摘要
为了解猪链球菌血清9型菌株KQ-1株、KQ-2株的生物特性,对其进行了菌种鉴定、生长曲线绘制、毒力因子鉴定和小鼠致病性试验。首先采用革兰氏染色、PCR鉴定的方法对2株菌进行菌种鉴定,然后利用猪链球菌的主要毒力基因mrp、epf、sly、gadph、orf2、fbps、89k的引物对2株菌的毒力基因进行鉴定;将2株菌在TSB培养基(含10%新生牛血清)中进行培养,绘制其生长曲线,初步了解这2株菌的生长特性;选取45只健康的昆明鼠注射2株菌的菌液,进行致病性试验,了解其致病性。结果显示,KQ-1株、KQ-2株革兰氏染色呈阳性,PCR鉴定结果为猪链球菌血清9型;2株菌均在37℃培养6 h时菌液密度达到最高值。对2株菌毒力基因的鉴定结果如下:KQ-1株的基因型为mrp+/epf-/sly+/gadph+/orf2-/fbps-/89k-,KQ-2株的基因型为mrp-/epf-/sly-/gadph+/orf2+/fbps-/89k-;对2株菌的小鼠致病性试验表明,2株菌均为强毒株。
In order to understand the biological characteristics of Streptococcus suis serotype 9 strains(KQ-1 and KQ-2),the identification of strains and their virulence genes,drawing of growth curves,mouse’s pathogenicity test were conducted.Those two strains were identified by Gram staining and PCR,then the primers of the main virulence genes(mrp,epf,sly,gadph,orf2,fbps,89k)of S.suis were used to identify the virulence genes of the two strains.The two strains were cultured in TSB medium(containing 10%newborn calf serum),and their growth curves were drawn to get a preliminary understanding of the growth characteristics of the two strains.45 healthy Kunming rats were injected with two strains of bacteria for the pathogenicity test to understand their pathogenicity.The results showed that the Gram staining results of KQ-1 strain and KQ-2 strain were positive,they were identified as S.suis serotype 9 strains by PCR identification.The bacterial liquid density of the two strains both reached the peak at 6 h when cultured at 37℃.The results of virulence factors of the two strains showed that the genotype of KQ-1 strain was mrp+/epf-/sly+/gadph+/orf2-/fbps-/89k-,while that of KQ-2 strain was mrp-/epf-/sly-/gadph+/orf2+/fbps-/89k-.The results of pathogenicity test on mice showed that the two strains were highly virulent.
作者
王召贺
陈波
徐高原
张华伟
郝根喜
潘建刚
周明光
WANG Zhao-he;CHEN Bo;XU Gao-yuan(Wuhan Keqian Biology Co.,Ltd.,Wuhan,Hubei 430000)
出处
《安徽农业科学》
CAS
2021年第10期92-97,121,共7页
Journal of Anhui Agricultural Sciences
关键词
猪链球菌
血清9型
毒力基因
致病性试验
Streptococcus suis
Serotype 9
Virulence genes
Pathogenicity test
