共查询到19条相似文献，搜索用时 671 毫秒
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环介导等温扩增技术（LAMP）是由Notomi等于2000年开发的新颖的恒温核酸扩增方法,在等温条件下可高效、快速、高特异、高灵敏地扩增靶序列。1技术特点环介导等温扩增技术特点是针对靶序列的6个特异性区域设计两对引物,利用具有链置换活性的DNA聚合酶在恒温条件下30～60分钟,不需要PCR仪即可实现核酸的大量扩增,并且伴有肉眼可见的副产物白色焦磷酸酶沉淀产生。LAMP的扩增效率是目前最高的一种, 相似文献
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NASBA （Nucleic acid sequence-based amplifi-cation） 技术是由加拿大Cangen公司于1991年提出的一种核酸序列依赖扩增技术,该技术是由一对特异的引物介导的、三种酶催化的以单链RNA为模板的恒温扩增技术,具有操作简单、特异性强、灵敏度高、不易被污染等优点, 相似文献
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NASBA技术研究进展 总被引：1，自引：0，他引：1
NASBA （Nucleic acid sequence-based amplifi-cation） 技术是由加拿大Cangen公司于1991年提出的一种核酸序列依赖扩增技术,该技术是由一对特异的引物介导的、三种酶催化的以单链RNA为模板的恒温扩增技术,具有操作简单、特异性强、灵敏度高、不易被污染等优点, 相似文献
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核酸依赖性扩增技术（NASBA）是继PCR技术后发展起来的一门新型的体外核酸扩增技术。它的特点是：恒温、高效、特异、不需要特殊的仪器设备。本文就现阶段核酸依赖性扩增技术的特点及其在动物疫病检测中的应用情况和前景作一综述。 相似文献
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Anais Prouteau Florian Chocteau Clotilde de Brito Edouard Cadieu Aline Primot Nadine Botherel Frdrique Degorce Laurence Cornevin Marie A. Lagadic Florian Cabillic Pauline de Fornel‐Thibaud Patrick Devauchelle Thomas Derrien Jerome Abadie Catherine Andr Benoît Hdan 《Veterinary and comparative oncology》2020,18(2):214-223
Canine oral melanoma is the first malignancy of the oral cavity in dogs and is characterized by a local invasiveness and a high metastatic propensity. A better knowledge of genetic alterations is expected to improve management of this tumour. Copy number alterations are known characteristics of mucosal melanomas both in dogs and humans. The goal of this study was to explore the prognostic value of somatic focal amplifications on chromosomes (Canis Familiaris [CFA]) 10 and 30 in canine oral melanoma. The cohort included 73 dogs with oral melanoma confirmed by histology, removed surgically without adjuvant therapy and with a minimal follow‐up of 6 months. Epidemiological, clinical and histological data were collected and quantitative‐PCR were performed on formalin‐fixed paraffin‐embedded (FFPE) samples to identify specific focal amplifications. The 73 dogs included in the study had a median survival time of 220 days. Focal amplifications on CFA 10 and 30 were recurrent (49.3% and 50.7% of cases, respectively) and CFA 30 amplification was significantly associated with the amelanotic phenotype (P = .046) and high mitotic index (MI; P = .0039). CFA 30 amplification was also linked to poor prognosis (P = .0005). Other negative prognostic factors included gingiva location (P = .003), lymphadenomegaly (P = .026), tumour ulceration at diagnosis (P = .003), MI superior to 6 mitoses over 10 fields (P = .001) and amelanotic tumour (P = .029). In multivariate analyses using Cox proportional hazards regression, CFA 30 amplification (Hazard ratio [HR] = 2.08; P = .011), tumour location (HR = 2.20; P = .005) and histological pigmentation (HR = 1.87; P = .036) were significantly associated with shorter survival time. Focal amplification of CFA 30 is linked to an aggressive subset and constitutes a new prognostic factor. 相似文献
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鉴别牛早期胚胎性别PCR方法引物的设计与筛选 总被引：6，自引：2，他引：6
根据牛Y-染色体特异重复序列、睾丸特异蛋白基因以及性别决定基因序列设计合成5对公牛Y-染色体特异引物，依据牛骨胳肌α肌动蛋白前体基因和微卫星DNA序列设计合成4对牛DNA特异引物(内标引物)。单重PcR扩增牛基因组DNA，筛选出4对牛Y-染色体特异引物和1对牛DNA特异内标引物。将不同的Y-染色体特异引物与内标引物组合，多重PCR扩增牛基因组DNA、已知性别的牛成纤维细胞和克隆胚胎，筛选出2个可用于牛早期胚胎性别鉴别的PCR引物组合：B34／A12和B78／A12。 相似文献
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本试验根据GenBank中登录的禽传染性贫血病毒(CAV)基因序列,设计合成2对引物,外引物的扩增片段大小为485 bp,内引物的扩增片段大小为297 bp,建立了适合CAV快速检测的套式PCR方法(nested PCR),并采用该方法对CAV阳性毒株及临床病料进行了检测。结果显示,该方法能扩增到297 bp的条带,禽流感病毒(H9亚型)、新城疫病毒、传染性法氏囊病病毒、禽网状内皮增生病病毒、减蛋综合征病毒、禽呼肠孤病毒、马立克氏病病毒的扩增结果均为阴性。该方法第1步扩增的敏感性是100 pg,第2步PCR扩增的敏感性是1 fg,敏感性提高了105倍。本研究建立的CAV套式PCR方法具有敏感性高、重复性好、特异性强等优点,可用于CAV的临床诊断和分子流行病学调查等。 相似文献
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本试验根据GenBank中登录的禽白血病病毒(ALV)基因组序列,设计合成了2对引物,外部引物的扩增片段大小为478 bp,内部引物的扩增片段大小为314 bp,建立了适合ALV快速检测的套式PCR方法(nested-PCR)。采用该方法对ALV毒株进行了检测,试验结果表明,能扩增到314 bp的条带,禽流感病毒(H9亚型)、新城疫病毒、传染性法氏囊病病毒、减蛋综合征病毒、禽网状内皮增生病病毒、禽呼肠孤病毒、马立克氏病病毒的扩增结果均为阴性。该方法第1次扩增的敏感性是100 pg,第2次扩增的敏感性是1 fg,第2次比第1次扩增的敏感性高105倍。所建立的套式PCR方法具有敏感性高、重复性好、特异性强等优点,可用于禽白血病病毒(ALV)的临床诊断和分子流行病学调查等。 相似文献
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According to the sequence of hexon gene of fowl adenovirus groupⅠ(FAVⅠ) strain published in GenBank,two pairs of primers were designed and synthesized.The outer primers amplified a fragment of 475 bp in length, and the inner primers amplification fragment was 237 bp in length. A nested PCR assay for rapid detection of FAVⅠ was established.A specific 237 bp fragment was amplified from DNA templates of FAVⅠstrain,but no bands were amplified with templates extracted respectively from avian influenza virus (AIV) subtype H9,Newcastle disease virus (NDV), infectious bursal disease virus (IBDV),duck plague virus (DPV), reticuloendotheliosis (REV), avian reovirus (ARV), Marek's disease virus (MDV). Sensitivity of the 1st and 2nd amplifications by the nested PCR assay were 100 pg and 1 fg,respectively.The sensitivite of the 2nd amplifications increased by 105 times.The results showed that the nested PCR was specific,sensitive,rapid,accurate,and could be used as a routine assay for the detection of FAVⅠ.This method had good reproducibility, specificity and sensitivity, and might detect low content FAVⅠ accurately and rapidly. This method could be used as a method for the diagnosis and detection of clinical cases,and molecular epidemiological investigation of FAVⅠ. 相似文献