated with CIAV increased the mortality and the pathogenicity of other diseases in many farms. However, it is difficult to detect the CIAV contamination by general detection technology due to the extremely low dose of CIAV in vaccines. In this study, we established a new method called droplet digital Polymerase Chain Reaction (ddPCR) to detect CIAV contamination of vaccines more sensitively and accurately. The lowest detection limitation of this method is 2.4 copies of CIAV plasmid or CIAV contamination at 0.1 EID50/1000 feathers in vaccines without any positive signals of other viruses. Besides, the sensitivity of ddPCR is 100 times greater than that of conventional PCR and 10 times greater than that of real-time PCR. The ddPCR technique is more sensitive and more intuitive. Therefore, it could be valuable for the detection of CIAV contamination in vaccines.
url http://dx.doi.org/10.1155/2019/2750472
work_keys_str_mv AT qiuchenli anewstrategyforthedetectionofchickeninfectiousanemiaviruscontaminationinattenuatedlivevaccinebydropletdigitalpcr
AT yubiaozhang anewstrategyforthedetectionofchickeninfectiousanemiaviruscontaminationinattenuatedlivevaccinebydropletdigitalpcr
AT fanfengmeng anewstrategyforthedetectionofchickeninfectiousanemiaviruscontaminationinattenuatedlivevaccinebydropletdigitalpcr
AT huijiang anewstrategyforthedetectionofchickeninfectiousanemiaviruscontaminationinattenuatedlivevaccinebydropletdigitalpcr
AT guanlongxu anewstrategyforthedetectionofchickeninfectiousanemiaviruscontaminationinattenuatedlivevaccinebydropletdigitalpcr
AT jiaboding anewstrategyforthedetectionofchickeninfectiousanemiaviruscontaminationinattenuatedlivevaccinebydropletdigitalpcr
AT yawenzhang anewstrategyforthedetectionofchickeninfectiousanemiaviruscontaminationinattenuatedlivevaccinebydropletdigitalpcr
AT guiweidong anewstrategyforthedetectionofchickeninfectiousanemiaviruscontaminationinattenuatedlivevaccinebydropletdigitalpcr
AT sibaotian anewstrategyforthedetectionofchickeninfectiousanemiaviruscontaminationinattenuatedlivevaccinebydropletdigitalpcr
AT shuangchang anewstrategyforthedetectionofchickeninfectiousanemiaviruscontaminationinattenuatedlivevaccinebydropletdigitalpcr
AT pengzhao anewstrategyforthedetectionofchickeninfectiousanemiaviruscontaminationinattenuatedlivevaccinebydropletdigitalpcr
AT qiuchenli newstrategyforthedetectionofchickeninfectiousanemiaviruscontaminationinattenuatedlivevaccinebydropletdigitalpcr
AT yubiaozhang newstrategyforthedetectionofchickeninfectiousanemiaviruscontaminationinattenuatedlivevaccinebydropletdigitalpcr
AT fanfengmeng newstrategyforthedetectionofchickeninfectiousanemiaviruscontaminationinattenuatedlivevaccinebydropletdigitalpcr
AT huijiang newstrategyforthedetectionofchickeninfectiousanemiaviruscontaminationinattenuatedlivevaccinebydropletdigitalpcr
AT guanlongxu newstrategyforthedetectionofchickeninfectiousanemiaviruscontaminationinattenuatedlivevaccinebydropletdigitalpcr
AT jiaboding newstrategyforthedetectionofchickeninfectiousanemiaviruscontaminationinattenuatedlivevaccinebydropletdigitalpcr
AT yawenzhang newstrategyforthedetectionofchickeninfectiousanemiaviruscontaminationinattenuatedlivevaccinebydropletdigitalpcr
AT guiweidong newstrategyforthedetectionofchickeninfectiousanemiaviruscontaminationinattenuatedlivevaccinebydropletdigitalpcr
AT sibaotian newstrategyforthedetectionofchickeninfectiousanemiaviruscontaminationinattenuatedlivevaccinebydropletdigitalpcr
AT shuangchang newstrategyforthedetectionofchickeninfectiousanemiaviruscontaminationinattenuatedlivevaccinebydropletdigitalpcr
AT pengzhao newstrategyforthedetectionofchickeninfectiousanemiaviruscontaminationinattenuatedlivevaccinebydropletdigitalpcr
_version_ 1725057078024732672