胸膜肺炎放线杆菌分离株血清型的PCR鉴定和Apx毒素基因检测

doi:10.11843/j.issn.0366-6964.2023.03.044

畜牧兽医学报 ›› 2023, Vol. 54 ›› Issue (3): 1341-1346.doi: 10.11843/j.issn.0366-6964.2023.03.044

• 研究简报 • 上一篇

胸膜肺炎放线杆菌分离株血清型的PCR鉴定和Apx毒素基因检测

卢碧凯^1,, 袁秀芳², 徐丽华², 余斌², 苏菲², 叶十一², 陈怡洁^1,2, 蒋利明³, 张晖^1*, 李军星^2*

1. 佛山科学技术学院生命科学与工程学院, 佛山 528200;
2. 浙江省农业科学院畜牧兽医研究所, 杭州 310021;
3. 浙江省台州市动物疫病预防控制中心, 台州 317700

收稿日期:2022-06-27 出版日期:2023-03-23 发布日期:2023-03-21
通讯作者: 张晖,主要从事动物组织学与胚胎学研究,E-mail:zhanghui429@hotmail.com;李军星,主要从事猪细菌病的防治研究,E-mail:lijunx@zaas.ac.cn
作者简介:卢碧凯(1995-)，男，壮族，广西南宁人，硕士生，主要从事动物疫病防控和兽医公共卫生研究，E-mail: 710817451@qq.com
基金资助:
浙江省自然科学基金(LY21C180002)；浙江省重点研发计划(2019C02052-3)；台州市农业科技项目

Molecular Serotyping and Apx Gene Profile Analysis of Actinobacillus pleuropneumoniae Isolates

LU Bikai^1,, YUAN Xiufang², XU Lihua², YU Bin², SU Fei², YE Shiyi², CHEN Yijie^1,2, JIANG Liming³, ZHANG Hui^1*, LI Junxing^2*

1. School of Life Science and Technology, Foshan University, Foshan 528200, China;
2. Institute of Animal Husbandry and Veterinary Science, Zhejiang Academy of Agricultural Sciences, Hangzhou 310021, China;
3. Taizhou Animal Disease Prevention and Control Center, Taizhou 317700, China

Received:2022-06-27 Online:2023-03-23 Published:2023-03-21

摘要/Abstract

摘要： 旨在调查胸膜肺炎放线杆菌流行株的血清型及Apx毒素基因携带情况，采用PCR方法对APP分离株进行血清型鉴定和Apx毒素基因检测。结果表明，64个分离株分别被鉴定为2、5、7、8、12、15、19共7个血清型。7个血清型中发现12种不同的Apx毒力因子模式。毒力因子分析结果显示，同一血清型的不同菌株携带的Apx毒素基因存在差异，不同血清型的菌株也可能会携带相同模式的Apx毒素基因。本研究表明，血清8型胸膜肺炎放线杆菌是浙江省的优势血清型，血清19型菌株在国内首次被检测到。血清型与所携带的Apx毒素基因之间不存在必然联系。

关键词: 胸膜肺炎放线杆菌, 血清分型, Apx, 毒力因子, PCR

Abstract: The study aimed to investigate the serotypes and Apx toxin genes of Actinobacillus pleuropneumoniae clinical isolates. PCR was used to identify the serotypes and detect the Apx toxin genes of APP isolates. The results showed that 64 isolates were identified as serotypes 2, 5, 7, 8, 12, 15, 19 respectively. A total of 12 different Apx virulence factor patterns were found in 7 serotypes. The results of virulence factor analysis showed that different strains of the same serotype carried different Apx toxin genes, and the strains of different serotypes may also carry the same pattern of Apx toxin genes. A. pleuropneumoniae serotype 8 was the dominant serotype in Zhejiang Province, and serotype 19 was detected for the first time in China. There is no correlation between the Apx toxin gene and the serotype.

Key words: Actinobacillus pleuropneumoniae, serotyping, Apx, virulence factor, PCR

中图分类号:

S852.619

卢碧凯, 袁秀芳, 徐丽华, 余斌, 苏菲, 叶十一, 陈怡洁, 蒋利明, 张晖, 李军星. 胸膜肺炎放线杆菌分离株血清型的PCR鉴定和Apx毒素基因检测[J]. 畜牧兽医学报, 2023, 54(3): 1341-1346.

LU Bikai, YUAN Xiufang, XU Lihua, YU Bin, SU Fei, YE Shiyi, CHEN Yijie, JIANG Liming, ZHANG Hui, LI Junxing. Molecular Serotyping and Apx Gene Profile Analysis of Actinobacillus pleuropneumoniae Isolates[J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(3): 1341-1346.

参考文献 21

[1]	SASSU E L, BOSSÉ J T, TOBIAS T J, et al. Update on Actinobacillus pleuropneumoniae-knowledge, gaps and challenges[J]. Transbound Emerg Dis, 2018, 65(Suppl 1):72-90.
[2]	PERRY M B, ALTMAN E, BRISSON J R, et al. Structural characteristics of the antigenic capsular polysaccharides and lipopolysaccharides involved in the serological classification of Actinobacillus (Haemophilus) pleuropneumoniae strains[J]. Serodiag Immunother Infect Dis, 1990, 4(4):299-308.
[3]	BLACKALL P J, KLAASEN H L B M, VAN DEN BOSCH H, et al. Proposal of a new serovar of Actinobacillus pleuropneumoniae:serovar 15[J]. Vet Microbiol, 2002, 84(1-2):47-52.
[4]	SÁRKÖZI R, MAKRAI L, FODOR L. Identification of a proposed new serovar of Actinobacillus pleuropneumoniae:serovar 16[J]. Acta Vet Hung, 2015, 63(4):444-450.
[5]	BOSSÉ J T, LI Y W, SÁRKÖZI R, et al. A unique capsule locus in the newly designated Actinobacillus pleuropneumoniae serovar 16 and development of a diagnostic PCR assay[J]. J Clin Microbiol, 2017, 55(3):902-907.
[6]	BOSSÉ J T, LI Y W, SÁRKÖZI R, et al. Proposal of serovars 17 and 18 of Actinobacillus pleuropneumoniae based on serological and genotypic analysis[J]. Vet Microbiol, 2018, 217:1-6.
[7]	STRINGER O W, BOSSÉ J T, LACOUTURE S, et al. Proposal of Actinobacillus pleuropneumoniae serovar 19, and reformulation of previous multiplex PCRs for capsule-specific typing of all known serovars[J]. Vet Microbiol, 2021, 255:109021.
[8]	刁有祥,丁家波,姜世金,等.地高辛标记探针检测猪胸膜肺炎放线杆菌的研究与应用[J].畜牧兽医学报, 2005, 36(6):585-589.DIAO Y X, DING J B, JIANG S J, et al. Study on Digoxigenin-labeled nucleic acid probe for detection of Actinobacillus pleuropneumoniae[J]. Acta Veterinaria et Zootechnica Sinica, 2005, 36(6):585-589.(in Chinese)
[9]	宣春玲,王贵平,李春玲,等.胸膜肺炎放线杆菌溶血外毒素研究进展[J].动物医学进展, 2009, 30(6):82-85.XUAN C L, WANG G P, LI C L, et al. Advance in apx toxins[J]. Progress in Veterinary Medicine, 2009, 30(6):82-85.(in Chinese)
[10]	KAMP E M, VERMEULEN T M, SMITS M A, et al. Production of apx toxins by field strains of Actinobacillus pleuropneumoniae and Actinobacillus suis[J]. Infect Immun, 1994, 62(9):4063-4065.
[11]	CHIERS K, DE WAELE T, PASMANS F, et al. Virulence factors of Actinobacillus pleuropneumoniae involved in colonization, persistence and induction of lesions in its porcine host[J]. Vet Res, 2010, 41(5):65.
[12]	GRAM T, AHRENS P. Improved diagnostic PCR assay for Actinobacillus pleuropneumoniae based on the nucleotide sequence of an outer membrane lipoprotein[J]. J Clin Microbiol, 1998, 36(2):443-448.
[13]	熊如月.猪传染性胸膜肺炎亚单位-灭活菌苗抗体检测方法建立及免疫抗体消长规律测定[D].武汉:华中农业大学, 2020.XIONG R Y. Establishment of antibody detection methods and determination of the antibody growth and decline law induced by porcine contagious pleuropneumonia bacterin-toxiod vaccine[D]. Wuhan:Huazhong Agricultural University, 2020.(in Chinese)
[14]	STRINGER O W, BOSSÉ J T, LACOUTURE S, et al. Proposal of Actinobacillus pleuropneumoniae serovar 19, and reformulation of previous multiplex PCRs for capsule-specific typing of all known serovars[J]. Vet Microbiol, 2021, 255:109021.
[15]	STHITMATEE N, SIRINARUMITR T, MAKONKEWKEYOON L, et al. Identification of the Actinobacillus pleuropneumoniae serotype using PCR based-apx genes[J]. Mol Cell Probes, 2003, 17(6):301-305.
[16]	SCHUWERK L, HOELTIG D, WALDMANN K H, et al. Sero-and apx-typing of German Actinobacillus pleuropneumoniae field isolates from 2010 to 2019 reveals a predominance of serovar 2 with regular apx-profile[J]. Vet Res, 2021, 52(1):10.
[17]	GOTTSCHALK M, LACOUTURE S. Canada:distribution of Streptococcus suis(from 2012 to 2014) and Actinobacillus pleuropneumoniae(from 2011 to 2014) serotypes isolated from diseased pigs[J]. Can Vet J, 2015, 56(10):1093-1094.
[18]	KIM B, HUR J, LEE J Y, et al. Molecular serotyping and antimicrobial resistance profiles of Actinobacillus pleuropneumoniae isolated from pigs in South Korea[J]. Vet Q, 2016, 36(3):137-144.
[19]	刘建杰,陈焕春,何启盖,等.猪传染性胸膜肺炎新型亚单位菌苗对小鼠的效力研究[J].畜牧兽医学报, 2005, 36(2):177-180.LIU J J, CHEN H C, HE Q G, et al. Protective efficacy of a genetic subunit bacterin against Actinobacillus pleuropneumoniae in mice[J]. Acta Veterinaria et Zootechnica Sinica, 2005, 36(2):177-180.(in Chinese)
[20]	夏新萌,连伟民,李成蒙.我国部分地区猪传染性胸膜肺炎放线杆菌的血清型流行病学调查[J].中国猪业, 2015, 10(12):50-53.XIA X M, LIAN W M, LI C M. Porcine infectious pleuropneumonia in some areas of Chinese serotype epidemiological investigation of actinobacter[J]. China Swine Industry, 2015, 10(12):50-53.(in Chinese)
[21]	廖永洪.猪胸膜肺炎放线杆菌免疫蛋白质组学及亚单位疫苗的研究[D].武汉:华中农业大学, 2010.LIAO Y H. Immunoproteomic analysis and study on subunit vaccine of Actinobacillus Pleuroneumoniae[D]. Wuhan:Huazhong Agricultural University, 2010.(in Chinese)

胸膜肺炎放线杆菌分离株血清型的PCR鉴定和Apx毒素基因检测

Molecular Serotyping and Apx Gene Profile Analysis of Actinobacillus pleuropneumoniae Isolates

RichHTML

PDF

可视化

摘要/Abstract

引用本文

使用本文

参考文献 21

相关文章 15

编辑推荐

Metrics

本文评价

[1]	杨恒, 李占鸿, 宋子昂, 高林, 李卓然, 廖德芳, 肖雷, 李华春. 帕利亚姆病毒实时荧光定量RT-PCR检测方法的建立与应用[J]. 畜牧兽医学报, 2024, 55(1): 395-400.
[2]	周建浩, 王东方, 刘影, 王淑娟, 马震原, 谢彩华, 赵雪丽, 杨海波, 冯桂丹, 康台生, 胡煜锋, 李博文, 闫若潜. 猪流行性腹泻病毒微滴式数字PCR定量检测方法的建立及初步应用[J]. 畜牧兽医学报, 2024, 55(1): 413-418.
[3]	姜玲玲, 牛小霞, 刘强, 张刚, 王璞, 李勇. 宁夏地区肉牛腹泻相关病毒感染状况的分析[J]. 畜牧兽医学报, 2023, 54(9): 3863-3871.
[4]	陈曦, 王一, 王佳丽, 杨新, 宋军科, 赵光辉. 毒害艾美耳球虫和产气荚膜梭菌双重PCR检测方法的建立[J]. 畜牧兽医学报, 2023, 54(9): 3985-3990.
[5]	杨富升, 古小彬. 近十年PCR技术在寄生虫病诊断中的应用[J]. 畜牧兽医学报, 2023, 54(8): 3183-3194.
[6]	邱英武, 常昊, 彭杰, 易和友, 王秋梅, 郭彦辰, 吴倩雯, 曹雪珍, 林丽苗, 李薇, 周庆丰, 张桂红, 李群辉, 龚浪. 一种快速检测非洲猪瘟病毒I177L基因缺失毒株的荧光定量PCR方法[J]. 畜牧兽医学报, 2023, 54(6): 2521-2527.
[7]	王宏宇, 朱寅初, 云涛, 张存, 鲍恩东. 鹅星状病毒基因Ⅰ型和Ⅱ型双重荧光定量RT-PCR检测方法的建立与应用[J]. 畜牧兽医学报, 2023, 54(4): 1616-1623.
[8]	何姝凡, 邹沅彤, 黄智兰, 李倩, 降措翁西, 岳华, 汤承, 刘杰. 牛腺病毒3型感染状况及其fiber轴区基因检测[J]. 畜牧兽医学报, 2023, 54(3): 1333-1340.
[9]	翟刚, 顾文源, 刘涛, 刘莹, 张帅, 范京惠, 左玉柱. 猪流行性腹泻病毒TaqMan检测方法的建立及基于S基因的遗传变异分析[J]. 畜牧兽医学报, 2023, 54(2): 847-854.
[10]	支岩, 梅晨, 刘珍邑, 乌云格日乐, 王宏俊, 胡格. 副鸡禽杆菌毒力因子研究进展[J]. 畜牧兽医学报, 2023, 54(12): 4934-4942.
[11]	肖洁, 罗跃军, 陈浩, 蒲家艳, 何维, 熊常明, 郝哥, 任永军, 杨光友. 兔斯氏艾美耳球虫、肠艾美耳球虫和大型艾美耳球虫PCR检测靶标的筛选与单一、多重直接PCR检测方法的建立[J]. 畜牧兽医学报, 2023, 54(10): 4327-4337.
[12]	邹宏, 夏应菊, 李玲, 徐璐, 赵俊杰, 王团结, 张乾义, 宋振辉. 猪瘟病毒和牛病毒性腹泻病毒双重荧光定量PCR检测方法的建立和初步应用[J]. 畜牧兽医学报, 2023, 54(10): 4422-4427.
[13]	常昊, 邱英武, 彭杰, 高琦, 宋泽布, 陈洋, 李薇, 林丽苗, 曹雪珍, 周庆丰, 张桂红, 李群辉, 郑泽中. 非洲猪瘟病毒基因Ⅰ型的双重实时荧光定量PCR检测方法建立[J]. 畜牧兽医学报, 2023, 54(10): 4428-4432.
[14]	龚三你, 蒋旭东, 马瑶, 卢建远, 字向东. 牦牛FHL2基因的克隆、组织表达与蛋白定位分析[J]. 畜牧兽医学报, 2023, 54(1): 146-156.
[15]	张凯川, 王晋宇, 李守军, 贾坤. 广东省羊源肺炎克雷伯菌遗传进化与毒力基因及耐药性分析[J]. 畜牧兽医学报, 2023, 54(1): 328-337.