采集新疆奶牛隐性乳腺炎乳样,对分离鉴定的致病性金黄色葡萄球菌,提取分离菌株的基因组DNA,参考GenBank中发表的ClfA(Clumping Factor A of Staphylococcus aureus)基因序列,设计特异性引物经PCR扩增得到1 560bp的ClfA基因。对新疆3个...采集新疆奶牛隐性乳腺炎乳样,对分离鉴定的致病性金黄色葡萄球菌,提取分离菌株的基因组DNA,参考GenBank中发表的ClfA(Clumping Factor A of Staphylococcus aureus)基因序列,设计特异性引物经PCR扩增得到1 560bp的ClfA基因。对新疆3个地区分离的9株金黄色葡萄球菌分别进行ClfA基因的克隆和序列分析,对ClfA蛋白的氨基酸序列分析结果表明,新疆3个地区ClfA蛋白氨基酸的同源性为91.5%～100%,与国外其他分离菌株的同源性为91.9%～100%,差异性较小,该结果表明ClfA基因在该菌进化和流行过程中是保守的。展开更多
Objective:To analysis and identify a bacterium strain isolated from laboratory breeding mouse far away from a hospital.Methods:Phenotype of the isolate was investigated by conventional microbiological methods,includin...Objective:To analysis and identify a bacterium strain isolated from laboratory breeding mouse far away from a hospital.Methods:Phenotype of the isolate was investigated by conventional microbiological methods,including Gram-staining,colony morphology,tests for haemolysis, catalase,coagulase,and antimicrobial susceptibility test.The meek and 16S rRNA genes were amplified by the polymerase chain reaction(PCR) and sequenced.The base sequence of the PCR product was compared with known 16S rRNA gene sequences in the CenBank database by phylogenetic analysis and multiple sequence alignment.Results:The isolate in this study was a gram positive,coagulase negative,and catalase positive coccus.The isolate was resistant to oxacillin,methicillin,penicillin,ampicillin,cefazolin,cipr of loxacin erythromycin,et al.PCR results indicated that the isolate was meek gene positive and its 16S rRNA was 1465 bp.Phylogenetic analysis of the resultant 16S rRNA indicated the isolate belonged to genus Saphylococcus,and multiple sequence alignment showed that the isolate was Saphylococcus haemolyticus with only one base difference from the corresponding 16S rRNA deposited in the CenBank.Conclusions: 16S rRNA gene sequencing is a suitable technique for non-specialist researchers.Laboratory animals are possible sources of lethal pathogens,and researchers must adapt protective measures when they manipulate animals.展开更多
【目的】探索宁夏干旱荒漠区苦豆子内生放线菌的多样性及区系组成,为苦豆子内生放线菌的合理开发和利用提供理论依据。【方法】从宁夏白芨滩自然保护区不同植被和土壤类型的6个样区采集健康苦豆子样品30份,采用组织匀浆法从苦豆子植株...【目的】探索宁夏干旱荒漠区苦豆子内生放线菌的多样性及区系组成,为苦豆子内生放线菌的合理开发和利用提供理论依据。【方法】从宁夏白芨滩自然保护区不同植被和土壤类型的6个样区采集健康苦豆子样品30份,采用组织匀浆法从苦豆子植株的根部、茎部、叶部和种子中分离内生放线菌,根据培养性状、菌落、孢子等的形态特征和16S r RNA基因序列分析对分离菌株进行鉴定;根据苦豆子内生放线菌的相对频率、物种多样性指数、丰富度指数和相似性系数分析其区系组成特点。【结果】从30份苦豆子样品中,共分离得到内生放线菌111株,这些菌株分属于链霉菌属(Streptomyces)、拟诺卡氏菌属(Nocardiopsis)、普劳斯氏菌属(Prauserella)、Actinophytocola、微杆菌属(Microbacterium)、束丝放线菌属(Actinosynnema)、嗜热油菌属(Thermoleophilum)、糖霉菌属(Glycomyces)和糖丝菌属(Saccharothrix)9个属,其中链霉菌属(Streptomyces)和拟诺卡氏菌属(Nocardiopsis)为优势属。苦豆子内生放线菌的分布具有一定的组织特异性,根和种子中的内生放线菌数量高于茎和叶。植被类型中,沙生植被草原(样区Ⅳ)分离的苦豆子内生放线菌物种多样性指数最高,而荒漠草原(样区Ⅴ)最低。荒漠草原(样区Ⅴ)和林地(样区Ⅵ)内生放线菌群落有密切的相似性。【结论】苦豆子体内含有丰富的内生放线菌资源,其内生放线菌具有很高的宿主特异性,而且其分布受生境影响。展开更多
【目的】了解新疆地区牛病毒性腹泻（BVD）的流行状况,【方法】采用RT-PCR方法对从新疆16个不同地区采集的样品进行BVD病毒5＇-UTR扩增,对扩增出的序列进行测序,构建系统遗传进化树和遗传变异分析。【结果】566份样品中,扩增出21份阳性...【目的】了解新疆地区牛病毒性腹泻（BVD）的流行状况,【方法】采用RT-PCR方法对从新疆16个不同地区采集的样品进行BVD病毒5＇-UTR扩增,对扩增出的序列进行测序,构建系统遗传进化树和遗传变异分析。【结果】566份样品中,扩增出21份阳性样品,平均阳性率为3.71℅。通过对分离株序列的同源性比对分析,17株为BVDVI型,4株为BVDVⅡ型。经对PCR扩增阳性样品进行细胞传代,有7株为致细胞病变型（CP）,14株为非致细胞病变型（NCP）毒株,所测得的病毒滴度为1.04×103~6.02×108TCID50/0.1 m L,21株分离病毒对BVDV阳性血清的中和效价为1：21~1：25。【结论】BVD在新疆地区均不同程度的发生和流行。研究的成果,为BVD的有效防控提供了技术资料。展开更多
文摘采集新疆奶牛隐性乳腺炎乳样,对分离鉴定的致病性金黄色葡萄球菌,提取分离菌株的基因组DNA,参考GenBank中发表的ClfA(Clumping Factor A of Staphylococcus aureus)基因序列,设计特异性引物经PCR扩增得到1 560bp的ClfA基因。对新疆3个地区分离的9株金黄色葡萄球菌分别进行ClfA基因的克隆和序列分析,对ClfA蛋白的氨基酸序列分析结果表明,新疆3个地区ClfA蛋白氨基酸的同源性为91.5%～100%,与国外其他分离菌株的同源性为91.9%～100%,差异性较小,该结果表明ClfA基因在该菌进化和流行过程中是保守的。
基金partially funded by the National Natural Science Foundation of China(No.30960411)973 Program(No.2010CB534909)
文摘Objective:To analysis and identify a bacterium strain isolated from laboratory breeding mouse far away from a hospital.Methods:Phenotype of the isolate was investigated by conventional microbiological methods,including Gram-staining,colony morphology,tests for haemolysis, catalase,coagulase,and antimicrobial susceptibility test.The meek and 16S rRNA genes were amplified by the polymerase chain reaction(PCR) and sequenced.The base sequence of the PCR product was compared with known 16S rRNA gene sequences in the CenBank database by phylogenetic analysis and multiple sequence alignment.Results:The isolate in this study was a gram positive,coagulase negative,and catalase positive coccus.The isolate was resistant to oxacillin,methicillin,penicillin,ampicillin,cefazolin,cipr of loxacin erythromycin,et al.PCR results indicated that the isolate was meek gene positive and its 16S rRNA was 1465 bp.Phylogenetic analysis of the resultant 16S rRNA indicated the isolate belonged to genus Saphylococcus,and multiple sequence alignment showed that the isolate was Saphylococcus haemolyticus with only one base difference from the corresponding 16S rRNA deposited in the CenBank.Conclusions: 16S rRNA gene sequencing is a suitable technique for non-specialist researchers.Laboratory animals are possible sources of lethal pathogens,and researchers must adapt protective measures when they manipulate animals.
文摘【目的】探索宁夏干旱荒漠区苦豆子内生放线菌的多样性及区系组成,为苦豆子内生放线菌的合理开发和利用提供理论依据。【方法】从宁夏白芨滩自然保护区不同植被和土壤类型的6个样区采集健康苦豆子样品30份,采用组织匀浆法从苦豆子植株的根部、茎部、叶部和种子中分离内生放线菌,根据培养性状、菌落、孢子等的形态特征和16S r RNA基因序列分析对分离菌株进行鉴定;根据苦豆子内生放线菌的相对频率、物种多样性指数、丰富度指数和相似性系数分析其区系组成特点。【结果】从30份苦豆子样品中,共分离得到内生放线菌111株,这些菌株分属于链霉菌属(Streptomyces)、拟诺卡氏菌属(Nocardiopsis)、普劳斯氏菌属(Prauserella)、Actinophytocola、微杆菌属(Microbacterium)、束丝放线菌属(Actinosynnema)、嗜热油菌属(Thermoleophilum)、糖霉菌属(Glycomyces)和糖丝菌属(Saccharothrix)9个属,其中链霉菌属(Streptomyces)和拟诺卡氏菌属(Nocardiopsis)为优势属。苦豆子内生放线菌的分布具有一定的组织特异性,根和种子中的内生放线菌数量高于茎和叶。植被类型中,沙生植被草原(样区Ⅳ)分离的苦豆子内生放线菌物种多样性指数最高,而荒漠草原(样区Ⅴ)最低。荒漠草原(样区Ⅴ)和林地(样区Ⅵ)内生放线菌群落有密切的相似性。【结论】苦豆子体内含有丰富的内生放线菌资源,其内生放线菌具有很高的宿主特异性,而且其分布受生境影响。
文摘【目的】了解新疆地区牛病毒性腹泻（BVD）的流行状况,【方法】采用RT-PCR方法对从新疆16个不同地区采集的样品进行BVD病毒5＇-UTR扩增,对扩增出的序列进行测序,构建系统遗传进化树和遗传变异分析。【结果】566份样品中,扩增出21份阳性样品,平均阳性率为3.71℅。通过对分离株序列的同源性比对分析,17株为BVDVI型,4株为BVDVⅡ型。经对PCR扩增阳性样品进行细胞传代,有7株为致细胞病变型（CP）,14株为非致细胞病变型（NCP）毒株,所测得的病毒滴度为1.04×103~6.02×108TCID50/0.1 m L,21株分离病毒对BVDV阳性血清的中和效价为1：21~1：25。【结论】BVD在新疆地区均不同程度的发生和流行。研究的成果,为BVD的有效防控提供了技术资料。
基金supported by the Natural Science Foundation Projects in Chongqing(CSTC,2009BB5403)Education Research Project of Chongqing Science and Technology Board (KJ090321)Medical Science and Technology Research Projects of Health Bureau of Chongqing, P.R. China(2009-2-216)
