鸡传染性支气管炎病毒M41株全基因组序列测定及致病性分析

doi:10.11843/j.issn.0366-6964.2025.08.034

Abstract

Abstract:

In order to systematically evaluate whether the biological characteristics of infectious bronchitis virus (IBV) M41 strain will change after serial passages, we continuously passaged the IBV M41 strain (CVCC AV1511, abbreviated as AV1511-1979) introduced from France by the National Center for Veterinary Culture Collection in 1979 on SPF chickens for five times to evaluate its differences in genome variation, virus shedding, and tissue tropism. Illumina high-throughput sequencing method was used to sequence the genome sequences before and after passage, and an IBV challenge model was established. RT-PCR, RT-qPCR and tracheal ring cilia swing test were used to evaluate the differences before and after passage in tems of virus content, clinical symptom observation and tissue tropism. The results showed that the whole genome of the AV1511-1979 strain did not mutate after 5 generations of continuous passage on SPF chickens, but it was quite different from Mass strains from other sources in 1ab, 5a, 4b and N protein regions. In the virus challenge model, the inoculated chickens showed obvious clinical symptoms such as depression, mouth breathing and respiratory rales. The ciliary activity of the tracheal ring was examined by autopsy, and it was found that the ciliary activity was poor or stopped. The results of RT-qPCR showed that the virus could be replicated in the trachea, lung, kidney, glandular stomach, duodenum and cecal tonsil of infected chickens, especially in the trachea, kidney and glandular stomach. There was no significant difference in viral load before and after passage. The results showed that the biological characteristics of the IBV M41 strain, AV1511-1979, were stable and did not change within 5 generations on SPF chickens. As a standard virulent strain for the production of IBV inactivated vaccine and IBV vaccine testing, our work provides a reference for the generation control of production viruses and vaccine evaluation.

Key words: avian infectious bronchitis virus, M41 strain, SPF chickens, passage, biological characteristics

CLC Number:

S852.659.6

ZHAO Jingyu, LI Dan, ZHANG Bing, ZHANG Qianyi, ZHANG Jinhua, SONG Yafen, YANG Chenghuai. Genome Sequencing and Pathogenicity Analysis of Infectious Bronchitis Virus M41 Strain[J]. Acta Veterinaria et Zootechnica Sinica, 2025, 56(8): 3967-3975.

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References 29

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引物名称 Primer names	引物序列(5′→3′) Primer sequences (5′→3′)	片段大小/bp Size
IBV-F	ATGTCTATCGCCAGGGAAATGTC	266
IBV-R	GCTCTAACTCTATACTAGCCTA
Mass-F	AGTTTCAATTGCTTACGGTCCT	360
Mass-R	AATAGCCAAACCTGCGTCT

编号 No.	GenBank登录号 GenBank accession No.	编号No.
编号 No.	GenBank登录号 GenBank accession No.	1	2	3	4	5
1	AV1511-1979	*	99.9	92.2	92.2	92.2
2	ON713866.1	99.6	*	92.2	92.2	92.1
3	DQ834384	97.8	97.6	*	99.9	99.9
4	MK937830	97.6	97.5	99.0	*	99.9
5	AY851295	97.8	97.6	100.0	99.9	*

编码区 CDS	最大同一性 The most percent identity
1a	89.1
5a	89.4
N蛋白	90.7
1b	93.5
3a	94.8
5b	95.2
m蛋白	96.2
E蛋白	96.9
S蛋白	97.7
3b	99.5

毒株 Strain	临床观察结果^a Clinical observation results						血清转阳结果^d Results of seroconversion
毒株 Strain	3 dpi	5 dpi	7 dpi	9 dpi	11 dpi	13 dpi	14 dpi
F1-E1 AV1511-1979	18/30^b (60%)^c	26/27 (96%)	24/24 (100%)	14/14 (100%)	7/14 (50%)	3/14 (21%)	3/14 (21%)
F5-E1 AV1511-1979	20/30 (67%)	25/27 (93%)	20/24 (83%)	14/14 (100%)	7/14 (50%)	3/14 (21%)	5/14 (36%)

毒株 Strain	F1-E1 AV1511-1979			F5-E1 AV1511-1979
毒株 Strain	前段 Proximal segment	中段 Middle segment	后段 Distal segment	前段 Proximal segment	中段 Middle segment	后段 Distal segment
1	0、0、0	0、0、0、0	0、0、0	0、0、0	0、0、0、0	0、0、0
2	2、2、0	0、0、0、0	1、0、0	0、0、0	0、0、0、0	0、0、0
3	0、0、0	0、0、0、0	0、0、0	0、0、0	0、0、0、0	0、0、0
4	1、0、0	0、0、0、0	0、0、0	0、0、0	0、0、0、0	0、0、0
5	0、0、0	0、0、0、0	0、0、0	0、0、0	0、0、0、0	0、0、0
6	1、1、1	0、0、1、1	2、1、0	0、0、0	0、0、0、0	0、0、0
7	0、0、1	1、1、2、0	2、2、0	0、0、0	0、0、0、0	0、0、0
8	0、0、0	1、0、0、0	1、1、0	0、0、0	0、0、0、0	0、0、0

Genome Sequencing and Pathogenicity Analysis of Infectious Bronchitis Virus M41 Strain

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