DNA芯片技术及应用

生物芯片是指通过机器人自动打印或光引导化学合成技术在硅片、玻璃、凝胶或尼龙膜上制造的生物分子微阵列。根据分子间特异性相互作用的原理 ,将生命科学领域中不连续的分析过程集成于芯片表面 ,构建微流体生物化学分析系统 ,以实现对细胞、蛋白质、基因及其它生物组分的准确、快速、大信息量的检测。按照芯片上固定的生物分子的不同 ,可以将生物芯片划分为基因芯片或ＤＮＡ芯片、蛋白质芯片及芯片实验室 (Ｌａｂ -ｏｎ -ｃｈｉｐ)。而从其功能不同的角度 ,生物芯片又可以分为测序芯片、表达芯片和ＣＧＨ芯片。基因芯片是生物芯片研究中 ,…     

生物芯片技术在中药新药开发中的应用前景

综述了生物芯片技术在中药开发中的应用,主要包括药物靶标的寻找、生物指纹图谱的应用、蛋白质芯片的研究以及中药复方的研究.     

基因芯片技术的发展与应用

1 技术原理 生物芯片技术是根据分子间特异性的相互作用原理,将生命科学领域中不连续的分析过程集成于硅芯片或玻璃芯片表面的微型生物化学分析系统,以实现对细胞、蛋白质、基因及其他生物组分的准确、快速、大信息量的检测.     

液相蛋白芯片技术及在人畜疾病诊断中的应用

液相芯片技术是20世纪90年代中期发展起来的被喻为后基因组时代的芯片技术,属于生物芯片的一种。目前,由于传统的固相生物芯片操作繁琐、信息质量稳定性和可重复性差的缺点,极大地限制了芯片技术在疾病诊断领域的应用。液相芯片技术是一种新型生物分子高通量检测技术,这种技术将流式检测技术与芯片技术有机地结合在一起,使生物芯片反应体系由液相-固相反应改变为接近生物系统内部环境的完全液相反应体系。     

基因芯片技术检测细菌耐药性

1　基本概念生物芯片 (Biochip)是指将大量的生物分子探针以大规模阵列形式排布在很小的载玻片、尼龙网等载体上 ,通过与标记的样品进行杂交 ,检测杂交信号的强弱进而判断样品中靶分子的数量。依据生物分子探针的不同 ,生物芯片可分为很多种类 ,包括基因芯片、蛋白质芯片、细胞芯片、组织芯片。目前研究最成功的且形成一定商业市场的是 DNA芯片 (DNAchip或DNA Microarry) ,又称为基因芯片 (Gene chip) ,即将无数预先设计好的寡核苷酸、c DNA、基因组 (Ge-nomic) DNA在芯片上做成点阵 ,与样品中同源核酸分子杂交。包括两种模式 :一是…     

信息之窗

我国研制成功监测肉类药残智能系统用于检测肉类兽药残留的生物芯片检测系统日前在生物芯片北京国家工程研究中心研制成功,这是世界上第一个能够监测肉类中兽药残留的生物芯片系统。据介绍,该生物芯片能够分析大量的生物分子,快速准确地完成肉类中兽药残留的检测工作。已研制成功的这套兽药残留蛋白质免疫芯片检测系统,能够在几分钟内检测出肉类中的兽药残留。目前,该芯片的第一代样品已经研制成功,可以对包括瘦肉精、磺胺二甲嘧啶、链霉素在内的几种重点兽药进行检测。信息之窗农业部:将建立瘦肉精等违禁药品省际互查制度今年农业部将建立…     

科技动态

肉类药残监测智能系统研制成功用于检测肉类兽药残留的生物芯片检测系统日前在生物芯片北京国家工程研究中心研制成功,这是世界上第一个能够监测肉类中兽药残留的生物芯片系统。据介绍,这套兽药残留蛋白质免疫芯片检测系统,能够分析大量的生物分子,在几分钟内检测出肉类中的兽药残留。由于大部分兽药是小分子化学物质,检测时用快速提取仪提取食品中样品,并与抗体混合放在芯片上,抗原、抗体发生竞争免疫反应,再用激光共焦扫描仪检测,最后通过特定的软件自动判断残留是否超标。目前该芯片的第一代样品已经研制成功,可以对包括瘦肉精、磺胺二甲…     

可视化芯片技术研究进展

生物芯片具有准确、快速、高通量等优点而被广泛应用于生物学的各个领域。但芯片的设备的体积较大、价格昂贵、实验室环境要求高,普通实验室难以开展相关工作,在一定程度上制约了生物芯片的应用范围。可视化芯片技术是在常规芯片基础上发展起来的一种技术类型,可分为可视化蛋白芯片和可视化基因芯片。可视化芯片应用新的标记方法取代传统的荧光标记,并借助相应的显色方法,使得试验结果能直接被肉眼所观察到。可视化生物芯片具有快速、可视、设备要求低等特点,近些年来被广泛应用于病原检测、基因突变检测及差异表达、酶功能研究等方面,具有潜在的应用前景。     

蛋白质芯片技术在传染病检测中的应用研究进展

蛋白质芯片以其特异、灵敏及高通量等特点已在生命科学的多个领域得到应用,如蛋白质组学研究、新药的开发、酶与底物的相互作用和疾病检测等。论文详细介绍了蛋白质芯片技术的原理、芯片介质的种类及蛋白质的固定技术,论述了蛋白质芯片在传染病检测中的研究概况,分析了蛋白质芯片检测技术中存在的问题及应用前景。     

应用组织芯片免疫组化法检测ALV-J

禽白血病J亚群（ALV-J）是英国的Payne和他的同事们在20世纪90年代初从肉鸡中分离出来的新亚群，主要引起肉鸡的骨髓瘤白血病。自1999年，我国一些肉用型种鸡场陆续发生了禽白血病J亚群。并且近几年来，ALV-J已从最初只引起肉种鸡发病开始向蛋鸡及中国地方种鸡蔓延。组织芯片技术是一种新型特殊的生物芯片技术，它能明显提高工作效率，减少实验误差。本研究将组织芯片技术和免疫组化染色结合起来，用特异性抗ALV-J囊膜蛋白gp85的单克隆抗体来检测发病鸡只的各组织器官的组织切片。在肝脏、脾脏、肾脏、卵巢、腺胃、骨髓、髓细胞瘤组织均检出病毒阳性抗原。结果表明组织芯片技术和免疫组化染色相结合为临床诊断ALV-J提供了一个高通量、敏感的检测方法。     

生物芯片及其在乳品检测中的应用研究进展

生物芯片作为一种新兴起的微量和高通量分析与检测技术,逐渐开始应用于生物研究、食品、医疗和农业领域等.本文主要介绍生物芯片的基本原理、分类和制作过程.深入探讨生物芯片在食品安全领域中的应用,重点介绍了生物芯片在乳品行业中的应用,并对该技术存在的问题及发展趋势进行了简要的阐述.     

生物芯片技术的特点及其在兽医科学中的应用研究进展

生物芯片技术是21世纪的前沿生物技术,该技术是继基因克隆、基因测序和PCR技术后的又一次革命性的技术突破,现已成为生物技术领域的研究热点之一,它的最突出特点是高通量、高集成、微型化、平行化、多样化和自动化。正是这些优点,使得生物芯片技术日益广泛应用于生命科学研究的众多领域。文章主要对生物芯片的特点和现状、技术环节及其在兽医科学研究中的应用作一综述。     

Detection of leptospires in tissue using an immunoperoxidase staining procedure

An immunoperoxidase technique for the localisation of leptospires in sections of formalin fixed paraffin embedded kidney tissue is described. The procedure utilises a two-layered antibody sandwich with rabbit anti-leptospiral immunoglobin. Using antiserum to specific leptospiral serovars the presence and distribution of specific serovar in the tissue could be determined. The technique was also used to detect leptospires of given serovars in smears made from infected tissues and fluids. There was good correlation between culture results and results of the immunoperoxidase staining method on kidneys infected with leptospires. The diagnostic possibilities of the technique on formalin fixed tissue specimens are discussed.     

Schistosoma mansoni antibodies in tissue specimens demonstrated by a diffusion-in-gel ELISA technique.

A modification of the diffusion-in-gel enzyme-linked immunosorbent assay (DIG-ELISA) for analysis of antibodies in tissue fragments has been elaborated. In this technique, tissue specimens are placed on top of a thin gel layer covering a plastic surface coated with an antigen preparation. The gel thus serves as a medium through which diffusion of antibodies contained in tissue samples may occur. Specimens from skin, lung, liver, kidney, heart and skeletal muscle as well as blood serum and blood sampled onto filter paper were successfully used for analysis of the antibody response to two different antigens (soluble adult worm antigen and soluble egg antigen) during experimental infection of mice with the helminth parasite Schistosoma mansoni. It was concluded that skin and lung tissue reflected the serum antibody response more closely than the other tissues tested, although zone sizes were slightly smaller than those registered for corresponding serum samples. The DIG-ELISA reaction zone size was, as expected, shown to be dependent on the weight (size) of the tissue specimen. The variability of the modified technique was comparable to that found for analysis of serum by the conventional DIG-ELISA technique. It is concluded that the described technique may be used for serological analysis when blood samples cannot be easily obtained, e.g. at examination of animal carcasses or single organs.     

Detection of type II avian adenoviral antigen in tissue sections using immunohistochemical staining.

An immunohistochemical staining technique for the detection of marble spleen disease (MSD) viral antigens and other type II avian adenoviral antigens was developed using a mixture of monoclonal antibodies produced against hemorrhagic enteritis (HE) virus and a commercial streptavidin-biotin peroxidase indicator system. This technique was applied to both frozen and formalin-fixed paraffin-embedded tissue sections. The immunohistochemical staining technique was used on tissues from pheasants with experimental MSD, on tissues from a pheasant with natural MSD, and on tissues from turkeys with natural HE. Staining results were compared with routine hematoxylin-and-eosin (H&E) staining. Additional viral inclusions, not detected with H&E, were found in the liver, lung, bone marrow, and kidney sections using the immunohistochemical technique. The immunohistochemical technique was highly specific and sensitive for the detection of type II adenoviral antigen, and it appears to be useful for studying the pathogenesis of these diseases and for retrospective evaluation of routinely processed diagnostic tissue samples.     

家蚕组织中钙的亚细胞定位研究

应用锑酸盐沉淀技术研究家蚕中肠和马氏管的亚细胞钙离子分布，其螯合试验和电镜观察证明，用本试验方法产生的锑酸盐沉淀是锑酸钙，沉淀分布有规则，超微结构保存良好，是研究蚕体组织亚细胞钙离子分布较为有效的方法。作者应用这一技术观察了家蚕５龄幼虫中肠及马氏管组织的钙离子亚细胞分布，并发现氟中毒后马氏管组织的钙离子亚细胞分布有显著的变化，认为这一技术可为家蚕氟毒理研究提供一种新方法。     

Immunofluorescence studies of canine distemper encephalitis on paraffin-embedded tissue.

A paraffin-embedding technique for fluorescent antibody studies of canine distemper encephalitis was developed. Specific fluorescence was demonstrated in all brain tissue from dogs with canine distemper, and when compared with tissue on cryostat sections, the paraffin-embedded tissue showed superior preservation of tissue architecture. The preservation of viral antigen was good, and the appearance of fluorescence in gray- and white-matter lesions was described. In gray matter, extensive fluorescence was found mainly in neurons; fluorescence in white matter was less extensive and was mainly associated with astrocytes.     

A catheter technique for biopsy of dogs with chronic nasal disease

A method of catheter biopsy of intranasal tissue is described. This technique was evaluated in 23 dogs referred with a history of chronic nasal disease. The technique was successful in providing diagnostically useful tissue in 15 of the cases and was shown to be a safe, inexpensive and useful diagnostic aid.     

Implementation of immunohistochemistry on frozen ear notch tissue samples in diagnosis of bovine viral diarrhea virus in persistently infected cattle

Background

Bovine viral diarrhea is a contagious disease of domestic and wild ruminants and one of the most economically important diseases in cattle. Bovine viral diarrhea virus belongs to the genus Pestivirus, within the family Flaviviridae. The identification and elimination of the persistently infected animals from herds is the initial step in the control and eradication programs. It is therefore necessary to have reliable methods for diagnosis of bovine viral diarrhea virus. One of those methods is immunohistochemistry. Immunohistochemistry on formalin fixed, paraffin embedded tissue is a routine technique in diagnosis of persistently infected cattle from ear notch tissue samples. However, such technique is inappropriate due to complicated tissue fixation process and it requires more days for preparation. On the contrary, immunohistochemistry on frozen tissue was usually applied on organs from dead animals. In this paper, for the first time, the imunohistochemistry on frozen ear notch tissue samples was described.

Findings

Seventeen ear notch tissue samples were obtained during the period 2008-2009 from persistently infected cattle. Samples were fixed in liquid nitrogen and stored on -20°C until testing. Ear notch tissue samples from all persistently infected cattle showed positive results with good section quality and possibility to determinate type of infected cells.

Conclusions

Although the number of samples was limited, this study indicated that immunohistochemistry on formalin fixed paraffin embedded tissue can be successfully replaced with immunohistochemistry on frozen ear notch tissue samples in diagnosis of persistently infected cattle.     

Pemphigus in the dog: comparison of immunofluorescence and immunoperoxidase method to demonstrate intercellular immunoglobulins in the epidermis

In 3 dogs with pemphigus vulgaris and 4 dogs with pemphigus foliaceus, intercellular immunoglobulins were demonstrated in the epidermal stratum spinosum. The immunofluorescence technique on cold ethanol-fixed and paraffin-embedded tissue sections was compared with the immunoperoxidase method on formalin-fixed paraffin-embedded tissue sections. The results of both methods were identical. However, the advantage of the unlabeled antibody-enzyme method was that the same formalin-fixed tissue specimens could be used for conventional light microscopy, as well as for immunohistologic studies.