辽宁省猪链球菌流行株主要血清型毒力因子检测及其致病性
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摘要
为了解辽宁省猪链球菌流行菌株主要血清型毒力因子分布特征及致病性,依据猪链球菌属保守基因及血清型特异性基因,从辽宁地区采集猪链球菌疑似病例肺脏、肝脾、脑、关节液等样品进行猪链球菌及其血清类型鉴定;应用猪链球菌6种主要毒力因子特异性基因扩增检测方法,检测所分离到的不同血清类型猪链球菌的毒力因子分布情况,并应用小鼠攻毒试验和病理学技术对其致病性进行观察研究。结果显示:从辽宁地区采集的72个样品中共分离到23株猪链球菌,主要血清型有1,2,7型,其中SS1型4株,SS2型2株,SS7型5株。其余12株未确定血清型。SS1、SS2、SS7型阳性率分别为17.39%,8.69%,30.43%。毒力因子检测结果显示,上述6种毒力因子检出率分别为100%,66.6%,25%,16.7%,75.0%,58.3%,其中SS2均具备6种毒力因子,SS1、SS7和血清型阴性株猪链球菌均具有部分毒力因子,与SS2的差异毒力因子为epf。SS2可引起小鼠的急性败血症以及脑膜炎,SS1、SS7均可引起小鼠发病,但各器官的损伤则较轻。SS1、SS2、SS7脑内接种均可引起小鼠脑膜炎,炎症反应SS2最重,SS1次之,SS7最轻。
In order to understand the distribution characteristics and pathogenicity of the virulence factors of main serotype Streptococcus suis in Liaoning Province,according to the conservative genes and serotype-specific genes of Streptococcus,the suspected cases of Streptococcus suis from the lungs,liver,spleen,and brain,synovial fluid and other samples were used to isolate and identify Streptococcus suis and their serotypes.Six virulence factor-specific gene amplification assays for Streptococcus suis were used to detect the distribution of virulence factors in the different serotypes of Streptococcus suis,and the application of mouse challenge test and pathological techniques to observe its pathogenicity.The results showed that 23 strains of Streptococcus suis were isolated from 72 samples collected in Liaoning.The main serotypes were type 1,2 and 7 among which were SS1 type 4,SS2 type 2 and SS7 type 5 strains,respectively.The remaining 12 strains were not serotyped.The positive rates of SS1,2,7 were 17.39%,8.69% and 30.43%,respectively.The virulence factor test results showed that the detection rates of the above six virulence factors were 100%,66.6%,25%,16.7%,75%and 58.3%,respectively,of which SS2 had 6 virulence factors,SS1,SS7 and the serotype-negative strain of Streptococcus suis had partial virulence factors,and the differential virulence factor of SS2 was epf;SS2 could cause acute septicemia and meningitis in mice.SS1 and SS7 could cause disease in mice,but the injury of each organ was slight.Intracerebral injection of SS1,SS2,and SS7 could cause meningitis in mice,and inflammatory response caused by SS2 was the most important,followed by SS1,SS7 lightest.
In order to understand the distribution characteristics and pathogenicity of the virulence factors of main serotype Streptococcus suis in Liaoning Province,according to the conservative genes and serotype-specific genes of Streptococcus,the suspected cases of Streptococcus suis from the lungs,liver,spleen,and brain,synovial fluid and other samples were used to isolate and identify Streptococcus suis and their serotypes.Six virulence factor-specific gene amplification assays for Streptococcus suis were used to detect the distribution of virulence factors in the different serotypes of Streptococcus suis,and the application of mouse challenge test and pathological techniques to observe its pathogenicity.The results showed that 23 strains of Streptococcus suis were isolated from 72 samples collected in Liaoning.The main serotypes were type 1,2 and 7 among which were SS1 type 4,SS2 type 2 and SS7 type 5 strains,respectively.The remaining 12 strains were not serotyped.The positive rates of SS1,2,7 were 17.39%,8.69% and 30.43%,respectively.The virulence factor test results showed that the detection rates of the above six virulence factors were 100%,66.6%,25%,16.7%,75%and 58.3%,respectively,of which SS2 had 6 virulence factors,SS1,SS7 and the serotype-negative strain of Streptococcus suis had partial virulence factors,and the differential virulence factor of SS2 was epf;SS2 could cause acute septicemia and meningitis in mice.SS1 and SS7 could cause disease in mice,but the injury of each organ was slight.Intracerebral injection of SS1,SS2,and SS7 could cause meningitis in mice,and inflammatory response caused by SS2 was the most important,followed by SS1,SS7 lightest.
引文
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[23] SUN Y,LIU H,DU R,et al.Characteristic comparison of meningitis and non-meningitis of Streptococcus suis in an experimentally infected porcine[J].Inflammation,2018,41(2):368-377.
[2] WERTHEIM H F,NGHIA H D,TAYLOR A W,et al.Streptococcus suis:an emerging human pathogen[J].Clin Infect Dis,2009,48(5):617-625.
[3] STAATS J J,FEDER I,OKWUMABUA O,et al.Streptococcus suis:past and present[J].Vet Res Commun,1997,21(6):381-407.
[4] 邹君,王楷成,田莉莉,等.链球菌猪主要致病种和型多重PCR检测方法的建立[J].中国兽医科学,2011,41(7):701-706.
[5] 李干武,姚火春,陆承平.在猪链球菌2型江苏分离株中发现新的毒力相关基因[J].农业生物技术学报,2003,11(3):295-298.
[6] 方勤美,黄绍谦,俞伏松,等.猪链球菌2型福建分离株的主要毒力基因分析[J].中国人兽共患病学报,2010,26(1):65-68.
[7] 郑升博,华修国,陆伟芳,等.多重PCR方法检测猪链球菌主要致病血清型及其毒力因子[J].上海交通大学学报,2010,28(1):53-58.
[8] 王巍.四川地区猪链球菌流行调查及其病原特性研究[D].四川雅安:四川农业大学,2015.
[9] 马增军.河北省猪链球菌病病原学调查与分析[D].北京:中国农业大学,2013.
[10] 王娟.2014—2015年广东部分地区猪链球菌流行病学调查[D].江苏南京:南京农业大学,2015.
[11] 王淑杰,雷连成,徐敏,等.东北地区猪群链球菌分离鉴定及流行病学调查分析[J].中国兽医学报,2009,29(7):877-881.
[12] 刘耀川,宋淑英,张泽辉,等.猪链球菌毒力因子简介[J].湖北畜牧兽医,2015,36(3):7-8.
[13] 赵冉,孙建和,陆承平.猪链球菌国内分离株毒力因子的分布特征[J].上海交通大学学报,2006,24(6):495-498.
[14] VECHT U,WISSELINK H J,JELLEMA M L,et al.Identification of two proteins associated with virulence of the Streptococcus suis type 2[J].Infect Immun,1992,59:3156-3162.
[15] JEAN-PHILIPPE AUGE R,NAHUEL FITTIPALD I,MARIE-ODIL E,et al.Virulence studies of different sequence types and geographical origins of Streptococcus suis serotype 2 in a mouse model of infection[J].Pathogens,2016,5(3).E48.doi:10.3390/pathogens5030048.
[16] 胡丹,石洁,朱静,等.猪链球菌2型BALB/c小鼠动物感染模型的建立[J].中国病原生物学杂志,2012,7(8):561-564.
[17] WILLIAMS A E,BLAKEMORE W F,ALEXANDER T J.A murine model of Streptococcus suis type 2 meningitis in the pig[J].Res Vet Sci,1988,45(3):394-9.
[18] KATAOKA Y,HARITANT M,MORI M,et al.Experimental infections of mice and pigs with Streptococcus suis type 2[J].J Vet Med Sci,1991,53(6):1043-1049.
[19] 王继春,何孔旺,何家惠,等.2型猪链球菌动物致病性试验[[J].中国兽医科技,2001,31(12):23-25.
[20] DE GREEFF A,VAN SELM S,BUYS H,et al.Pneumococcal colonization and invasive disease studied in a porcine model[J].BMC Microbiol,2016,16:102.
[21] BI Y,LI J,YANG L,et al.Assessment of the pathogenesis of Streptococcus suis type 2 infection in piglets for understanding streptococcal toxic shock-like syndrome,meningitis,and sequelae[J].Vet Microbiol,2014,173:299-309.
[22] VECHT U,STOCKHOFE-ZURWIEDEN N,TETENBURG B J,et al.Virulence of Streptococcus suis type 2 for mice and pigs appeared host-specific[J].Vet Microbiol,1997,58(1):53-60.
[23] SUN Y,LIU H,DU R,et al.Characteristic comparison of meningitis and non-meningitis of Streptococcus suis in an experimentally infected porcine[J].Inflammation,2018,41(2):368-377.