共查询到19条相似文献，搜索用时 265 毫秒
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环介导等温扩增（loop-mediated isothermal amplifi cation,LAMP）技术是一门新兴的分子生物学检测技术,因其具有特异性强、敏感性高、简单、快捷及不需要昂贵的仪器设备等特点,受到了生物医学研究者的高度关注。目前,该方法已经被广泛应用于各种病原微生物检测。本文综述了近年来LAMP在动物疫病检测中的具体应用实例和目前的研究进展,并对LAMP方法在动物疫病诊断中的应用前景做了展望。 相似文献
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《中国畜牧兽医文摘》2016,(1)
环介导等温扩增技术(Loop-mediated Isothermal Amplification,简称LAMP)是一种体外等温扩增核酸片段的新技术,利用4条特异性引物,在Bst DNA聚合酶的催化下能够有效地扩增出靶基因。与常规PCR法相比该法具有特异性强、灵敏度高、快速准确以及操作简便等优点,现已广泛应用于动物疾病检测中。此文综述了LAMP技术原理及其在鸡传染性疾病病原检测中的应用,旨在为该技术的深入研究和应用提供参考。 相似文献
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变性高效液相色谱高通量检测动物源性饲料中沙门氏菌和志贺氏菌 总被引：5，自引：0，他引：5
应用复合PCR结合变性高效液相色谱(DHPLC)技术,通过通用增菌培养,建立动物源性饲料中沙门氏菌和志贺氏菌的快速高通量检测方法.根据沙门氏菌和志贺氏菌特异基因序列分别设计特异性引物,复合PCR扩增的产物经DHPLC技术进行快速检测.以49株参考菌株做特异性试验,并开展了精密度检测试验.试验结果表明,该方法具有很好的特异性和精密度,经通用增菌和复合PCR-DHPLC技术可同时检测饲料中的沙门氏菌和志贺氏菌.该方法可以快速、准确、高通量检测,是动物源性饲料中致病菌高通量检测的新技术. 相似文献
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分子检测技术是进行病原检测、鉴定和疾病诊断的重要手段。重组酶聚合酶扩增技术（recombinase polymerase amplification，RPA）是一种新型核酸扩增技术，主要在能结合单链核酸（寡核苷酸引物）的重组酶、单链DNA结合蛋白（SSB）和链置换DNA聚合酶3种酶的作用下等温扩增DNA或RNA，具有操作便捷、特异性强、灵敏度高和反应迅速等优点，可实现对疾病的现场诊断。作者简述了RPA技术的反应体系组成、反应原理、引物和探针的设计以及扩增产物的检测方法等，介绍了RPA技术在动物重要病毒性、细菌性及寄生虫病原检测中的应用，分析了该技术的局限性，并对其发展前景进行了展望。 相似文献
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环介导等温扩增技术在动物病原检测中的应用 总被引：1，自引：0，他引：1
环介导等温扩增技术（LAMP）是一种依赖于自动循环的链置换技术,在等温条件下,不到1h就能扩增出10^9-10^10靶序列拷贝。该技术快速准确、操作简单、特异性强、灵敏度高,已经被用于动物病原微生物的快速检测。本文就LAMP法的反应原理、技术特点及其在动物病原快速检测中的应用作一综述。 相似文献
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本研究利用3种方法对新城疫病毒(Newcastle disease virus,NDV)进行检测,并对这3种检测方法的灵敏度和特异性作出比较。根据新城疫病毒的融合蛋白基因(F基因)设计合成6条特异性引物,利用水浴LAMP法、PCR及LAMP实时浊度仪进行检测。通过对恒温水浴中的反应温度、反应时间及反应液中Mg2＋浓度进行优化获得最佳反应条件,结果用凝胶电泳分析;用优化的温度进行LAMP实时浊度仪检测,同时都与PCR方法进行比较。结果显示,获得了最佳反应条件,水浴LAMP方法最低检测限是1.58 pg,比PCR高100倍,而LAMP实时浊度仪检测灵敏度比水浴LAMP方法高10倍,最低检测限是0.158 pg。3种方法对其他非新城疫病毒均无检出。结果表明,新城疫病毒水浴LAMP检测方法速度快、不需要高精密的仪器,而且具有灵敏度高、特异性强、操作简单等特点,有望在核酸扩增领域取代PCR技术,LAMP浊度仪法检测灵敏度更高,但是所需仪器和试剂比较昂贵。 相似文献
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LUO Ya-kun LIANG Lin WANG Jing LIU Cun LIU Qi LIU Chang LIN Wen-cheng CUI Shang-jin 《中国畜牧兽医》2017,44(2):344-349
This study was aimed to establish a loop-mediated isothermal amplification (LAMP) assay for detection of porcine epidemic diarrhea virus (PEDV), and provide a simple, sensitive, accurate and reliable tool for diagnosis of PEDV.The conservative PEDV N gene (GenBank accession number: KT799997) of PEDV was selected as a target to design six specific primers.The reaction system and temperature of LAMP were optimized, and the LAMP method for specific amplification of PEDV was established. Results showed that the PEDV LAMP detection method was established successfully, and it could detect PEDV specifically at 60℃ for 60 min,and the detection limit was 91 copies/μL, which was one hundred-fold higher than conventional RT-PCR method.75 clinical samples were detected by LAMP and PCR, respectively, the coincidence of LAMP and PCR was about 97.3%. All the data suggested that the LAMP assay had strong specificity, high sensitivity, simple operation, low equipment requirement, and was suitable for rapid detection of PEDV clinical samples. 相似文献
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试验旨在建立一种快速检测猪流行性腹泻病毒（PEDV）的环介导等温扩增方法（LAMP）,为诊断PEDV提供简便、敏感、准确可靠的工具。参考GenBank中PEDV基因序列（登录号：KT799997）,针对PEDVN基因设计了6条引物,对所建立的LAMP反应体系、反应温度进行优化,建立可特异性扩增PEDV的LAMP方法。结果显示,本试验成功建立了PEDV LAMP检测方法,在60℃恒温下反应60 min,能特异性地检测PEDV,检测限量为91拷贝/μL,比常规PCR方法的敏感性高100倍。对比75份临床样本的LAMP和常规RT-PCR法检测结果,显示两种方法符合率为97.3%。综上所述,本试验建立的LAMP方法具有特异性强、敏感性高,操作简单,设备要求低的特点,适用于PEDV临床样本的快速检测。 相似文献
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A loop-mediated isothermal amplification (LAMP) assay was developed for detection of Salmonlla in fecal samples of experimental monkeys. According to the specific sequences (fimY) of Salmonlla in GenBank, one set of primers was selected and the reaction condition was optimized. The results showed that the detection limit of LAMP method was 1.35×101 and 1.35×103 CFU/mL in Salmonella pure culture and clinical samples,respectively,which was the same as routine PCR. The amplification could complete in one hour, and the result could be distinguished by naked eyes. There was non-specific amplification of other pathogens. These results suggested that this LAMP assay was a simple and specific method for rapid detection of Salmonlla in fecal samples. 相似文献
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Xie Z Tang Y Fan Q Liu J Pang Y Deng X Xie Z Peng Y Xie L Khan MI 《Avian diseases》2011,55(4):575-579
A loop-mediated isothermal amplification (LAMP) assay was optimized for the rapid detection of Group I avian adenoviruses. A set of six primers was designed from the DNA sequences of hexon genes from Group I avian adenovirus. The assay was performed in a water bath for 60 min at 63 C, and the amplification result was visualized by adding a fluorescence dye reagent or by inspecting the white sediment. The results showed that the LAMP assay could detect all 12 serotypes of Group I avian adenovirus and nine Guangxi Group I avian adenovirus isolates. This avian adenovirus Group I-specific LAMP assay could detect 238 copies of avian adenovirus. No cross-reactions were detected using the LAMP assay with avian adenoviruses type II and III or with other avian viruses. The ability of LAMP to detect Group I avian adenovirus isolates was further evaluated with 184 cloacal swab samples from poultry. In total, 72 out of 184 cloacal swab samples from poultry were identified as positive by LAMP, whereas 45 out of 184 were identified as positive by conventional PCR test. The Group I avian adenovirus specific LAMP results were further confirmed by real-time PCR. This specific LAMP method holds promise as a rapid and specific diagnostic assay for detection of samples from birds suspected of adenovirus infection. 相似文献
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猪圆环病毒2型LAMP检测方法的建立与评价 总被引：1，自引：0，他引：1
根据猪圆环病毒2型(Porcine circovirus type 2,PCV2)Rep和Cap基因保守区设计2对引物,1对外引物和1对内引物;利用设计的4条引物,在Bst大片段聚合酶的作用下,对PCV2 DNA进行恒温扩增;扩增条件为63℃恒温反应1h;建立了PCV2环媒恒温扩增技术(LAMP)检测方法。对检测方法特异性评价表明,检测方法只能检测PCV2 DNA,对猪圆环病毒1型(Porcine circovirus type 1,PCV1)、猪细小病毒(Porcine parvovirus,PPV)及猪伪狂犬病病毒(Porcine pseudorabies virus,PRV)检测无交叉反应。灵敏度评价表明,该检测方法可以检测到样品中10个拷贝的PCV2 DNA含量。 相似文献
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The assay was aimed to establish a rapid, sensitive and specific method of loop-mediated isothermal amplification (LAMP) for the detection of Salmonella. According to the highly conserved STN gene of Salmonella reported in GenBank, we designed a set of primers, and then followed by a series of LAMP optimization of reaction conditions, specificity test, sensitivity test, stability test and enzyme test. The sensitivity of LAMP and conventional PCR were compared.The results showed that the optimal reaction temperature of LAMP was 65 ℃, when only amplification of Salmonella, the minimum detectable concentration was 101 CFU/mL, which was more than conventional PCR detection by one order of magnitude. The results showed that LAMP method had the qualities of specificity, high sensitivity, short time-consuming and convenience for the detection of Salmonella, which was expected to develop into an effective method 相似文献