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核酸检测技术是目前动物疫病诊断和食品安全监测的重要手段。多重核酸检测技术可同时检测多种病原,具有通量高、损耗低的优势,在疫病诊断和食品安全监测等领域的应用越来越广泛。目前已研发出多种多重核酸检测技术,包括聚合酶链式反应(polymerase chain reaction,PCR)多病原核酸检测技术、环介导等温扩增(loop-mediated isothermal amplification,LAMP)多病原核酸检测技术、生物芯片多病原核酸检测技术、高通量测序多病原核酸检测技术等。由于检测原理不同,不同方法之间的检测性能存在较大差异。本文通过概述基于PCR/RT-PCR、LAMP/RT-LAMP、生物芯片、高通量测序建立的多重核酸检测技术,全面分析其在动物疫病诊断和食品安全监测领域中的特点和具体应用策略,以期推进病原检测技术的创新和发展,为动物疫病诊断及食品安全监测提供技术支持。 相似文献
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针对鸭圆环病毒病毒诊断,目前已经建立了PCR、Real Time PCR、电镜观察、核酸探针技术、间接ELISA方法等方法. 相似文献
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PCR即聚合酶链反应是利用DNA聚合酶在体外大量扩增两段已知序列之间的某一特定DNA片段(又称模板)的分子生物学技术.这种技术可以利用几乎所有的临床样品探查病原体基因的存在和变异,从而对生物体的状态和疫病作出诊断.因而,如何从被检临床样品中高效、简便地提取核酸作为模板,就成为普及、推广PCR技术检测动物疫病亟待解决的问题.现依据笔者操作和学习体会,将能有效检测动物疫病的样品种类及处理方法介绍如下,以图共同发展PCR检测动物疫病技术. 相似文献
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目前,在基层兽医实验室中实验人员常用实时荧光PCR方法对动物疾病作定性诊断,而病毒核酸提取方法的选择影响荧光PCR实验结果的准确性和效率。本实验选择屠宰检疫过程中较常见的细菌性病原猪丹毒杆菌和病毒性病原猪圆环2型病毒作为研究对象,对热提取法和离心柱吸附法两种核酸提取效果的准确性、稳定性和差异性作对比,在实验人员进行实时荧光PCR实验时选择合适DNA病原提取方法有一定参考意义。 相似文献
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鹅细小病毒和番鸭细小病毒双重PCR检测方法的建立 总被引：1，自引：0，他引：1
根据GenBank上登录的鹅细小病毒(GPV)和番鸭细小病毒(MDPV)基因序列,分别设计合成针对GPV非结构蛋白(NS)和MDPV NS2-VP1基因片段的2对引物GPV U/L和MDPV U/L,将GPV和MDPV提取核酸混合后作为模板,优化PCR反应条件,建立了能同时检测这2种病毒的双重PCR。特异性试验结果显示,引物GPV U/L仅特异性扩增出GPV-GZ1和GPV-GZ2株730bp核酸片段,引物MDPVU/L仅特异性扩增出MDPV的624bp核酸片段,双重PCR扩增出长度分别为730bp和624bp的2条特异性片段,而扩增鸭瘟病毒(DPV)和鹅副黏病毒(GPMV)的核酸扩增结果均为阴性。敏感性试验结果显示,双重PCR能同时检测到14.4pg的GPV核酸和28.8pg的MDPV核酸。结果表明,建立的双重PCR可用于GPV和MDPV的鉴别诊断和联合检测。 相似文献
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核酸探针和PCR技术在食品检验中的应用 总被引：14，自引：0，他引：14
核酸探针技术和多聚酶链反应( Polymerase chain reaction,PCR)技术是近年来发展起来的两种高新生物技术。本文着重介绍了核酸探针技术和 PCR技术的原理及其在食品检验中的应用。同时提出了这两种技术在食品检验中尚存在的问题以及解决的办法 ,并对其在食品检验中的应用前景作了展望。 相似文献
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数字PCR技术在动物疫病检测中的应用 《畜牧与饲料科学》2021,42(6):124-128
为了支持动物疫病流行地区的控制及消除计划,以及在动物疫病非流行地区进行有效筛查,必须使用更加准确灵敏的诊断手段。数字PCR可实现绝对定量及痕量核酸的检测,与传统检测方法相比具有不依赖参考物或标准品、对抑制剂具有较高耐受性、灵敏度和精准度更高的优点。数字PCR作为一种定量分析的核酸检测新方法,在动物疫病检测中得到了应用,围绕数字PCR技术的原理、应用、存在的问题及发展趋势等进行综述及分析,为数字PCR在动物疫病检测中的进一步应用提供参考。 相似文献
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选择中国大陆最早分离的H9N2亚型禽流感病毒(avian influenza virus,AIV)A/Chicken/Guangdong/SS/94(H9N2)(缩写为SS株)和1998年大流行时期分离的H9N2亚型AIVA/Chicken/Shanghai/F/98(H9N2)(缩写为F株)为研究对象,对其在SPF鸡体内的复制能力和传播途径特性比较后发现,F株在4周龄SPF鸡气管中的复制能力高于SS株,F株可以经气溶胶传播途径传播,SS株不能经气溶胶传播途径传播;利用反转录-聚合酶链反应(RT-PCR)方法获取F株和SS株的HA和NA基因的cDNA,序列分析得知,F株和SS株的HA和NA基因的同源性分别是96.6%和98.1%;HA基因的裂解位点氨基酸序列都是PARSSR↓GL,但有5个氨基酸的差异,即166位N(F)→D(SS)、198位A(F)→V(SS)、217位V(F)→I(SS)、335位G(F)→R(SS)、504位L(F)→S(SS);2株病毒的NA基因在63~65位都存在氨基酸缺失,但在NA基因红细胞吸附位点的氨基酸序列不同,分别是IKKDSRSG(F)和IKEDLRSG(SS)。F株和SS株的传播特性差异是否与其表面基因序列有关,有待进一步研究。 相似文献
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禽类的起源、演化及我国主要家禽品种类型与分布 总被引：1，自引：1，他引：0
家禽是重要经济价值动物.本文从禽类种群进化学说出发,简介了禽类的起源、演化、动物学分类和家禽的驯化(养)与品种的形成,并对我国主要家禽(鸡、鸭、鹅)地方品种和培育品种(配套系)的分布与类型作了描述,以期为研究我国家禽起源系统,保护与利用我国家禽品种,促进家禽生产可持续发展提供参考. 相似文献
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近年以来,由于市场因素的刺激,生猪的存养量大幅上升,再加上由于流通环节较多,流通非常频繁,流通距离越来越远。这对繁荣经济,增加养殖效益起了重要的推动作用,但也同时给疾病的感染和传播创造了有利条件,给猪病的防治带来了困难。有的猪场感染了传染病后,由于治疗不及时不得法,而造成了惨重的经济损失。2008年7月中旬,我街道一养猪户因盲目从外地购进中猪,发生猪病疫情,引起猪只连续死亡,造成一定的经济损失。根据流行病学、临床症状、剖检变化和实验室诊断,诊断该病为猪链球菌病和猪伪狂犬病混合感染,现报告如下。 相似文献
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１前言１．１鸡白冠病鸡白冠病是由卡氏住白细胞原虫寄生于鸡的红细胞和单核细胞而引起的鸡的贫血性疾病。吸血昆虫蚋和库蠓叮咬鸡引起传播，是主要的传播媒介，一般在夏末和秋季多发，由于夏季降雨量较大，部分沟渠积水，库蠓和蚋多孳生，因此在多雨水涝的年份发病率明显增高。１９９８年中国从南到北发生洪涝灾害，吸血昆虫的孳生格外严重，出现了一个白冠病多发年，而后两年发病稍轻，并有地区性，今年８月中旬以来白冠病的发病呈抬头趋势，有一定的死亡率，对蛋鸡产蛋率也会引起一定程度的降低，应引起养鸡户的重视。１．２鸡痘鸡痘也是… 相似文献
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Pinard CL Weiss ML Brightman AH Fenwick BW Davidson HJ 《American journal of veterinary research》2003,64(1):104-108
OBJECTIVE: To evaluate lactoferrin and lysozyme content in various ocular glands of bison and cattle and in tears of bison. SAMPLE POPULATION: Tissues of ocular glands obtained from 15 bison and 15 cattle and tears collected from 38 bison. PROCEDURE: Immunohistochemical analysis was used to detect lysozyme and lactoferrin in formalin-fixed, paraffin-embedded sections of the ocular glands. Protein gel electrophoresis was used to analyze ocular glands and pooled bison tears by use of a tris-glycine gel and SDS-PAGE. Western blotting was used to detect lactoferrin and lysozyme. RESULTS: Immunohistochemical staining for lactoferrin was evident in the lacrimal gland and gland of the third eyelid in cattle and bison and the deep gland of the third eyelid (Harder's gland) in cattle. Equivocal staining for lactoferrin was seen for the Harder's gland in bison. An 80-kd band (lactoferrin) was detected via electrophoresis and western blots in the lacrimal gland and gland of the third eyelid in cattle and bison, Harder's glands of cattle, and bison tears. An inconsistent band was seen in Harder's glands of bison. Lysozyme was not detected in the lacrimal gland of cattle or bison with the use of immunohistochemical analysis or western blots. Western blots of bison tears did not reveal lysozyme. CONCLUSIONS AND CLINICAL RELEVANCE: Distribution of lactoferrin and a lack of lysozyme are similar in the lacrimal gland of cattle and bison. Differences in other tear components may be responsible for variability in the susceptibility to infectious corneal diseases that exists between bison and cattle. 相似文献
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Harvey CE 《Journal of veterinary dentistry》2002,19(4):186-195
Crown width, height and buccal surface areas were measured on heads or skulls of four dogs and four cats, and were compared with similar measurements on models of human dentition. Buccal surface area variability was greater in dogs and cats than in humans, and teeth of cats were smaller. Horizontal (gingival and occlusal halves) and vertical (mesial, middle, and distal thirds) buccal surface area variability was also greater in canine and feline teeth compared with human teeth. This increased variability suggests the need for testing of reliability and repeatability of scoring when using plaque and calculus indices based on horizontal or vertical segmentation. Buccal surface area variability between teeth also prompts questioning the validity of equal weighting of smaller, irregularly-shaped teeth when calculating a mean mouth score. Whether equal or more reliable results would be obtained from scores of whole teeth in comparison with segmentation indices used currently has yet to be determined. 相似文献
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Over a period of about 12 years, 30 abnormal Schistosoma mattheei cercariae were found among a total of approximately 2.8 million examined. Initially seven were recovered from about 1.02 million (0.0007%), which were examined individually while being counted with the aid of a stereoscopic microscope. Subsequently, on the strength of relatively high percentages of abnormal individuals recovered when counting cercariae that failed to penetrate into oxen, it appeared that the morphologically abnormal cercariae were unable to swim and would mostly sediment out of a suspension while most of the normal cercariae would remain swimming. This surmise is supported by recovery of 23 morphologically abnormal cercariae (0.001%) from about 1.8 million, by examining the sediment after the cercarial suspension had been left standing undisturbed in glass measuring cylinders. The abnormalities ranged from aberrant tails only (e.g. an underdeveloped tail, or different degrees of schism) or aberrant heads only, to abnormalities of both the heads and tails. A suggested schematic classification of abnormal cercariae is presented. A young, adult hamster was exposed to eight S. mattheei cercariae with complete schism of the shaft of the tail, by pipetting the cercariae onto the shaved abdominal skin of the anaesthetised animal. Two underdeveloped females were subsequently encountered in squash preparations of the liver when the hamster was killed for worm recovery 10 weeks after infection, thus showing that some of the abnormal cercariae were viable. A method is also described for killing and fixing cercariae while retaining some of the shining brilliance of live cercariae, without them becoming shrivelled, granular and semi-opaque, as occurs when cercariae die spontaneously or are killed with heat. This is apparently the first report of abnormal cercariae of S. mattheei. In addition, a method of concentrating abnormal cercariae after emergence from a snail, a schematic classification of abnormal cercariae and a method for killing and fixing cercariae while retaining much of the shiny brilliance of live cercariae are also reported for the first time as far as is known. 相似文献