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Avian Diseases 50(3):386-390. 2006
doi: http://dx.doi.org/10.1637/7502-010906R1.1

Isolation, Identification, and Comparison of Four Isolates of Avian Paramyxovirus Serotype 2 in China

Guo-zhong Zhang, Ji-xun ZhaoA , Hong-wei Wang, Ai-mei Yang, Chun-ya Bu, and Ming WangA

Department of Preventive Veterinary Medicine, College of Veterinary Medicine, China Agricultural University, Beijing 100094, P.R. China

^ACorresponding authors

Abstract

Four Yucaipa-like viruses of avian paramyxovirus serotype 2 (APMV-2) were isolated in China from the imported Gouldian Finch (Chloebia gouldiae) and broilers in 1998–2002, and were named F4, F6, F8, and NK, respectively. Examined under electron microscope, the isolates were found to be round in shape and varying in size. The results of the hemagglutination inhibition test and indirect enzyme-linked immunosorbent assay (using monoclonal antibodies) showed some differences between the isolates and the reference strain Yucaipa. The isolates derived from chickens had a closer relationship to Yucaipa virus than did those of finches. Sequence comparison of the fusion gene and the haemagglutinin-neuraminidase gene showed similar results, although the variations were lesser among APMV-2 viruses in nucleotide and amino acid sequence. By sequence comparison, it was also revealed that at the molecular level the four virus strains belong to APMV-2, and that two of the strains were isolated from the same group of imported Gouldian Finches.

Resumen

Aislamiento, identificación y comparación de cuatro aislamientos de paramixovirus aviar serotipo 2 en China.

En China, durante el periodo comprendido entre 1998 y 2002 se aislaron cuatro virus parecidos al virus Yucaipa perteneciente al serotipo 2 del paramixovirus aviar, procedentes de pinzones de gould (Chloebia gouldiae) importados y de pollos de engorde. Los aislamientos fueron identificados como F4, F6, F8, y NK, respectivamente. Observados al microscopio electrónico, los aislamientos resultaron ser de forma redondeada y de tamaño variable. Los resultados de la prueba de inhibición de la hemoaglutinación y de la prueba de inmunoensayo asociado a enzimas (utilizando anticuerpos monoclonales) demostraron algunas diferencias entre los aislamientos y la cepa de referencia Yucaipa. Los aislamientos derivados de pollos de engorde mostraron una relación más cercana con el virus Yucaipa que la mostrada por los virus provenientes de los pinzones. La comparación de la secuencia del gen de fusión y del gen de la hemaglutinina–neuraminidasa mostró similitudes entre las cepas, aunque las variaciones en la secuencia de nucleótidos y de aminoácidos fueron menores entre los paramixovirus serotipo 2. Mediante la comparación de las secuencias se reveló que a nivel molecular las cuatro cepas pertenecen al serotipo 2 del paramixovirus aviar y que dos de las cepas se aislaron del mismo grupo de pinzones de gould importados.

Abbreviations: APMV-2 = avian paramyxovirus serotype 2; CRBCs = chicken red blood cells; ELISA = enzyme-linked immunosorbent assay; EM = electron microscopy; F = fusion; HA = hemagglutination; HI = hemagglutination inhibition; HN = hemagglutinin-neuraminidase; IEM = immune electron microscopy; McAb = monoclonal antibody; PBS = phosphate-buffered saline; RT-PCR = reverse transcriptase–polymerase chain reaction; SPF = specific-pathogen-free; TEM = transmission electron microscopy

Received: 9 January 2006; Accepted: 20 March 2006

Keywords: avian paramyxovirus serotype 2, Chinese isolates, comparison, sequence analysis

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Fig. 1. Electron micrograph of APMV-2 isolates (33,000×). The picture shows APMV-2 virus-like particles of approximately 150–300 nm in diameter (arrows). A, C, E, and G are TEM micrographs; B, D, F, and H are IEM micrographs; A and B are micrographs of F4 strain; C and D are micrographs of F6 strain; E and F are micrographs of F8 strain; G and H are micrographs of standard-strain Yucaipa

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Fig. 2. Phylogenetic tree of HN gene. GenBank accession numbers AF515837, AF515839, AF515838, and D14030

Table 1. Primers used for PCR amplification and sequence analysis

Table 2. Crossing HI relationships between the isolates and the standard virus (log 2)

Table 3. Antigenicity difference analysis of the isolates using McAbs to APMV-2

Table 4. Nucleotide identity of APMV-2 isolates to reference strains Yucaipa in the HN and F segments

Table 5. Amino acid identity of APMV-2 isolates to reference strains Yucaipa in the HN and F segments

Table 6. Amino acid changes of HN and F genes at all polymorphic locations

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