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农药残留超标已成为影响农产品质量安全的重要问题,迫切需要探寻开发灵敏、准确、可靠、便捷且适用性强的农药残留快速检测方法。免疫层析法是将抗原抗体特异性免疫反应和色谱层析分离技术相结合的一种快速检测方法,其中,基于胶体金标记的免疫层析技术以其便捷、成本低、可视化等优点而受到普遍欢迎。近年来随着量子点、时间分辨荧光微球、上转换发光纳米粒子等新型纳米标记材料的出现,免疫层析技术得到了广泛发展。文章从标记类型（非共价作用标记及共价作用标记）及标记材料（胶体金、纳米碳、量子点、上转换发光纳米粒子、磁性纳米颗粒、时间分辨荧光微球及荧光乳胶颗粒）等方面,综述了不同纳米材料标记的免疫层析技术及其在农药残留检测领域的研究及应用进展,可为深入开展农药残留免疫层析技术研究提供参考。 相似文献
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一种新型病毒检测技术 总被引：1，自引：0，他引：1
一种新型病毒检测技术（河北农业技术师范学院河北昌黎０６６６００）刘品贤有关植物病毒的诊断和鉴定方法是很多的，常用的有免疫扩做法、凝聚法、酶联免疫鉴定法（ＥＬＩＳＡ）、荧光抗体法、直接荧光诊断法（ＤＦＤ）、免疫吸附电镜法等。应用上述方法对病毒进行检测，... 相似文献
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为研究细菌磁颗粒分离提取不同样本材料中的植物病毒RNA的效果,以及结合实时荧光 RT-PCR技术检测LSV的灵敏性,选取感染病毒的西葫芦叶片（CGMMV和SqMV）、大豆种子（BPMV）与百合叶片（LSV和ArMV）3种样本材料,利用细菌磁颗粒分别提取这5种病毒RNA,与Trizol方法提取效果进行比较,同时与Trizol real-time RT-PCR检测LSV的灵敏度进行了比较。结果表明：BMPs方法能够从3种植物样本中提取病毒RNA,其检测LSV的灵敏性与Trizol方法相当。 相似文献
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花期干旱对大豆叶绿素荧光参数的影响 总被引：4，自引：0，他引：4
利用叶绿素荧光动力学测定技术,研究大豆开花期干旱对叶片叶绿素荧光参数的影响,结果表明:大豆开花期受旱后,可变荧光与最大荧光比(Fv/Fm)、可变荧光与初始荧光比(Fv/F0)、非光化学淬灭系数(NPQ)均降低,而ETR升高。说明光系统Ⅱ(PSⅡ)受到了伤害,使得PSⅡ原初光能转换效率(Fv/Fm)、PSⅡ潜在活性(Fv/F0)、起光保护作用的热耗散降低,光合电子传递速率升高。花期干旱胁迫后,各参数存在基因型差异,新大豆1号在花期干旱条件下,光合机构受破坏较轻,其吸收的光能能较多地用于光化学转化能力,抗旱能力强。因此,花期干旱胁迫下,叶绿素荧光参数(Fv/F0、Fv/Fm)变化与大豆品种抗旱性有关,利用大豆品种叶绿素荧光对干旱胁迫的反应差异鉴定品种抗旱性是可行的。 相似文献
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苜蓿花叶病毒(alfalfa mosaic virus, AMV)是一种世界性分布、宿主范围广、具有严重危害性的植物病毒,能引起大豆的严重病害。本研究利用原核表达的AMV CP蛋白制备的抗血清,建立了高效、准确的AMV间接ELISA检测方法,并应用于病害调查和抗性鉴定,结果表明制备的3份抗血清对重组蛋白和AMV感染的大豆植物粗提液的效价均达到256 000倍,血清特异性分析结果显示3份抗血清仅识别感染AMV的大豆叶片,不识别感染大豆花叶病毒(soybean mosaic virus, SMV)的大豆叶片。通过建立的AMV间接ELISA与常规RT-PCR同时对采集的50份疑似感染AMV的大豆样品进行检测,有46份样品检测结果一致,符合率达92%。利用建立的AMV ELISA方法和课题组已建立的SMV ELISA方法对吉林省大豆主产区的大豆样品进行病毒检测的结果表明,病毒检出率为38.30%,SMV的检出率达30.85%,AMV的检出率达17.06%,复合侵染率为9.61%。对接种AMV的40个大豆品种进行抗性鉴定,结果显示40份大豆全部感染AMV,但是病毒载量存在差异,部分品种表现出AM... 相似文献
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李剑锋 《中国进出境动植检》1995,(2):30-31
番茄环斑病毒的检测方法一般有下列几种: 一、生物学检测 生物学方法是植物病毒检测的最常用方法之一,将植物病毒接种到指示植物(Indicator plant)上,通过指示植物的特殊症状,来判定是哪一种病毒。将TomRSV的PSP分离物(桃茎纹孔病分离物)摩擦接种于以下9种诊断寄主上,2—3周后出现以下症状: 苋色黎(Chenopodium amaranticolor):小的局 相似文献
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繁殖与推广植物检疫签准的无毒种子、无性繁殖材料是防止病毒传播、控制水果、蔬菜和观赏作物的许多严重病毒病害的根本方法。几十年来,植物病毒学家已经用免疫化学技术来探测和分类植物病毒。自从1977年引用酶联免疫吸附测试(ELISA)于物植病毒的检测以来,血清学测定方法在植物检定中已广为应用。高质量抗血清的需求大增。在许多检定程序中血清学测定显得日趋重要。为提高血清学方法的灵敏度,血清的质量必须改进,避免非专化性反应出现。在植物病毒提纯过程中,某些植物材料总是附随于病毒,也作为抗原而刺激产生其专化性抗体反应。血清学测试越有效,每次测试需要的抗体量越少,寄主抗体的低含量也就显得越重要。许多在凝胶双扩散测试中能用的血清,除非先用健康植物汁液交互吸收过,否则不适合于做ELISA反应;某些情况下,即使是吸附收过的抗血清在ELISA反应中仍不能用。三十多种植物病毒的单克隆抗体(MCAs)已能制备了,在植物病毒的基础研究和应用研究中MCAs都有许多大用途。现在许多MCAs已应用于病毒检测和病毒间相互关系的研究。MCAs在研究根本性问题上也是有用的:如病毒怎样装配?为何病毒感染某种植物而不感染其它种植物?病毒怎样克服那结合到农作物中的抗性?在植物体中病毒是怎样运动的?在阐述病毒感染过程中的复杂生化反应时MCAs也是有用的。本文将集中谈谈用MCAs检测病毒和未来几十年病毒检测中MCAs将在哪些方面进行改进。 相似文献
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应用MNP-RT-PCR方法检测黄瓜绿斑驳花叶病毒 总被引：4，自引：0，他引：4
A novel RT-PCR method integrated with Magnetic Nano Particles (MNP), MNP-RT-PCR, was set up for detection of Cucumber green mottle mosaic virus (CGMMV). After the virus particles in crude sap were concentrated by MNP, viral RNAs were released and were detected by RT-PCR. CGMMV could be detected in as less as 10 ng watermelon leaf materials. Compared with normal RT-PCR, the method decreased the inhibitors of plant material and steps for extracting RNA, and also increased the sensitivity of RT-PCR detection in less time. The method is simple and suitable for quick detection of plant virus in a large number of samples. 相似文献
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采用磁性纳米微球增强信号的表面等离子体共振免疫传感系统检测水中莠去津残留 总被引：1，自引：0，他引：1
将磁性纳米微球(MNP,表面修饰羧基的磁性四氧化三铁微球)与表面等离子体共振(SPR)免疫传感技术结合,以莠去津单克隆抗体(AT-m Ab)与磁性纳米微球的偶联物(AT-m AbM NP)作为传感识别元件,初步建立了一种用于饮用水中除草剂莠去津残留检测的SPR信号增强免疫传感方法。通过对检测条件的优化,该方法对自来水中莠去津的检出限为0.89 ng/m L(S/N=3),检测范围为8.62~7.18×10~3ng/m L,检测时间小于20 min;在10~1 000 ng/m L添加水平内,莠去津的平均回收率为94%~102%,相对标准偏差(RSD)为5.1%~7.3%。磁性纳米微球的加入有效增强了SPR传感器的响应信号强度,提高了检测方法的灵敏度。本研究建立了一种快速、灵敏、准确的水中莠去津残留的检测方法,可为相应的现场检测技术和设备的研发提供技术基础。 相似文献
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In order to develop a rapid, sensitive and specific qPCR assay for detection and quantification of Tomato yellow leaf curl virus (TYLCV), a pair of primers and TaqMan probe were designed according to the conserved sequence of known TYLCV isolates. Combining with MNP technique, a novel MNP-qPCR detection method was established and verified based on specificity, sensitivity and reproducibility tests. The results indicated that the Ct value of plotted standard curve showed good linear relationship(R2 =0.9994)with the log of copy number of template. The established method showed a high specificity for TYLCV detection without crossing reaction with Tomato severe leaf curl virus and Tomato yellow leaf curl Sadinia virus, and was 10-fold more sensitive than routine PCR. Both coefficients of variation were less than 2%, indicating a good reproducibility. We have provided a novel method for detection of TYLCV in plant samples rapidly and quantitatively. 相似文献
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以"富士"苹果树为试材,对果树体生物量和各器官及其皮层、木质部的磷含量和累积动态进行了研究,以期探讨果树磷素吸收、转运和分配规律.结果表明,3月26日至9月21日,树体生物量呈直线增加,果实采收后,生长缓慢,7月30日后,根系快速生长,植株、地上部及根系中磷累积迅速增加;枝、干和根系皮层内磷含量与累积7月30日最低,休眠期最高,皮层磷含量枝＞干＞根系;而木质部磷含量与累积4月30日达最低,木质部磷含量根系＞枝＞干;一年内,果树吸磷总量为28.72 kg/hm2,果实和叶片共带走磷素7.94 kg/hm2,7月30日至9月21日,吸收磷素18.32 kg/hm2,占吸收总量的63.8%,9月21日至1月15日,吸收磷素10.40 kg/hm2,占36.2%;苹果树(苹果产量48 t/hm2)年推荐施纯磷47.86 kg/hm2,果实收获后秋季基施磷17.33 kg/hm2,果实膨大期前追施磷30.53 kg/hm2. 相似文献
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为探究长期定位施肥对黄土高原渭北旱塬麦田土壤线虫群落的影响,以长武国家黄土高原农业生态试验站的长期定位试验（1984—2018年）为平台,调查裸地（L）以及不施肥（CK）、施氮磷肥（NP）、单施有机肥（M）、氮磷肥配施有机肥（MNP）的小麦田土壤线虫群落数量、组成结构和生态功能指数,并分析其与土壤理化性质的关系。结果表明：（1）小麦田土壤线虫数量显著高于裸地,而CK、M和MNP处理使小麦田中土壤线虫数量比NP处理提高了31.63%~56.20%;（2）施肥减缓了长期种植小麦导致的食细菌线虫（特别是头叶属）相对丰度的下降,单施有机肥的处理（M）植食性线虫相对丰度显著降低,同时,相比于CK,施用有机肥的M和MNP处理杂食/捕食线虫相对丰度分别增加了18.4%和8.24%,表明有机肥对杂食/捕食线虫的促进作用;（3）长期施肥处理土壤线虫Shannon多样性指数（H）和指示土壤健康状况的瓦斯乐卡指数（WI）分别为1.80~2.19和0.36~0.68,相较于裸地（H=2.36;WI=1.57）有所下降,但M和MNP处理的线虫成熟度指数（MI）和结构指数（SI）均高于其他处理（L、CK和NP处理）,表明施用有机肥有助于土壤食物网维持复杂的结构和成熟稳定的状态;（4）不同施肥措施下土壤全氮、有机碳、微生物生物量碳和氮以及可溶性碳的变化是影响土壤线虫数量和群落特征的重要环境因素。 相似文献
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Neil Boonham Rachel Glover Jenny Tomlinson Rick Mumford 《European journal of plant pathology / European Foundation for Plant Pathology》2008,121(3):355-363
The detection and identification of plant pathogens currently relies upon a very diverse range of techniques and skills, from
traditional culturing and taxonomic skills to modern molecular-based methods. The wide range of methods employed reflects
the great diversity of plant pathogens and the hosts they infect. The well-documented decline in taxonomic expertise, along
with the need to develop ever more rapid and sensitive diagnostic methods has provided an impetus to develop technologies
that are both generic and able to complement traditional skills and techniques. Real-time polymerase chain reaction (PCR)
is emerging as one such generic platform technology and one that is well suited to high-throughput detection of a limited
number of known target pathogens. Real-time PCR is now exploited as a front line diagnostic screening tool in human health,
animal health, homeland security, biosecurity as well as plant health. Progress with developing generic techniques for plant
pathogen identification, particularly of unknown samples, has been less rapid. Diagnostic microarrays and direct nucleic acid
sequencing (de novo sequencing) both have potential as generic methods for the identification of unknown plant pathogens but
are unlikely to be suitable as high-throughput detection techniques. This paper will review the application of generic technologies
in the routine laboratory as well as highlighting some new techniques and the trend towards multi-disciplinary studies. 相似文献
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ABSTRACT Phytophthora ramorum is a recently described pathogen causing bleeding cankers, dieback, and leaf blight on trees and shrubs in parts of Europe and North America, where the disease is commonly known as sudden oak death. This article describes the development of a single-round real-time polymerase chain reaction (PCR) assay based on TaqMan chemistry, designed within the internal transcribed spacer 1 region of the nuclear ribosomal (nr)RNA gene for detection of P. ramorum in plant material. Unlike previously described methods for the molecular detection of P. ramorum, this assay involves no post amplification steps or multiple rounds of PCR. The assay was found to have a limit of detection of 10 pg of P. ramorum DNA, and could detect P. ramorum in plant material containing 1% infected material by weight within 36 cycles of PCR. The assay also was used to test DNA from 28 other Phytophthora spp. to establish its specificity for P. ramorum. A quick and simple method was used to extract DNA directly from host plant material, and detection of P. ramorum was carried out in multiplex with an assay for a gene from the host plant in order to demonstrate whether amplifiable DNA had been extracted. Amplifiable DNA was extracted from 84.4% of samples, as demonstrated by amplification of host plant DNA. The real-time protocol was used to test 320 plant samples (from 19 different plant species) from which DNA extraction had been successful, and was shown to give results comparable with a traditional isolation technique for diagnosis of P. ramorum in plant material from common U.K. hosts. 相似文献
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The detection of Ralstonia solanacearum (biovar 2A) in stems of symptomless plants before harvest of the potato crop, instead of tubers, would not only save highly valued planting material but would be less time-consuming and would also enhance farmers' market decisions. Although pathogen detection in stems has been proven efficient for ring rot, this has never been investigated for bacterial wilt (BW). Therefore the possibility of detecting BW latent infection in stem pieces about three weeks before harvest was assessed in 57 fields of the Andean highlands of Peru. Two sensitive, specific and user-friendly serological methods were used to detect the pathogen in latently infected tubers and stems: double-antibody sandwich (DAS)-ELISA and indirect ELISA on nitrocellulose membrane (NCM) after enrichment of the plant extracts in a semi-specific broth. Optimum sample sizes of stems and tubers were evaluated for 37 potato crops showing between 0 and 0·1% BW incidence using a binomial distribution model to calculate the detection probabilities. Although results of detection using the two serological techniques had 100% concordance, detection probabilities were higher using DAS-ELISA, whatever the plant part tested. BW detection probabilities were higher for tubers than for stems; a 99% detection probability was obtained by analysing 400 stems sections or 250 tubers using DAS-ELISA. Detection of BW infection in symptomless plants 20 days before harvest using post-enrichment DAS-ELISA is a reliable and user-friendly technique that can easily be used by national plant protection services and seed programmes in developing countries. 相似文献