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81.
为研究禽致病性大肠杆菌强毒株E058的毒力相关基因在鸡体内、体外表达情况以及E058和尿道致病性大肠杆菌HEC4在LB和尿液中培养的表达情况,本研究分别提取E058株在SPF鸡体内及E058株和HEC4株在LB和尿液中静置培养的总RNA,与构建的DNA芯片杂交,检测和分析RNA的差异表达情况。芯片的检测结果表明:E058株在鸡体内和LB中培养差异表达基因共有9个,上调基因为5个,分别为neuC、iutA、cvaC、aes-15和iucCD;下调基因为4个,分别为aes-8、gyrB、aec-30和mdh。另外,芯片检测结果也显示E058株和HEC4株在LB和尿液中静置培养,具有相似的基因表达情况。 相似文献
82.
基因芯片方法检测6种动物源性人兽共患病病原 总被引：1，自引：0，他引：1
为了建立可同时检测H5亚型禽流感病毒、狂犬病病毒、猪链球菌2型、炭疽芽孢杆菌、沙门氏菌、大肠杆菌O157的基因芯片检测方法,本实验根据GenBank中上述6种病原的基因序列,设计并合成了特异性的引物和探针.采用点样法制备杂交芯片,将上述病原扩增产物混合后与芯片杂交.杂交结果显示,针对本试验中6种人兽共患传染病所设计的寡核苷酸探针可特异性识别靶基因,与其他常见病原体之间没有交叉反应.检测的灵敏度在1.38×10-5pg/μL～151 pg/μL之间.将建立的基因芯片检测方法对临床样品进行检测,结果与荧光PCR方法一致. 相似文献
83.
Using different typing methods (MLST, spa‐, SCCmec‐ and agr‐typing), PFGE and DNA microarray‐based chip analysis, we characterized 20 MRSA strains isolated from livestock and veterinarians. PFGE analysis after macrorestriction with EagI provided seven different band patterns, which could be grouped into four clusters. One cluster consisted of all MRSA ST398 strains isolated from pigs, calves, mastitis milk and two veterinarians. One strain of ST398 from a veterinarian and the two strains of ST1 and ST8 formed the three other clusters. Antimicrobial susceptibility testing showed that 15 of 20 strains were resistant to ampicillin, cefoxitin, clindamycin, erythromycin, oxacillin, penicillin and tetracycline. All strains were susceptible to rifampin and vancomycin, 19 were susceptible to ciprofloxacin and 18 were susceptible to sulphamethoxazole/trimethoprim. Genes encoding different enterotoxins, leukotoxins and haemolysins were found in certain strains. 相似文献
84.
应用基因芯片技术检测禽重要疫病的研究——I．4种禽病检测基因芯片靶基因的克隆及鉴定 总被引：3，自引：3，他引：0
对NDV、IBV、AIV和IBDV等病原进行了分子生物学特征分析，以RT-PCR分别扩增制备NDV、IBV、IBDV和AIV的靶基因，分别命名为ND1、ND2、ND3、ND4、ND5、IB1、IB2、IB3、IB4、AI1、A12和IBD1。并分别对制备的靶基因进行了克隆和鉴定分析，经BamHⅠ酶切、PCR和核酸序列测定鉴定分析表明，除T／WZ重组质粒外，T／ND1（F48E9），T／ND2（LaSota），T／ND3（LaSota），T／ND4（F48E9），T／ND5（LaSota），T／ND5（F48E9），T／IB1（SM），T／IB1（M41），T／IB2（SM），T／IB3（M41），T／WZ，T／AI1，T／AI2，T／IBD1等14个重组质粒，均是其病原的特异性基因。该批靶基因的成功克隆和鉴定为禽疫病检测基因芯片的构建和制备提供了确实可靠的靶基因材料，是禽类疫病基因检测芯片构建和制备的关键技术之一。 相似文献
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LIU Li-na LIU Zheng-fei RUAN Li WANG Zhao-xiong LI Ren-feng ZHANG Song-lin CHEN Huan-chun HE Qi-gai 《中国农业科学(英文版)》2007,6(2):234-243
DNA microchip used in this study was formed from miniature arrays of pseudorabies virus （PrV） gene-specific probes immobilized on a glass surface. Hybridization using DNA microchip （microarrays） was used for differentiation between virulent and attenuated PrV. The presence of four gene segments （gB, gD, gE~, and gE ） encoding conservative glycoprotein B （gB）, D （gD）, and E （gE） of PrV was monitored using multiplex PCR. The amplicons were labeled with Cy5 or Cy3 dyes followed by hybridization to the gene-specific capture probes on the microchip. The presence of gD and gB, gE~ gene fragments was shown in virulent （gE~ genotype） and attenuated PrV （gE genotype）, whereas gE- gene （deleted domain in gE gene） was demonstrated only in virulent, not in attenuated, virus. No cross-hybridization was observed when fluorescence labeled-PCR products of PrV were hybridized using capture probes of related viruses, such as porcine respiratory and reproductive syndrome virus （PRRSV）, porcine parvovirus （PPV）, Japanese encephalitis virus （JEV）, and porcine circovirus type 2 （PCV-2）. The assay was 10 times sensitive than gD gene-specific PCR. Overall, the results of this study suggested that the microarray might be very useful for detection and differentiation of virulent PrV from attenuated one. 相似文献
88.
Giovanna Jona Lasinio Alessio Pollice Livia Pappalettere Giovanni Vannacci Sabrina Sarrocco 《Plant pathology》2021,70(5):1146-1157
The Biolog phenotype microarrays (PM) system offers a simple and cheap tool to rapidly provide a high throughput of information about the phenotypes of fungal isolates in a short time. In order to improve the use of the PM system in fungal ecology studies, the present work proposes a new statistical protocol based on two approaches, that is, a functional principal components analysis to describe similarity patterns of growth curves, and a Bayesian generalized additive model (GAM) to allow inferences on specific growth features, in order to analyse nutrient fungal utilization in a model system including four causal agents of Fusarium head blight, the natural competitor Fusarium oxysporum, and the beneficial isolate Trichoderma gamsii T6085. Analysis of data collected by the Biolog PM in our biological system showed a different nutritional competitive potential of the four pathogens, as well as an intermediate behaviour of the natural competitor and of our biocontrol agent. This protocol, applicable to different fungal phenotypical studies at both isolate and community level, allows a full exploitation of data obtained by the PM system and provides important information about the nutritional pattern of a single isolate compared to those of other fungi, a key factor to be exploited in biocontrol strategies. 相似文献
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