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为了解山羊致病性大肠杆菌广西分离株的分子多态性,应用随机扩增多态性方法(RAPD)对山羊致病性大肠杆菌进行分型研究.从8条随机引物中筛选出4条能在10株大肠杆菌中具有较好多态性扩增的随机引物,4条随机引物共扩增出18条DNA片段,10个菌株无共有带谱,显示出良好的扩增多态性.菌株Nx31与Nx32曾被认为是同一菌株的两次分离,但是在RAPD分析中,两株细菌的带谱存在明显的差别,表明RAPD比传统的血清学分型具有更高的分辨性. 相似文献
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利用随机扩增多态性DNA指纹技术对猪链球菌进行基因分型 总被引：8，自引：1，他引：7
通过优化反应条件和筛选随机引物，应用100条随机引物对分离的20株猪链球菌做随机扩增多态性DNA（RAPD）分型研究。结果表明，有27条引物呈现良好的多态性和稳定性，可产生稳定，复杂的指纹图谱。采用SPSS10.0软件分析了不同菌株之间的遗传距离，绘制出了菌株间的亲缘关系树状图，结果，20株猪链球菌被分为4个聚类群。 相似文献
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鸭疫里氏杆菌鸭源致病性大肠埃希氏菌和鸭沙门菌的RAPD研究 总被引：5，自引：0，他引：5
以4株不同血清型的鸭疫里氏杆菌、5株不同血清型的鸭源致病性大肠埃希氏菌和5株同一血清型的鸭沙门为研究对象，分别提取基因组DNA，利用随机扩增多态性DNA(RAPD)技术对其基因组DNA进行了分析。结果，从20条随机引物中筛选出了38条能在这3种菌中有较好多态性的随机引物，8个有效引物共扩增出了102个DNA片段，其中14个菌株共有的谱带仅有1条，显示多态性的片段有101个，占99．0％；对RA的4个菌株扩增出的谱带数为51条，共同片段有12条，多态性片段39条，占76．5％；对沙门菌的5个菌株扩增出的条带数为46条，共同片段为13条，多态性片段33条，占71．79／6；对大肠埃希氏菌的5个菌株扩增出的条带数为48条，共同片段4条，多态性片段44条，占91．7％。在8条引物中进一步筛选出1条引物G12，其图谱中535bp的条带为鸭疫里氏杆菌所特有，330bp的条带为沙门菌所特有，1227bp的条带为大肠埃希氏所特有，表明G12可作为分子标记用来鉴别这3种细菌。 相似文献
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鸭沙门菌的随机扩增多态性DNA分析 总被引：5，自引：0，他引：5
以5株同一血清型的鸭沙门茵为研究对象，分别提取基因组DNA后，用20条随机引物对其进行RAPD分析。结果，20条随机引物中不能扩出任何条带的引物有2条，能扩出条带但扩增图谱中没有特异性条带出现的引物有10条，能扩出条带且扩增图谱具有多态性的引物有8条。对筛选到的具有多态性的8条引物进行分析，共扩增出46个DNA片段，片段大小为0．2～3．2kb，其中5个菌株共有的片段有13条，显示多态性的片段有33条。 相似文献
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贵州地区牛源金黄色葡萄球菌分离株16S rRNA分子鉴定及RAPD分型研究 总被引：2，自引：2，他引：0
利用16S rRNA保守序列对引起贵州地区奶牛乳房炎的金黄色葡萄球菌进行分离鉴定,建立随机多态性扩增体系对金黄色葡萄球菌分离株进行基因分型研究。结果表明,69份奶样中共分离出金黄色葡萄球菌37株,DNA随机多态性扩增（random amplified polymorphic DNA,RAPD）结果显示这37株金黄色葡萄球菌扩增产物在1～7条带之间,产物大小为400～4500 bp。菌株可分为7个基因型,其中Ⅰ型7株（18.9%）、Ⅱ型2株（5.4%）、Ⅲ型2株（5.4%）、Ⅳ型1株（2.7%）、Ⅴ型19株（51.4%）、Ⅵ型1株（2.7%）、Ⅶ型5株（13.5%）,Ⅴ型为该地区的流行优势菌群。地理和气候环境对病原菌流行传播的影响是病原菌基因型分布差异的重要原因。 相似文献
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利用RAPD技术对不同地区及不同季节发生危害的柞蚕寄生蝇进行了遗传多态性研究。选取11个随机引物对14份供试材料的基因组DNA进行扩增,共扩增出140条稳定条带,其中128(91.43%)条为多态性条带,表明柞蚕饰腹寄蝇(Blepharipa tibialis)、秋柞蚕寄生蝇(未命名)及栗蚕寄生蝇(未命名)间具有丰富的多态性。各材料间的遗传距离在0.0214~0.5143之间,利用UPGMA法进行聚类分析,14份供试材料可聚为两大类:栗蚕(Dic-tyoplocajaponica)寄生蝇与寄生柞蚕的寄生蝇各聚为一类,其中来自河南省的柞蚕饰腹寄蝇自成一类,来自东北地区的柞蚕饰腹寄蝇与秋柞蚕寄生蝇又各自聚为一类。 相似文献
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采用传统的血清学分型方法和随机扩增多态性DNA分析(RAPD)方法对长春地区分离的猪链球菌进行分型。结果表明，RAPD法比传统血清学分型方法更能反映菌株间的亲缘关系，并从分子水平上揭示了不同菌株的遗传距离；RAPD法具有快速、简便、分型性高等优点。 相似文献
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苜蓿假盘菌ISSR反应体系优化及指纹图谱构建 总被引：4，自引：1，他引：3
以北京苜蓿假盘菌(Pseudopeziza medicaginis (Lib.) Sacc.)菌株为试材,用ISSR-23引物[序列为(AC)8T]研究PCR反应体系的主要成分及退火温度对假盘菌ISSR扩增的影响,以期为苜蓿假盘菌的深入研究奠定基础。结果表明:Primer、Mg2+、Taq酶和dNTP浓度对扩增均有影响,以Primer影响最为明显;优化的反应体系为:20μL反应体系中含0.1 ng模板DNA,2.0mmol/L MgCl2、0.5μmol/L Primer、1U Taq DNA聚合酶、0.2 mmol/LdNTP,2μL 10×PCR Buffer和2.5%去离子甲酰胺;在此基础上,建立6个不同地理来源苜蓿假盘菌菌株的指纹图谱并分析其亲缘关系;对44条ISSR引物进行筛选,22条可产生清晰稳定的多态性条带,其中16条可将6个供试菌株完全分开,建立了苜蓿假盘菌指纹图谱;经聚类分析,在0.55遗传相似水平下,6个供试菌株分为两个遗传谱系,菌株的遗传谱系与其地理来源之间不表现相关性。 相似文献
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鸭茅杂交种的SSR分子标记鉴定及其遗传变异分析 总被引：5，自引：1，他引：4
以140个鸭茅杂交种单株及其亲本(01996、YA02-103)为材料,用SSR分子标记技术研究杂种与其亲本间扩增谱带的多态性,以甄别真假杂种。SSR标记在供试材料中的扩增带型表现为互补型、父本型及其他型。利用3对特异引物A01G20、A03N16、A03K22可快速准确地鉴定出杂交种111份。试验所用的100对SSR引物中有13对引物分别在杂种和双亲之间存在扩增多态性。13对引物在供试材料中共扩增出多态性条带76条,平均每对引物扩增出6条,多态性百分率为84.24%。供试材料具有丰富的遗传变异,利用SSR进行鸭茅杂种鉴定及遗传变异分析是可行的。 相似文献
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The reliability and reproducibility of random amplified polymorphic DNA analysis (RAPD) was compared with restriction fragment length polymorphism (RFLP) by analysing three virus strains isolated from zoo animals in Berlin and three isolates which were cultivated from pets from Northern Germany. The RAPD technique was evaluated as a reliable tool with good reproducibility of the patterns for each virus strain investigated. Problems of interpretation due to inconsistent intensity of bands in different polymerase chain reaction runs may arise for less experienced personnel. The RAPD analysis can be performed within one working day and needs less DNA compared with RFLP so costs will be reduced. The obvious advantage of RFLP is that the pattern can be traced to the recognition site of the restriction enzyme whereas the RAPD primer sequence is not present in the orthopoxvirus genome at all. To the authors knowledge, the RAPD technique has never been applied in DNA viruses before and they conclude that this technique is a useful tool for the discrimination of closely related cowpoxviruses. 相似文献
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为了解绵羊肺炎支原体(Mycoplasma ovi pneumoniae,Mo)在我国部分地区的流行病学特征,采用随机扩增多态性DNA(RAPD)及限制性片段长度多态性(RFLP)方法对26株Mo基因多态性进行了研究,并利用NTsys2.10e软件对获得的多态性图谱进行了聚类分析.结果在相似系数为0.70时,26株Mo可分成6个RAPD群或6个RFLP群;相似系数为0.90时,分成18个RAPD群或18个RFLP群;相似系数为1.00时,分成25个RAPD群或26个RFLP群.结果表明,我国Mo存在高度的基因多态性,这种多态性与地域差异相关,与宿主来源也具有一定的相关性.研究结果为了解我国Mo的分子流行病学特征打下了基础,也为建立有效的Mo诊断方法和疫苗研制提供了参考依据. 相似文献
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随机扩增多态DNA（RAPD）技术在鹅育种上的应用 总被引：10，自引：0，他引：10
本文用4种引物（OPH5、OPH13、OPH16和OPF4）对隆昌鹅、太湖鹅和新太湖鹅进行基因组DNA、RAPD分析。结果表明：三种鹅都有特异性的条带,扩增DNA条带都表现为多态性,条带数为3-12条,大小范围在0.36-0.38kb之间,多态频率为72.14%;RAPD标记可作为一种辅助手段来比较鹅群体的遗传变异性,太湖鹅遗传变异性大于隆昌鹅;OPF-04很可能用作区分隆昌鹅与太湖鹅的标记性引物。 相似文献
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药用植物紫花地丁的分子鉴定及其亲缘关系研究 总被引：2，自引：0，他引：2
运用RAPD技术对传统药用植物紫花地丁（Viola yedoensis）及其7个近缘种进行了分析,筛选出的6个随机引物在8个种39份材料中共扩增出163个位点,其中161个位点具有多态性,占98.7%;在扩增产物中有7条特异性扩增条带可用于紫花地丁的鉴定。根据RAPD结果计算的各种间遗传距离的范围为0.022 7-0.424 4,斑叶堇菜与细距堇菜的亲缘关系最近,它们间的遗传距离最小（0.022 7）,而与紫花地丁遗传距离最近的是早开堇菜,为0.147 1。在所研究的类群中,维西堇菜与裂叶堇菜的遗传距离最大（0.424 4）。根据遗传距离所作的聚类分析结果与紫花地丁等8种植物的形态分化相一致,此外,同一种的不同个体总是先聚在一起,反映了种内不同个体在遗传组成上相对一致性。研究结果表明,RAPD在种内是相对稳定的,能快速、准确地应用于紫花地丁的分子鉴定及其与近缘类群关系的研究。 相似文献
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Rincón G D'Angelo M Gagliardi R Kelly L Llambí S Postiglioni A 《Research in veterinary science》2000,69(2):171-174
Uruguayan Creole cattle inhabit areas that cannot sustain conventional farming. They have adapted to fragile environments and are influenced only by natural selection. In this study, random amplified polymorphic DNA (RAPD) and microsatellite (MS) markers were used to analyse Creole cattle genome polymorphism. A comparative analysis using the RAPD technique was performed in pooled DNA of three cattle breeds (Holstein Friesian, Creole and Hereford) in order to evaluate their amplification patterns. A primary screening of RAPD primers allowed us to select and use those with higher percentage of GC base composition. A total of 215 loci ranging between 300 and 2500 bp were amplified. Bandsharing frequency (BSF) among breeds showed that less related fingerprints were observed between Creole and Hereford cattle (0.77), while the highest similarity frequency corresponded to Holstein Friesian compared to Hereford (0.81). Specific RAPD bands were identified in the three DNA pools and they were tested in every individual of each breed. It may be possible to isolate and sequence these bands to create breed-specific molecular markers. The identification of multiple alleles of the MSCYP 21 in Creole cattle with an heterozygosity of He = 0.846 supported the variability of this genetic resource. The use of molecular markers such as RAPD s and microsatellites is proposed to establish genetic distance among American Creole cattle and possibly related ancestral Iberian breeds. 相似文献
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RAPD-PCR and PFGE as tools in the investigation of an outbreak of beta-haemolytic Streptococcus group A in a Swedish hospital 总被引：1，自引：0，他引：1
González-Rey C Belin AM Jörbeck H Norman M Krovacek K Henriques B Källenius G Svenson SB 《Comparative immunology, microbiology and infectious diseases》2003,26(1):25-35
We evaluated the random amplified polymorphic DNA (RAPD) and pulsed-field gel electrophoresis (PFGE) techniques for studying an outbreak of beta-haemolytic streptococci group A (GAS) occurred at two maternity wards at Danderyd hospital, Stockholm, Sweden. All the isolates were of T-type 8,25. The RAPD technique revealed that all RAPD-PCR profiles were identical. PFGE showed that all the patterns but one were identical. These patterns were compared with 10 different T-type GAS from the strain collection of the Swedish Institute for Infectious Disease Control (SMI) and T-type 8,25 from different years and locations. The SMI strains exhibited patterns different from each other and all different from the isolates from Danderyd hospital. Moreover, RAPD could not differentiate among the T-type 8,25 isolates from different years and locations but PFGE showed differences among the amplicons. Our results indicated that the RAPD and PFGE techniques could be efficient tools in epidemiological studies of GAS. 相似文献
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Ramasoota P Krovacek K Chansiripornchai N Mörner AP Svenson SB 《Acta veterinaria Scandinavica》2000,41(3):249-259
A standardized-reagents commercial kit for random amplified polymorphic DNA (RAPD) analysis was used for typing 58 Escherichia coli strains that were recovered from the milk of sows, having coliform mastitis, within a single swineherd in Sweden. Previously, the 58 E. coli strains were characterized serologically and profiled biochemically. They were also evaluated for their serum resistance and their ability to adhere to fibronectin and bovine fetal fibroblasts. The RAPD analysis was fast, easily performed, and required only a nanogram of DNA. The indistinguishable banding patterns obtained with repeated analyses of 2 isolates from each strain demonstrated that RAPD analysis using standardized beads is a technique that provides reproducible results for typing E. coli strains that cause mastitis in sows. The results of the RAPD analyses demonstrated that E. coli sow mastitis strains are highly variable in serotype, biochemical profiles, virulence factors, and RAPD type, and that all 58 strains can be differentiated by means of the RAPD technique. The strains grouped into 24 RAPD types by combining the results of 2 primers, and into 38 groups by combining the results of serotype and RAPD type. No relationship between serotypes, virulence factors and RAPD types was found. 相似文献