山羊源产单核细胞李斯特菌的分离与鉴定
|
摘要
从贵州省盘县某养羊场病例组织样本中分离出1株疑似产单核细胞李斯特菌(Lm)。通过革兰染色、生化鉴定、PCR扩增hly基因、克隆、测序、序列分析及药敏试验等方法对可疑菌株进行分析鉴定。结果显示,该分离菌的培养特性、菌落形态、菌体形状特征、生理生化特征均与文献报道的产单核细胞李斯特菌相同,并从该分离菌株基因组中扩增到大小约850bp的Lm的特异性DNA片段,其序列与GenBank中其他Lm地方株核苷酸同源性在39.1%~99.7%之间,其中与从发病动物分离到的加拿大、瑞士参考株同源性最高,均达到99.7%,与其他途径分离到的参考菌株同源性都很低,分子水平上进一步证实分离菌株为产单核细胞李斯特菌,且从不同途径分离的地方株hly基因差异大。研究结果为本病临床防控与治疗提供理论依据。
AListeria monocytogenes(Lm)was isolated from the tissue samples of a infected case in a caprine farm in Pan county of Guizhou province.The isolated strain was analysed and identified by means of Gram staining,biochemical identification,PCR amplification of hly gene,cloning,sequencing,sequence analysis and drug susceptibility test.The results showed that a suspected Lm was isolated from a suspected caprine case,and its cultural characteristics,colony morphology,and physiological-biochemical characteristics were the same as the Lm in literature reports.A specific DNA fragment about 850 bp of Lm was amplified.The nucleotide homology is between 39.1%-99.7%compared with part of strains in GenBank,and is the higest with the strains from Canada and Switzerland(99.7%),very low with the other strains isolated from other ways.The result from the molecular level confirmed that the isolate is a Lm,and the genetic differences between the hly genes of local strains isolated from different ways were significant.The results can provide theoretical data for prevention and treatment of the disease.
AListeria monocytogenes(Lm)was isolated from the tissue samples of a infected case in a caprine farm in Pan county of Guizhou province.The isolated strain was analysed and identified by means of Gram staining,biochemical identification,PCR amplification of hly gene,cloning,sequencing,sequence analysis and drug susceptibility test.The results showed that a suspected Lm was isolated from a suspected caprine case,and its cultural characteristics,colony morphology,and physiological-biochemical characteristics were the same as the Lm in literature reports.A specific DNA fragment about 850 bp of Lm was amplified.The nucleotide homology is between 39.1%-99.7%compared with part of strains in GenBank,and is the higest with the strains from Canada and Switzerland(99.7%),very low with the other strains isolated from other ways.The result from the molecular level confirmed that the isolate is a Lm,and the genetic differences between the hly genes of local strains isolated from different ways were significant.The results can provide theoretical data for prevention and treatment of the disease.
引文
[1]陆承平.兽医微生物学[M].5版.北京:中国农业出版社,2013:179.
[2]李浩欣,段纲,项勋,等.仔猪李斯特杆菌的分离与鉴定[J].中国畜牧兽医,2015(7):1883-1889.
[3]杨建伟.单核细胞增生性李斯特菌最新研究进展[J].齐鲁医学杂志,2013(5):465-467.
[4]田静.熟肉制品中单增李斯特菌的风险评估及风险管理措施的研究[D].北京:中国疾病预防控制中心,2010.
[5]Takahashi T,Ochiai Y,Matsudate H,et al.Isolation of Listeria monocytogenes from the skin of slaughtered beef cattle[J].J Vet Med Sci,2007,69(10):1077-1079.
[6]Orsi R H,den Bakker H C,Wiedmann M.Listeria monocytogeneslineages:Genomics,evolution,ecology,and phenotypic characteristics[J].Int J Med Microbiol,2011,301(2):79-96.
[7]陈溥言.兽医传染病学[M].5版.北京:中国农业出版社,2006:162.
[8]崔焕忠,乔立桥,王义冲.单核细胞增生性李斯特菌的主要毒力因子及其致病机理[J].中国畜牧兽医,2010(1):128-133.
[9]Vanghele M,Ganea E.The role of bacterial molecular chaperones in pathogen survival within the host[J].Rom J Biochem,2010,47(87):100.
[10]王国梁,殷月兰,焦库华,等.绵羊李斯特菌病病原诊断及其生物学特性研究[J].中国人兽共患病学报,2013(7):639-645.
[11]贺春月,王艳,王天姝,等.中国部分食品来源单增李斯特菌中致病相关基因的分布研究[J].疾病监测,2013(5):384-388.
[12]董慧.产单核细胞李斯特菌hly和actA基因的原核表达及其产物的单克隆抗体研制[D].江苏扬州:扬州大学,2006.
[13]周晓辉,焦新安.产单核细胞李斯特菌的分子鉴定与亚分型研究[J].中国人兽共患病杂志,2003,19(5):44-47.
[14]周环.牛李斯特菌脑炎的流行、诊断与防治[J].现代畜牧科技,2015(2):126.
[15]Bhasin S K,Chaturvedi S,Sharma A K,et al.Knowledge amongst adult population regarding vectors of malaria in 21states of India[J].J Commun Dis,2001,33(4):286-296.
[16]蒋原.食源性病原微生物检测指南[M].北京:中国标准出版社,2010:113-115.
[17]Nightingale K K,Windham K,Wiedmam M.Evolution and molecular phylogeny of Listeria monocytogenes isolated from human and animal listeriosiscases and foods[J].J Bacteriol,2005,187(16):5537-5551.
[18]贾艳艳,何雷,郁川,等.豫西地区市售鸭翅中单增李斯特菌毒力基因的检测[J].中国畜牧兽医,2014(11):283-287.
[19]肖开提,冬不拉提,白军民,等.一起羊李氏杆菌病的诊断及病因分析[J].中国畜牧兽医,2007(11):117-118.
[20]尹录,杜艳芬,赫明雷,等.哈尔滨市鲜肉中单核细胞增生性李斯特菌的分离鉴定及耐药性分析[J].中国预防兽医学报,2009(12):929-932.
[21]王文燕.猪肉中单核细胞增生李斯特菌的分离与耐药性筛查[D].广东广州:华南理工大学,2014.
[2]李浩欣,段纲,项勋,等.仔猪李斯特杆菌的分离与鉴定[J].中国畜牧兽医,2015(7):1883-1889.
[3]杨建伟.单核细胞增生性李斯特菌最新研究进展[J].齐鲁医学杂志,2013(5):465-467.
[4]田静.熟肉制品中单增李斯特菌的风险评估及风险管理措施的研究[D].北京:中国疾病预防控制中心,2010.
[5]Takahashi T,Ochiai Y,Matsudate H,et al.Isolation of Listeria monocytogenes from the skin of slaughtered beef cattle[J].J Vet Med Sci,2007,69(10):1077-1079.
[6]Orsi R H,den Bakker H C,Wiedmann M.Listeria monocytogeneslineages:Genomics,evolution,ecology,and phenotypic characteristics[J].Int J Med Microbiol,2011,301(2):79-96.
[7]陈溥言.兽医传染病学[M].5版.北京:中国农业出版社,2006:162.
[8]崔焕忠,乔立桥,王义冲.单核细胞增生性李斯特菌的主要毒力因子及其致病机理[J].中国畜牧兽医,2010(1):128-133.
[9]Vanghele M,Ganea E.The role of bacterial molecular chaperones in pathogen survival within the host[J].Rom J Biochem,2010,47(87):100.
[10]王国梁,殷月兰,焦库华,等.绵羊李斯特菌病病原诊断及其生物学特性研究[J].中国人兽共患病学报,2013(7):639-645.
[11]贺春月,王艳,王天姝,等.中国部分食品来源单增李斯特菌中致病相关基因的分布研究[J].疾病监测,2013(5):384-388.
[12]董慧.产单核细胞李斯特菌hly和actA基因的原核表达及其产物的单克隆抗体研制[D].江苏扬州:扬州大学,2006.
[13]周晓辉,焦新安.产单核细胞李斯特菌的分子鉴定与亚分型研究[J].中国人兽共患病杂志,2003,19(5):44-47.
[14]周环.牛李斯特菌脑炎的流行、诊断与防治[J].现代畜牧科技,2015(2):126.
[15]Bhasin S K,Chaturvedi S,Sharma A K,et al.Knowledge amongst adult population regarding vectors of malaria in 21states of India[J].J Commun Dis,2001,33(4):286-296.
[16]蒋原.食源性病原微生物检测指南[M].北京:中国标准出版社,2010:113-115.
[17]Nightingale K K,Windham K,Wiedmam M.Evolution and molecular phylogeny of Listeria monocytogenes isolated from human and animal listeriosiscases and foods[J].J Bacteriol,2005,187(16):5537-5551.
[18]贾艳艳,何雷,郁川,等.豫西地区市售鸭翅中单增李斯特菌毒力基因的检测[J].中国畜牧兽医,2014(11):283-287.
[19]肖开提,冬不拉提,白军民,等.一起羊李氏杆菌病的诊断及病因分析[J].中国畜牧兽医,2007(11):117-118.
[20]尹录,杜艳芬,赫明雷,等.哈尔滨市鲜肉中单核细胞增生性李斯特菌的分离鉴定及耐药性分析[J].中国预防兽医学报,2009(12):929-932.
[21]王文燕.猪肉中单核细胞增生李斯特菌的分离与耐药性筛查[D].广东广州:华南理工大学,2014.