猪肺炎支原体PCR检测方法的建立及初步临床应用     

张红云  梁晶晶  李回  孟先明  潘艳  冯励  罗廷荣 《广西农业科学》2010,41(3):256-258

根据GenBank中猪肺炎支原体（Mhp）J株P36蛋白基因（登录号X67286）的核苷酸序列设计1对引物,建立了快速检测Mhp的PCR方法。该方法能扩增出948bp的Mhp特异性条带,其敏感性达到可检测出0．735ng的MhpDNA,但对鸡毒支原体、猪伪狂犬病毒、猪圆环病毒、猪流感病毒、猪呼吸与繁殖综合征病毒均未检测出相应目的条带;将克隆获得的目的片段与GenBank已发表的MhpJ株的P36基因进行比较,同源性达到99．9％。采用所建立的PCR方法对42份疑似猪支原体肺炎（MPS）病料进行临床诊断,发现有13份呈阳性（31．0％）。可见,该PCR方法适合于MPS的临床诊断,可为其防控提供可靠依据。  相似文献   

Isolation of Mimosine Degrading Bacteria from Rumen Juice and Mass Production by Gottingen Bioreactor Technology     

A. Aung  U. ter Meulen  F. Gessler  H. Bohnel 《农业科学与技术》2011,(5):764-772

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鸡场饮水中肺炎克雷伯氏菌的分离鉴定及耐药性研究     

符德佳  康桦华  曾宪军  樊志红  陈杰  黄炜乾  蒋顺进  彭新宇 《广东农业科学》2024,51(2):124-138

【目的】研究养殖场鸡饮水中肺炎克雷伯氏菌（Klebsiella pneumoniae）的耐药表型和耐药基因的携带情况,并分析其相关性,为有效预防该病提供参考。【方法】从养殖场鸡饮水系统中分离、鉴定肺炎克雷伯氏菌,采用药敏纸片扩散法检测分离菌株对25种常用抗菌药的耐药性,通过PCR检测菌株携带的常见耐药基因。对其中4株肺炎克雷伯氏菌进行全基因组测序,利用CARD抗性基因数据库预测耐药基因的携带情况。【结果】从养殖场鸡饮水系统中共分离到9株肺炎克雷伯氏菌,药敏试验表明这些菌株呈现出多重耐药,仅对多粘菌素敏感。PCR检测发现,9株肺炎克雷伯氏菌共携带13种常见的耐药基因,其中,β-内酰胺酶类耐药基因CMY-1、BlaSHV和BlaCTX-M,碳青霉烯类耐药基因KPC,喹诺酮类耐药基因QnrB、QnrS和QnrA,氨基糖苷类耐药基因Aph(3’)-Ia,磺胺类耐药基因Sul2等12种耐药基因与其耐药表型表现一致,而Mcr-1基因与其表型相反。全基因组扫描分析结果显示,4株肺炎克雷伯氏菌共携带了59种耐药基因,涉及31类抗生素药物,其中四环素类抗生素的耐药基因最多,其次是大环内酯类抗生素和氟喹诺...  相似文献   

产气克雷伯氏杆菌 Klebsiella aerogenes S2对植物病原真菌的抑菌活性及番茄促生效果     

杨淼泠#  王威#  申乃坤  李时勇  黄文善  施李鸣  姜明国  张克诚  葛蓓孛 《植物保护》2022,48(5):91-98

本研究以广西红树林的根际土壤为材料, 利用选择性培养基分离筛选多功能菌株, 并利用生理生化和分子生物学相结合的方法进行鉴定?采用平板对峙法测定筛选获得的菌株对几种植物病原真菌的拮抗效果, 同时通过盆栽试验和发芽试验研究其对番茄幼苗和种子的促生效果, 从而挖掘兼具促生防病功能的植物根际促生菌(plant growth-promoting rhizobacteria, PGPR)?经筛选获得一株具有溶磷及分泌生长素(indole-3-acetic acid, IAA)能力的细菌菌株S2, 经鉴定该菌株属于克雷伯氏菌属Klebsiella, 其对禾谷镰孢Fusarium graminearum, 玉蜀黍平脐蠕孢Bipolaris maydis等3种植物病原真菌具有一定的拮抗效果, 菌株发酵液浓度为1×104?2×104 cfu/mL时对番茄种子和幼苗的促生效果最好?综上, 菌株S2 是一株新型且兼具促生防病效果的多功能PGPR菌株, 该结果为绿色现代农业开发土壤微生物菌肥和植物生长调节剂提供优质菌源参考?  相似文献   

The role of chlamydia pneumoniae in the course of transformation of C57 BL/6J mouse peritoneal macrophages into foam cells     

JIN Jun-fei  YANG Yong-zong 《园艺学报》2000,16(8):730-733

AIM: In order to understand the role of chlamydia pneumoniae in the course of macrophages transformation into foam cells experiments with chlamydia pneumoniae standard strain AR-39 wese made. METHOD: C57 BL/6J Mouse peritoneal macrophages C57 BL/6J wese incubated 24 h, and they were divided into 6 groups to be incubated continually: A₁~DMEM; A₂~DMEM+10 IFUs/L AR-39; B₁~DMEM+10mg/L LDL; B₂~DMEM+10mg/L LDL+10 IFUs/L AR-39; C₁~DMEM+10mg/L OxLDL;C₂~DMEM+10mg/L OxLDL+10 IFUs/L AR-39. 72 h later, morphological changes of the cells were observed and of intracellular cholesterol of content was detected. RESULTS: 1、Morphological studies: there were no lipid particles in A₁, A₂ and B₁ groups, but the lipid particles could be found in B₂、C₁ and C₂ group, Among six groups, the most lipid particles were seen in C₂ groups. 2、Biochemical detection:The ratio of cholesterol ester to total cholesterol was much higher than 50% in B₂、C₁ and C₂ groups, but was less than 50% in A₁、A₂ and B₁ groups. CONCLUSION: Chlamydia pneumoniae may have played a role in promoting C57 BL/6J mouse peritoneal macrophages into foam cells.  相似文献   

水貂源肺炎克雷伯杆菌分离、鉴定及疫苗潜能分析     

张国军  王殿永  杨艳梅  陈建国  畅丽芳  吴金  高玉龙  楚洪源  柳增善  张媛媛 《西北农业学报》2022,(10):1243-1249

水貂在夏季养殖时常出现出血性肺炎症状,病原大部分为铜绿假单胞菌。然而近10 a来采自河北、山东、辽宁等地水貂部分出血性肺炎病例中没有分离到铜绿假单胞菌,为探清病原,针对以上病料进行其他病原的分离鉴定。通过病料的细菌分离、常规鉴定,16S rRNA基因测序,动物回归试验、毒力试验、免疫原性试验及攻毒保护试验等,确定6株分离菌株有致病性,并被鉴定为肺炎克雷伯杆菌,分别命名为FY1、FY2、FY3、FY4、FY5、FY6,其中FY3为毒性最强的肺炎克雷伯氏菌(ZC1108株)。以该毒株对10月龄健康水貂进行肌肉注射,结果显示其致死率为100%(5/5);以该菌株制备的灭活疫苗免疫水貂后攻毒保护率达100%(5/5),试验水貂全部健活。表明肺炎克雷伯氏菌(ZC1108株)灭活疫苗免疫原性良好。  相似文献   

Establishment and Preliminary Application of Triple PCR to Identify Three Kinds of Conditional Pathogenic Bacteria     

FENG Jie  XIE Jian-yun  HU Jian-hua  GAO Cheng 《中国畜牧兽医》2015,42(6):1389-1395

The aim of this study was to establish a simultaneous triple PCR detection method for Staphylococcus aureus (S.aureus),Pseudomonas aeruginosa (P.aeruginosa) and Klebsiella pneumoniae (K.pneumoniae).Three pairs of specific primers had been designed according to nuc gene of S.aureus,toxR gene of P.aeruginosa and PhoE gene of K.pneumoniae.The triple PCR reaction conditions were optimized on the basis of single PCR methods.At the same time,specificity,sensitivity and repeatability tests of the triple PCR method were studied,and the results of bacteria isolation and culture and the triple PCR method were compared.The results showed that the amplification product sizes were 484,278 and 368 bp,respectively.The optimal annealing temperature was 56 to 59 ℃,the concentrations of primers were all 0.2 μmol/L,dNTP concentration was 200 μmol/L,Mg²⁺ concentration was 2.5 mmol/L.The specificity test showed that there was no cross reaction between these three bacteria templates and other eight kinds of common bacteria templates,such as Bordetella bronchiseptica.The minimum of simultaneous detection of three bacteria genomic DNA was 10^-5 ng/μL.The results of three repeatability tests of triple PCR were the same which indicated the repeatability was good.30 samples from mice were detected by bacteria isolation and culture and the triple PCR method,the detection rate of triple PCR was slightly higher than the bacteria isolation and culture.The positive samples detected by bacteria isolation and culture were also positive detected by triple PCR.The results showed that a specific,sensitive and efficient triple PCR system had been established and could provide the technical support for bacteria detection and epidemiological investigation of laboratory animals.  相似文献   

肺炎支原体感染大鼠实验模型的建立   总被引：1，自引：0，他引：1  

李继昌  董龙  杨阳 《畜牧与兽医》2004,36(8):16-18

本试验旨在建立肺炎支原体感染的Wistar大鼠模型 ,为研究肺炎支原体的发病机制及药物治疗理论提供基础。采用滴鼻法对实验大鼠进行肺炎支原体感染 ,利用PCR法进行咽拭子检测 ,并以透射电镜和光学显微镜进行肺部病理组织学检查。结果发现 ,实验大鼠在感染肺炎支原体 1 0d时 ,咽拭子PCR检测结果为阳性 ;透射电镜观察到肺脏细胞膜破裂 ,线粒体变性 ,嵴断裂 ;光镜下可见到支气管及肺血管周围有明显的淋巴细胞浸润 ,形成斑片状间质性支气管肺炎。结果表明了Wistar大鼠对肺炎支原体较易感 ,并产生以间质性肺炎为主要特征的肺部和呼吸道感染 ,也说明本次造模试验成功  相似文献   

基于非理性设计的肺炎克雷伯氏杆菌甘油脱水酶的分子定向进化     

陈云来  韦旭钦  杜丽琴  韦宇拓  黄日波 《广西农业生物科学》2014,(1):42-50

1,3-丙二醇是一种具有广泛应用前景和巨大市场潜力的平台化合物。在生物体转化甘油合成1,3-丙二醇的代谢途径中,甘油脱水酶是代谢途径的限速酶,然而自然界中的甘油脱水酶却存在着诸多的不足。本研究采用易错PCR技术对来源于肺炎克雷伯氏杆菌的甘油脱水酶进行非理性设计的分子定向进化,通过高通量筛选方法筛选获得了一个酶学性质得到了改善的突变酶γ-F97G。当以甘油作为酶催化的底物时,突变酶的酶比活力为（583.8±49.4）U/mg,相比较野生型的酶比活力提高了约94%;当以1,2-丙二醇作为酶催化底物时,突变酶的酶比活力为（147.9±5.5）U/mg,相比较野生型的酶比活力提高了约72%。经分析酶蛋白的三维结构后发现酶蛋白发生突变的位点属于远距离突变位点,实验结果表明远距离突变有时同样是一条优化和改善酶的酶学性质的好途径。  相似文献   

混合感染背景下的猪气喘病控制     

邵国青 《中国兽药杂志》2009,43(10):49-52

混合感染背景下的猪气喘病较难控制,介绍了猪气喘病的发病特点,常用药物的敏感度评价、疫苗效果评价和猪场猪气喘病的检查检测方法,结合作者实际经验强调了饲养管理要点,指出了种猪场净化猪气喘病的方法步骤,为混合感染下控制猪气喘病提供参考。  相似文献