PRRSVNsp9蛋白基因的RT—LAMP检测方法的建立

根据GenBank中登录的PRRSVNsp9基因序列,应用PrimerExplorerV4软件,在该基因序列中选取保守区设计了4条RT—LAMP引物,旨在建立1种针对Nsp9蛋白基因的逆转录环介导等温扩增（RT-LAMP）的快速检测方法。采用引物分组扩增的方法对引物进行了检测,以确保引物的可靠性。对反应体系、温度以及时间进行了优化,检测了该方法的特异性和灵敏度。结果表明,该方法在等温条件下只需50min就能检测出结果。与RT-PCR方法相比,在判定检测结果时不需要借助昂贵仪器设备,具有高特异性、高灵敏度、操作简便快速等特点,适合于临床检测的应用。     

犬巴贝斯虫病检测研究

犬巴贝斯虫病是经蜱传播的血液寄生虫病,其临床症状是高热、贫血、血红蛋白尿和脾肿大。通过血涂片检验,血常规检验并结合临床特征,对西安市103只宠物犬进行了巴贝斯虫的感染情况检测。结果显示,犬血液中巴贝斯虫的感染数为16例,占总数的15.53%,其中5月份的感染犬最多,为13例,占感染数的81.25%(13/16)。     

Research Progress on Rapid Serological Assays for Diagnosis of Japanese Encephalitis B

Japanese encephalitis B (JE) is a zoonosis caused by Japanese encephalitis virus which invades the central nervous system,infected by mosquito.Various animals could be the sower and reservoir of JE including swine,horse,cattle and avian.JE could cause a major public health trouble and a horrible crisis of the farm all in Asia and the Western Pacific region.Therefore,it is necessary to diagnose it rapidly and accurately.This review article made a sketch of approaches for the rapid serological assays of JE,including traditional method,emerging technologies and specific improvements of the assay,so as to provide references for the study of rapid,simple and accurate JE diagnose methods.     

广西不同年龄猪群中2型猪链球菌抗体水平调查分析

为调查规模化猪场不同年龄猪群中2型猪链球菌(S.suis 2)抗体水平,本研究采用以荚膜多糖为抗原的间接ELISA对来自广西地区母猪群、肥猪群、生长猪群共1 908份血清样品进行了S.suis 2抗体检测和分析。结果显示:成年猪群中抗S.suis 2的抗体水平普遍较高,种猪场母猪血清抗体阳性率可高达95%以上,不同地区屠宰场的育肥猪血清抗体阳性率在39.2%~97.5%。对不同日龄仔猪群抗体水平检测分析显示:14日龄~42日龄期间,抗体水平随仔猪日龄的增长而呈逐渐下降的趋势;而42日龄~130日龄期间,抗体水平随日龄的增长而呈逐渐上升的趋势。此外,经产母猪群抗体水平略高于后备母猪群,并且随胎次的增加,抗体的离散度缩小。以上结果表明S.suis 2抗体在仔猪生长阶段的空白期即42日龄时,仔猪群抗体水平最低。因此,在这个阶段和之前做好S.suis的预防和免疫工作对猪群链球菌病的防控至关重要。     

耐氟喹诺酮类药物大肠埃希氏菌基因突变的MAMA-PCR检测

采用MAMA-PCR法对72株已知突变菌株的突变情况进行了检测。结果，有2种gyrA83位突变：TCG→TTG和TCG缺失；4种gyrA87位突变：GAC→AAC、GAC→GGC、GAC→TAC和GAC→CAC；1种parC80位突变：AGC→ATC以及4种parC84位突变：GAA→AAA、GAA→GCA、GAA→GGA和GAA→GTA。采用同样的方法，对38株未知突变菌株的检测结果显示，gyrA83、gyrA87、parC 80和parC84位的突变检出的正确率分别为97．4％、94．7％、81．6％和94．7％。结果表明，MAMA-PCR有望成为监测耐氟喹诺酮类药物大肠埃希氏菌的分子生物学常规方法。     

广州桑树植原体分子检测及多样性初探

采用植原体16S-23S rDNA区段的通用引物对P1/P7和巢式引物对Rm16F2/Rm16R1,建立了快速准确的桑树植原体巢式PCR检测技术。对广州的两个桑树品种资源圃中的部分桑树品种进行了植原体分子检测,结果在两个资源圃中均发现有植原体存在。对巢式PCR的扩增产物(16S rDNA片段)进行了限制性片断长度多态性(RFLP)分析,显示出3种RFLP带型,暗示桑树植原体存在多样性。对所得植原体16S rDNA片段进行序列测定,并与其它植物植原体作亲缘关系分析,结果表明该植原体的16S rDNA序列与其它植物病原植原体之间的同源性为83.3%~99.9%,并初步判断所检测到的桑树植原体属于16S rI组。     

上海市规模化猪场保育猪传染病的调查分析

于2003年-2004年间对上海的13个规模化猪场的保育猪进行了集中采样,并结合临床检查、细菌学检测、血清学检测和病毒学诊断等方式对保育猪传染病进行了综合性调查,基本弄清了保育猪传染病的主要致病因子,为采取切实有效的针对性措施提供了技术支持。     

链霉素胶体金快速检测试纸条的研制

本研究建立了一种基于免疫胶体金技术的链霉素快速检测方法。该方法检测灵敏度10ng·ml-(101ppb),蜂蜜样本检出限是40ng·ml-(40ppb),单个样本检测时间为5～10min。所研制试纸条产品检测变异系数小,批1间、批内检测结果重复性良好,可以应用于蜂蜜样本中链霉素的快速检测和现场筛查,具有较高的市场应用前景。     

两种热带森林覆盖变化检测方法比较研究

[目的]以热带森林复杂区域为对象,对两种缨帽变换“衍生数据”检测热带森林变化的方法的优劣进行比较.[方法]对两期数据进行缨帽变换,同时结合黑暗对象掩膜与局部直方图阈值提取等方法,获取亮度、绿度、湿度指数组合(MKT)和干扰指数(DI).采用MKT差值(MKT-D)、干扰指数差值(MDI-D)进行变化信息识别,然后,根据植被覆盖与亮度、绿度、湿度之间的变化关系,通过决策树分类提取变化信息,最后,对不同检测结果进行验证与比较分析.[结果]结果显示两种方法都能检测出森林内部的细微变化,但对小图斑变化,MKT-D检测优势明显,且MKT-D的kappa系数为0.763 0,MDI-D的kappa系数为0.655 9,两者相比,MKT-D方法优于MDI-D方法.[结论]MKT能够增强短波红外与近红外波段对森林变化信息的敏感性,有效地消除噪声等非目标信号,突出目标信号,此外,MKT-D为RGB彩色图像,更利于变化信息的目标提取与解译.     

Canine Haematopoietic Chimerism Analyses by Semiquantitative Fluorescence Detection of Variable Number of Tandem Repeat Polymorphism

Canine models are successfully applied to the study of haematopoietic stem cell transplantation (HSCT). Monitoring of haematopoietic donor/recipient chimerism is of major significance in detecting and quantifying engraftment or graft rejection of the donor-derived haematopoietic cells after transplantation. Radioactive analyses of polymorphic microsatellite markers are commonly used for chimerism analyses. We describe an improved, non-isotopic method that is based on the analysis of microsatellite markers in donor and recipient cells using capillary electrophoresis and fluorescence detection. Artificial mixtures of donor and recipient DNA that were generated from peripheral blood mononuclear cells from dog leukocyte antigen-identical siblings were used to analyse the sensitivity of the assay. DNA from dogs that had received HSCT were also analysed in order to demonstrate the feasibility of the method in vivo. For chimerism analyses, six different microsatellite loci were systematically amplified using fluorescent PCR primer. The fluorescent polymerase chain reaction products were separated by capillary electrophoresis using POP4 on a 310 ABI Prism Genetic Analyzer. After electrophoresis, fluorescence signals were automatically sized and quantified using GeneScan software. The method described provides an accurate assessment of haematopoietic chimerism in the canine model with significantly reduced hands-on time compared to conventional gel electrophoresis.