Detection of Infectious Bursal Disease Virus by Double Antibody Sandwich ELISA

Woo-Jin Jeon; Byung-Sik Chang; Mi-Ja Park; Eun-Kyoung Lee; Hoo-Don Joo; Jun-Hun Kwon; Kang-Seuk Choi

doi:10.4167/jbv.2008.38.3.139

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Jeon, Chang, Park, Lee, Joo, Kwon, and Choi: Detection of Infectious Bursal Disease Virus by Double Antibody Sandwich ELISA

Original Article

J Bacteriol Virol 2008;38(3):139-147.

Published online: 18 January 2008

DOI: https://doi.org/10.4167/jbv.2008.38.3.139

Detection of Infectious Bursal Disease Virus by Double Antibody Sandwich ELISA

Woo-Jin Jeon¹, Byung-Sik Chang², Mi-Ja Park¹, Eun-Kyoung Lee¹, Hoo-Don Joo², Jun-Hun Kwon¹, Kang-Seuk Choi^1,^∗

¹National Veterinary Research Institute, National Veterinary Research and Quarantine ServiceAnyang, Gyeonggi, Republic of Korea

²JenoBiotech Inc, Chuncheon, Gangwon, Republic of Korea

^∗Corresponding author: Kang-Seuk Choi, DVM, MS, PhD. National Veterinary Research Institute, National Veterinary Research and Quarantine Service, Anyang, Gyeonggi, 430-824, Republic of Korea. Phone: +82-31-467-1821, Fax: +82-31-467-1814 e-mail: choiks@nvrqs.go.kr

^∗∗ This work was supported by a grant (M-AD15-2007-07-01) from National Veterinary Research & Quarantine Service.

Revised 16 June 200825 August 2008 Accepted 1 September 2008

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

Abstract

Infectious bursal disease virus (IBDV) is responsible for a highly contagious disease of poultry causing severe immunosuppression in chickens. A double antibody sandwich ELISA (DAS-ELISA) was developed to detect IBDV from clinical samples. Two kinds of anti-IBDV antibodies, monoclonal antibody R63 and chicken anti-IBDV sera, were used for DAS-ELISA. Detection limit of IBDV by DAS-ELISA was approximately 10^2.7 EID₅₀/ml. The DAS-ELISA detected IBDV from most (13/14) of vaccine products including mild, intermediate and intermediate-plus types. The DAS-ELISA also detected IBDV from all (19/19) of field Korean isolates including very virulent and intermediate-plus phenotypes. Our results indicate that the DAS-ELISA would provide useful diagnostic tool to detect IBDV from clinical samples as well as rapid quantitative detection of IBDV.

Keywords: Antigen detection, Diagnosis, ELISA, Infectious bursal disease

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Figure 1.

Quantitative titration of IBDV D67 strain by DAS ELISA (B) and comparison with RT-PCR assay (A).

Figure 2.

Detection of IBDV in commercial IBDV vaccines by DAS-ELISA (B) and comparison with results of RT-PCR (A). Data in the RT-PCR (A) represents relative quantity percent of each amplified DNA to total DNA amplified in the gel.

Figure 3.

Detection of field isolates of IBDV by DAS-ELISA. Line (OD 0.53 at 450 nm) represents cut-off.

Table 1.

Field IBDV strains used for evaluation of DAS-ELISA

Virus Sample	IBDV Strain	Phenotype^a	History
Virus Sample	IBDV Strain	Phenotype^a	Region	Year	Breed^b	Passage^c
VV1	SH92	VV	Chungnam	1992	L	CE5
VV2	07D29	VV	Chungnam	2007	B	CE1
VV3	06Q44	VV	Chungbuk	2006	B	CE2
VV4	06D62	VV	Gangwon	2006	B	CE1
VV5	05Q66	VV	Chungnam	2005	B	CE2
VV6	07D4	VV	Chungbuk	2007	B	CE1
VV7	05D99	VV	Gyeonggi	2005	KNB	CE2
VV8	07D6	VV	Jeonbuk	2007	B	CE1
VV9	05D62	VV	Jeonnam	2005	B	CE2
VV10	04Q10	VV	Jeonnam	2004	B	BF
VV11	03099	VV	Gyeonggi	2003	B	CE2
VV12	07D11	VV	Gyeonggi	2007	B	CE1
IP1	03243	I+	Jeonnam	2003	B	CE2
IP2	07D24	I+	Chungbuk	2007	B	BF
IP3	05D110	I+	Chungnam	2005	L	CE2
IP4	05D58	I+	Chungbuk	2005	B	CE1
IP5	05D42	I+	Jeonbuk	2005	B	BF
IP6	03244	I+	Jeonbuk	2003	B	CE2
IP7	05D21	I+	Gyeonggi	2005	B	CT

^a V V, very virulent; I+, intermediate plus. The phenotype was determined by phylogenetic analysis based on the VP2 protein gene of IBDV.

^b B, broiler; L, layer; KNB, Korean native boiler

^c CE, chicken embryo (passage no.); BF, bursa of Fabricius; CT, cecal tonsil

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