美味侧耳五种酶的同工酶谱在不同菌体组织中的变化

比较了美味侧耳不同菌体组织中５种酶的同工酶谱变化。酯酶（Ｅ５Ｔ）和苹果酸脱氢酶（ＭＤＨ）在子实体中的酶带数和活性均高于液培菌丝。过氧化物酶（ＰＯＤ）的情况则正好相反。乙醇脱氢酶（ＡＤＨ）和山梨醇脱氢酶（ＳＯＤＨ）的酶带只在子实体中发现。酶谱随子实体部位而变异的进度同酶种类有关，ＰＯＤ变化较大，ＥＳＴ和ＭＤＨ较不明显，ＡＤＨ和ＳＯＤＨ没有变化。本实验结果为今后以这５种酶为生化遗传标记研究美味侧耳提供了基础依据。     

Sensitivity,specificity, and predictive values of reagent test strip estimations of blood urea nitrogen

Azostix-reagent-tests(R) strips (Ames, Miles, Inc., Diagnostic Division, Elkhart, IN) were used to measure blood urea nitrogen values in blood samples from 125 dogs and cats at the North Carolina State University, College of Veterinary Medicine. Results of the tests were compared with standard serum urea nitrogen results. Sensitivity, specificity, and negative predictive values were high (86.4, 90.3, and 96.5%, respectively). Positive predictive value was low, 65.5% of the dogs and cats with elevated blood urea nitrogen values were correctly classified as abnormal The test performs well when the prevalence of abnormal values is near 50%.     

Validation of a multiplex PCR assay to detect Babesia spp. and Anaplasma marginale in cattle in Uruguay in the absence of a gold standard test

Detection of bovine Babesia spp. and Anaplasma marginale is based on the reading of Giemsa-stained blood or organ smears, which can have low sensitivity. Our aim was to improve the detection of bovine Babesia spp. and A. marginale by validating a multiplex PCR (mPCR). We used 466 samples of blood and/or organs of animals with signs and presumptive autopsy findings of babesiosis or anaplasmosis. The primers in our mPCR amplified the rap-1a gene region of Babesia bovis and B. bigemina, and the msp-5 region of A. marginale. We used a Bayesian model with a non-informative priori distribution for the prevalence estimate and informative priori distribution for estimation of sensitivity and specificity. The sensitivity and specificity for smear detection of Babesia spp. were 68.6% and 99.1%, and for A. marginale 85.6% and 98.8%, respectively. Sensitivity and specificity for mPCR detection for Babesia spp. were 94.2% and 97.1%, and for A. marginale 95.2% and 92.7%, respectively. Our mPCR had good accuracy in detecting Babesia spp. and A. marginale, and would be a reliable test for veterinarians to choose the correct treatment for each agent.     

花鲈虹彩病毒交叉引物恒温扩增检测方法的建立

【目的】花鲈虹彩病毒(Lateolabrax maculatus iridovirus,LMIV)严重威胁花鲈养殖业安全,无特效防控药物,早期诊断在LMIV防控中发挥极其重要的作用。建立一种简便、快捷、准确的现场快速诊断方法,可为LMIV的基层诊断提供技术支撑。【方法】利用交叉引物恒温扩增技术(Cross priming amplification,CPA),针对LMIV ATPase基因高保守区设计1套单交叉引物。以构建的ATPase重组质粒作为阳性模板,对反应体系中的引物浓度比,Bst DNA聚合酶、Betaine、MgSO₄、dNTP浓度,以及反应温度和反应时间进行优化;结合一次性核酸试纸条,建立可视化检测LMIV-CPA的方法。【结果】最优引物浓度比组合为交叉引物CPF1.0μmol/L,引物F3和B3均为0.4μmol/L,探针引物B1(FAM)和B2(Biotin)均为0.8μmol/L;MgSO4浓度为6 mmol/L、Betaine浓度为0.4 mol/L、dNTP浓度为0.6 mmol/L、Bst DNA聚合酶浓度为0.256 U/μL;最佳反应...     

弧菌属PCR检测方法的建立与应用

根据弧菌属细菌(Vibrio spp.)共有的rpo A基因序列,比对和设计了一对PCR引物,建立了能够快速而准确地检测弧菌属细菌的通用PCR检测方法,该方法对靶标DNA的检测灵敏度为58 fg/μL,对菌液的检测灵敏度为2.7×102cfu/m L,能够区分该属细菌与其它属的细菌,具有极高的特异性。使用建立的方法对分离自南美白对虾的病原菌进行了检验,并结合16S r DNA序列分析,结果显示待检测病原均为弧菌属的细菌,与测序结果一致,说明该检测方法可用于弧菌属细菌的检测和诊断。     

基于牡蛎疱疹病毒DNA聚合酶基因的巢式PCR检测方法的建立及应用

为了建立适用于Os HV-1不同变异株的检测方法,在牡蛎疱疹病毒(Os HV-1)3个变异株全基因组序列比对的基础上,筛选到牡蛎疱疹病毒基因组中高度保守的DNA聚合酶(DNA polymerase)基因,据此设计巢式PCR引物,优化PCR反应体系和条件,建立了基于Os HV-1 DNA聚合酶基因的巢氏PCR检测方法(P-n PCR检测方法),利用P-n PCR与Cn PCR检测方法对不同年份和宿主来源的Os HV-1疑似感染样本进行检测。结果显示,Pn PCR检测方法能稳定地检出100拷贝/μL的病毒DNA;P-n PCR较C-n PCR检测方法具有更强的特异性和更高的检出率。研究表明,本研究建立的P-n PCR检测方法适用于Os HV-1不同变异株的检测,可为该病毒的检测和流行病学调查提供可靠的技术支持。     

金银花多酚氧化酶特异性与抑制剂动力学研究

对金银花多酚氧化酶底物特异性和不同抑制剂的抑制效应进行了动力学分析,结果表明:金银花多酚氧化酶的最适作用底物为绿原酸,Km值为0.0059 mmol/L。维生素C、4-己基间苯二酚、L-半胱氨酸对多酚氧化酶的抑制均属可逆抑制,其抑制类型分别属于混合性、竞争性、反竞争性,对游离酶的抑制常数KI分别为1.620、4.587、0 mmol/L,对酶-底物络合物的抑制常数KIS分别为1.995、0、3.780 mmol/L;柠檬酸的抑制效应属不可逆抑制。维生素C、4-己基间苯二酚、L-半胱氨酸、柠檬酸对金银花多酚氧化酶半抑制浓度分别为0.062、0.053、0.140、0.048 mmol/L。     

豚草(Ambrosia artemisiifolia)不同器官与不同发育期中过氧化物酶同工酶分析

本文用凝胶电泳法测定了豚草根、茎、叶不同器官过氧化物酶同工酶酶谱,按其酶带相对迁移率(R_m),可分为11条酶带.豚草植株过氧化物酶同工酶与其他植物一样,有明显的器官特异性和阶段发育特异性.结果表明豚草茎叶中过氧化物酶酶谱在开花前后有明显的变化:(1)开花期较开花前酶带明显地增多;(2)开花期在主茎或叶片中,都会出现一条颜色较浅的,该器官花期特有的酶带;(3)豚草株体内一条主酶带(E 带)在开花期明显变浅.上述过氧化物酶同工酶的三种变化,为判断豚草植株体内开花前后生理生化变化提供了一种生化指标.