Isolation and Characterization of Avian Metapneumovirus from Broiler Breeder Chickens in Korea

Kang-Seuk Choi; Woo-Jin Jeon; Mi-Ja Park; Eun-Kyoung Lee; Jun-Hun Kwon

doi:10.4167/jbv.2009.39.4.373

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Choi, Jeon, Park, Lee, and Kwon: Isolation and Characterization of Avian Metapneumovirus from Broiler Breeder Chickens in Korea

Original Article

J Bacteriol Virol 2009;39(4):373-382.

Published online: 18 January 2009

DOI: https://doi.org/10.4167/jbv.2009.39.4.373

Isolation and Characterization of Avian Metapneumovirus from Broiler Breeder Chickens in Korea

Kang-Seuk Choi^∗,^∗∗, Woo-Jin Jeon, Mi-Ja Park, Eun-Kyoung Lee, Jun-Hun Kwon

Avian Diseases Division, National Veterinary Research and Quarantine Service, Anyang, Korea

^∗Corresponding author: Kang-Seuk Choi, DVM, MS, PhD., Avian Diseases Division, National Veterinary Research and Quarantine Service, 335 Joonangro, Manan-gu, Anyang, Gyeonggi 430-824, Korea. Phone: +82-31-467-1821, Fax: +82-31-467-1814 e-mail: choiks@nvrqs.go.kr

^∗∗ This work was supported by a grant of the National Veterinary Research and Quarantine Service (B-AD15-2008-07-01), Korea.

Revised 9 September 200919 November 2009 Accepted 24 November 2009

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

Avian metapneumovirus (AMPV) is an emerging pathogen causing respiratory and reproductive illness in poultry worldwide. To demonstrate the presence of AMPV in domestic chickens in Korea, we attempted to isolate AMPV from affected chickens. A cytopathic agent was isolated using chicken tracheal ring culture from dead chickens from a broiler breeder farm with reduced egg production in Korea. This agent, termed SC1509 strain, subsequently passed in Vero cells with distinct cytopathic effects. The SC1509 strain was confirmed as avian metapneumovirus (AMPV) using both RT-PCR test and monoclonal antibody-based immunofluorescence assay. Sequence analysis based on the G glycoprotein revealed that the SC1509 strain had 22.5 to 96.0% nucleotide sequence identity and 11.1 to 92.7% predicted amino acid sequence identity with previously published AMPV strains, particularly with the highest sequence homology (95.8 to 96% for nucleotides and 92.2 to 92.7% for amino acids) to European strains belonging to genotype B. The SC1509 strain was phylogenetically clustered with genotype B viruses, confirming that the SC1509 strain belongs to genotype B. This is the first report of genotype B avian metapneumovirus from chickens in Korea.

Keywords: Avian metapneumovirus, Genotype B, G protein, Korea

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Figure 1.

Detection of AMPV RNA in dead chickens using RT-PCR. Total RNA was isolated from nasopharyngeal swabs (A) or nasal turbinate tissues (B). Lane M, molecular marker; lanes l~6, dead chickens tested. The arrow represents the expected size of PCR products amplified by PCR.

Figure 2.

Immunofluorescent staining of AMPV (SC1509 strain)-infected Vero cells using AMPV-specific monoclonal antibody 12.2.1. A: Mock-infected Vero cells, B: AMPV-infected Vero cells.

Figure 3.

Cytopathic effects of AMPV SC1509 strain in Vero cells. A: Mock-infected Vero cells, B: AMPV-infected Vero cells.

Figure 4.

Phylogenetic anlaysis of AMPV SC1509 strain. Nucleotide sequences were analyzed based on the complete G coding region. AMPV field isolate from this study is shown in bold. Designation in parenthesis represents accession number of reference strains obtained from the GenBank database. The provisional designations including genotypes are indicated on the right.

Table 1.

Comparison of nucleic acid and deduced amino acid sequences of the G protein of AMPV SC1509 strain with other strains of AMPV

	Deduced amino acid
	1	2	3	4	5	6	7	8	9	10	11	12	13
1. SC1509	100	32.6	32.6	32.3	92.5	92.2	92.7	11.1	15.2	25.6	25.6	6.4	4.7
2. CVL14/1 (A)	51.4	100	98.4	98.2	32.6	32.6	32.6	7.8	10.3	25.3	25.3	5.8	7.5
3. 1556 (A)	51.1	99	100	99.7	32.6	32.6	32.6	8.0	10.5	25.3	25.3	5.8	7.5
4. LAH/A (A)	51.1	98.8	99.8	100	32.3	32.3	32.3	8.0	10.5	25.3	25.3	5.8	7.5
5. 2119 (B)	96.0	52.1	51.7	51.6	100	98.7	99.2	10.4	14.8	27	27	5.9	4.2
6. 872S (B)	95.8	52.2	51.8	51.7	99.4	100	99.5	10.4	14.8	27.2	27.2	5.7	4.2
7. 6574 (B)	95.9	52.3	51.9	51.9	99.5	99.6	100	10.4	14.8	27	27	5.7	4.2
8. Colorado (C)	22.5	24.5	24.7	24.6	22.1	22.2	22.2	100	52.9	12.6	12.6	18.1	9.8
9. Mn-2a (C)	35.1	38.1	38.0	37.8	35.3	35.3	35.3	61	100	15.2	15.2	10	5.8
10. Fr/85/1 (D)	45.3	52.6	52.3	52.2	45.5	45.6	45.6	25.9	37.6	100	99.7	8.1	7.7
11. Fr/85/2 (D)	45.3	52.5	52.4	52.2	45.5	45.6	45.6	25.9	37.6	99.8	100	8.1	7.7
12. HMPV	20.1	19.5	19.6	19.7	20.1	20.2	20.1	34.7	22.4	20.7	20.7	100	10.5
13. BRSV	22.3	20.3	20.4	20.3	21.7	21.8	21.6	32.8	21.2	21.2	21.3	30.6	100
						Nucleic acid

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