Identification of pathogenicity, investigation of virulent gene distribution and development of a virulent strain-specific detection PCR method for Vibrio harveyi isolated from Hainan Province and Guangdong Province, China
Identification of pathogenicity, investigation of virulent gene distribution and development of a virulent strain-specific detection PCR method for Vibrio harveyi isolated from Hainan Province and Guangdong Province, China
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海南省和广东省哈维氏弧菌致病性鉴定、毒力基因分布调查及毒力菌株特异性检测PCR方法的建立
DOI：
10.1016/j.aquaculture.2016.10.015
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发表时间：
2017-02
期刊：
影响因子：
4.5
通讯作者：
Zhou Yongcan
中科院分区：
文献类型：
--
作者：
Xu Xi;ong;Liu Kaifang;Wang Shifeng;Guo Weiliang;Xie Zhenyu;Zhou Yongcan
从对虾中分离到46株哈维氏弧菌，2011年至2013年期间，中国海南省和广东省沿海地区的笛鲷和其他海水养殖鱼类。通过人工感染107个菌落形成单位（CFU）mL− 1的健康的斜带石斑鱼，18个菌株被确定为致病性。死亡发生在注射后2至6小时内。肠细菌重复基因间共识-聚合酶链反应（ERIC-PCR）的46株菌株的基因型检测显示了类似的基因型谱，称为ERIC-1的配置文件，所有18个致病菌株。这一发现表明，致病性V.中国南方的harveyistrains具有相似的遗传背景，可能是该地区的代表性致病菌株。对所有46个菌株进行了筛选，以确定是否存在典型的毒力基因。harveyi，动物源性弧菌，如V.霍乱弧菌、副溶血性弧菌和霍乱弧菌。vulnificusand of the aquatic pathogenV.鳗属用特异性引物PCR扩增出5个哈维氏分支的典型毒力基因：luxR、toxRvh、chiA、丝氨酸蛋白酶和vhh。非致病性菌株仅携带1 ~ 4个基因，表明这5个基因可能是ERIC-1菌株的主要毒力基因。根据18株病原菌ERIC-PCR扩增产物的序列，设计了菌株特异性PCR引物。种特异性引物具有较高的特异性和敏感性。这项研究表明，细菌是非常重要的海水养殖可以特异性检测使用ERIC-PCR指纹扩增。
A collection of 46Vibrio harveyistrains were isolated fromEpinephelusspp.,Lutjanus erythopterus, and other maricultured fish in coastal areas of Hainan Province and Guangdong Province, China, between 2011 and 2013. Eighteen strains were determined to be pathogenic via artificial infection of healthyEpinephelus coioidesat 107colony-forming units (CFU) mL− 1. Mortality occurred within 2 to 6 h after injection. Genotypic assays of the 46 strains by enterobacterial repetitive intergenic consensus-polymerase chain reaction (ERIC-PCR) revealed a similar genotype profile, referred to as the ERIC-1 profile, for all 18 pathogenic strains. This finding indicates that pathogenicV. harveyistrains in south China have similar genetic backgrounds and might be representative pathogenic strains of this region. All 46 strains were screened for the presence of virulence genes typical ofV. harveyi, of zoonoticVibriospecies such asV. cholerae,V. parahaemolyticus, andV. vulnificusand of the aquatic pathogenV. anguillarum. Virulence genes were amplified by PCR using specific primers, and five typical virulence genes of the Harveyi clade,luxR,toxRvh,chiA,serine proteaseandvhh, were detected in all pathogenic isolates. Non-pathogenic strains carried only 1 to 4 of these genes, indicating that these five genes might be the main virulence genes of ERIC-1 strains. Strain-specific PCR primers were designed based on the sequences of distinct ERIC-PCR bands for the 18 pathogenic strains. Species-specific primers exhibited high specificity and sensitivity. This study demonstrates that bacteria that are highly important to mariculture could be specifically detected using ERIC-PCR fingerprint-based amplification.
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DOI：
10.1016/j.scitotenv.2009.12.019
发表时间：
2010-02
期刊：
The Science of the total environment
影响因子：
--
作者：
W. Yuan;T. Chai;Z. Miao
通讯作者：
W. Yuan;T. Chai;Z. Miao
影响因子：
1.2
作者：
Conejero, MJU;Hedreyda, CT
通讯作者：
Hedreyda, CT
影响因子：
2.2
作者：
Y. Ye;Qingping Wu;Yanhong Zhou;Xiaohui Dong;Jumei Zhang
通讯作者：
Y. Ye;Qingping Wu;Yanhong Zhou;Xiaohui Dong;Jumei Zhang
影响因子：
2.4
作者：
Alippi, AM;López, AC;Aguilar, OM
通讯作者：
Aguilar, OM
影响因子：
2.6
作者：
Bliss, CI
通讯作者：
Bliss, CI