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1.
将抗番鸭GPV单抗腹水采用透析法标记异硫氰酸荧光素(FITC),制备成抗GPV荧光抗体,研制检测GPV抗原的直接免疫荧光诊断方法。结果显示GPV荧光抗体仅与GPV阳性的组织切片或细胞呈现特异性荧光,与番鸭细小病毒(MPV)、番鸭呼肠孤病毒(MDRV)、鸭副粘病毒(DPMV)、鸭病毒性肝炎病毒(DHV)和正常番鸭组织切片不反应;与间接荧光方法的符合率为92.9%。表明GPV荧光抗体具有较好的特异性、敏感性和准确性,可用于临床快速诊断番鸭小鹅瘟病。  相似文献   

2.
根据GenBank中番鸭"三周病"病毒(MDPV)、小鹅瘟病毒(GPV)的NS基因和VP基因,以及华南农业大学禽病组分离到的细小病毒型"白点病"病毒(MDWNV)的NS和VP基因序列,设计3对特异性引物,并在建立单项PCR技术的基础上,优化多联PCR反应条件(EX-Taq DNA聚合酶浓度、引物浓度、dNTPs浓度以及退火温度等),建立了MDPV、MDWNV和GPV三种病毒的多联PCR检测方法。本研究结果表明,该番鸭三种细小病毒多联PCR检测方法具有特异、快速、准确、敏感的特点,能同时鉴别诊断这三种番鸭细小病毒,并可直接应用于临床典型病变组织的检测。  相似文献   

3.
通过对雏番鸭细小病毒(MDPV)高免血清与来自江苏、广东两系小鹅瘟细小病毒(GPV)高免血清和小鸭病毒性肝炎(DHV)高免血清的比较检测证明,微量碘凝集试验(MIAT)对MDPV高免血清具有特异性。用于检测实验室人工感染和野外自然感染痊愈病例血清,均呈阳性反应;而GPV高免血清和DHV高免血清及健康番鸭正常血清均无此反应。因此,MIAT可作为检测雏番鸭细小病毒的一种简便、特异、快速和敏感的诊断法。  相似文献   

4.
应用GPV胶乳凝集方法(LPA)和病毒分离(VI)检测人工感染病例9份、健康对照6份,临床疑似送检病鸭28份。其中人工感染样品检测LPA和VI符合率为93.3%(14/15);临床疑似样品检测LPA和VI符合率为89.3%(25/28);两者总符合率达90.7%(39/43)。LPA方法具有简便(肉眼可判定,不需仪器)、快速(检测抗原20min内)、特异、准确等优点,比VI更适合临床使用。  相似文献   

5.
为建立鉴别鸭瘟病毒(DPV)、鹅细小病毒(GPV)和番鸭细小病毒(MDPV)的多重PCR检测方法,参考文献合成针对DPVUL6基因、GPVNS基因和MDPVVP1基因序列的特异性引物。通过优化多重PCR反应中的引物浓度和退火温度,建立快速鉴别3种病毒的多重PCR方法,并对该方法的特异性、敏感性和重复性进行检验。结果显示,该方法对禽流感病毒、新城疫病毒、鸭肝炎病毒、鸭源巴氏杆菌、鸭疫里氏杆菌的核酸扩增结果均为阴性,表明特异性较强;DPV、GPV和MDPV的最低核酸检出量分别为237.3pg、22.45pg和204.6pg,表明敏感性良好;该方法对DPV+GPV+MDPV、DPV+GPV、DPV+MDPV、GPV+MDPV、DPV、GPV和MDPV的核酸均能扩增出与预期大小一致的目的条带,表明该多重PCR方法重复性较好。本研究方法具有特异性强、敏感性良好、重复性较好和快速简便等特点,可用于DPV、GPV和MDPV临床感染病例的联合检测与鉴别诊断。  相似文献   

6.
依据GenBank小鹅瘟病毒(GPV)的NS基因保守区设计一对特异性引物,利用PCR扩增NS区片段,克隆、测序,采取Taqman探针法,构建小鹅瘟病毒的荧光PCR检测方法.结果表明,该方法检测GPV具有很高的特异性和敏感性,可以用于小鹅瘟病毒的检测.  相似文献   

7.
为建立一种敏感、自动化,高通量的检测病料中鹅细小病毒(GPV)的PCR-DHPLC检测方法,本研究首先根据GenBank上GPV标准毒株VP3基因序列设计一对引物,建立检测GPV的PCR方法,然后将PCR检测方法与DHPLC法进行有机结合,建立了GPV PCR-DHPLC检测方法。以GPV、鹅副粘病毒、鸭瘟病毒及正常番鸭胚尿囊液进行特异性试验,结果无交叉反应,表明该检测方法具有较高的特异性。对GPV阳性样品检测结果表明该方法的敏感性较高,是常规PCR电泳法的100倍,且具有较好的重复性。对临床上采集的100份疑似病料,分别应用PCR电泳法和本试验建立的PCR-DHPLC方法进行检测,结果阳性符合率为100%。表明该方法具有特异、敏感、快速、重复性好、可高通量检测等优点,可用于临床样品的大批量检测。  相似文献   

8.
为建立水禽源细小病毒的快速检测方法,本研究采用纯化的小鹅瘟病毒(GPV)免疫BALB/c小鼠,取免疫小鼠脾细胞与SP2/0骨髓瘤细胞进行融合,筛选到21株能够稳定分泌抗GPV的单克隆抗体(MAb),将1株MAb纯化后与胶体金结合,喷涂金标垫,与另一株纯化的MAb配对,制成检测水禽源细小病毒的胶体金试纸条。特异性试验表明,鹅细小病毒、番鸭细小病毒和新型番鸭细小病毒的检测结果均为阳性,猪细小病毒、1型鸭甲肝病毒、坦布苏病毒、产蛋下降综合征病毒、H9N2亚型禽流感病毒、番鸭呼肠孤病毒和新城疫病毒等的检测结果均为阴性。该方法对鹅细小病毒、番鸭细小病毒和新型番鸭细小病毒的最低检出量分别为10~(4.7) ELD50/mL、10~(4.7)ELD50/mL和10~(5.7) ELD50/mL。重复性试验良好。对临床粪便样品的检测,该方法比PCR方法简单快速,但是存在漏检的情况。上述结果表明,本实验建立的方法可用于水禽源细小病毒病的初步快速筛查。  相似文献   

9.
采用PCR方法扩增GPV VP1基因173 bp的基因片段,克隆到PMD18-T载体上,将纯化后的质粒做为模板,建立扩增反应的标准曲线,最终建立了小鹅瘟病毒(GPV)SYBR GreenⅠ荧光定量PCR检测方法。该方法检测灵敏性高,可检测2×101拷贝/μL的模板量;特异性好,与鹅副黏病毒、鹅流感病毒、鹅源鸭瘟病毒、大肠杆菌、沙门氏菌、曲霉菌均不发生交叉反应。建立的GPV荧光定量PCR具有特异性好、敏感度高、检测时间短、结果重复性好等优点,可用于小鹅瘟的临床诊断和流行病学调查。  相似文献   

10.
根据GenBank登录的鹅细小病毒(GPV)和番鸭细小病毒(MDPV)非结构蛋白(NS)基因特征,本研究设计1对特异性引物对GPV和MDPV基因组DNA进行PCR扩增,目的片段大小均为810 bp,并对PCR产物进行切胶回收。用EcoRⅠ酶对GPV和MDPV特异性胶回收产物进行酶切鉴定,结果显示MDPV经EcoRⅠ酶切后琼脂糖凝胶电泳检测片段为2段,大小为530和280 bp;而GPV经EcoRⅠ酶切后琼脂糖凝胶电泳检测片段大小不变。本研究建立了一种快速区别GPV和MDPV感染的检测方法,可对番鸭感染水禽细小病毒的情况进行快速鉴别诊断。  相似文献   

11.
12.
在现代法律秩序中,商会自治规范是制定法的基础和必要的补充,甚至在某些方面替代了制定法;商会自治规范主要包括商会组织规范、行为规范、惩罚规范以及争端解决规范等;其效力仅及于其内部成员;商会自治规范和制定法之间存在冲突,但也存在整合的基础。  相似文献   

13.
采用高效液相色谱法测定癸氧喹酯干混悬剂的含量,在2-250μg/mL范围内,峰面积的常用对数与进样量浓度的常用对数呈良好的线性关系,R^2=1(n=5),平均回收率为99.24%~99.51%,RSD在0.05%~0.28%。此方法分析时间短,样品前处理简便、定量结果准确,重现性好,结果满意,为其质量控制提供了依据。  相似文献   

14.
本文概述了猪的毛色类型、猪的毛色遗传模式,着重综述了猪毛色基因分子基础的研究进展,指出存在问题并就未来发展方向做了思考。  相似文献   

15.
REASONS FOR PERFORMING STUDY: Centesis of the bicipital bursa using an 8.9 cm long spinal needle has been reported but the alternative of employing a 3.8 cm long hypodermic needle requires validation. OBJECTIVE: To compare the efficacy of 2 different methods of centesis of the bicipital bursa and to evaluate the usefulness of ultrasonographic imaging to determine the location of solution administered when centesis of the bursa is attempted. METHODS: For Trial 1, 6 clinicians, who had no previous experience of centesis of the bicipital bursa, attempted to inject a solution composed of an aqueous radiopaque contrast medium and physiological saline solution (PSS) into the bicipital bursae of 2/12 horses using the previously described distal approach to inject one bursa and a proximal approach to inject the contralateral bursa. The bicipital tendon and bursa were examined ultrasonographically before and after injection; and both shoulders were examined radiographically to identify the location of the medium. In Trial 2, another 6 clinicians, also with no previous experience of centesis, repeated Trial 1, using 6 horses, but the radiopaque contrast medium was mixed with air instead of PSS. RESULTS: Accuracy of centesis using the proximal approach was 39% and that of the distal approach 28%. Ultrasonographic examination of the shoulder allowed the location of solution and air to be accurately predicted in all 12 shoulders examined. CONCLUSIONS: Clinicians who have had no previous experience performing centesis of the bicipital bursa are unlikely to be successful in centesis using either approach. Radiographic examination after injecting a radiopaque contrast medium may be necessary to assess the success of centesis especially if bursal fluid is not obtained during centesis. Injecting air along with the radiopaque contrast medium provides more accurate ultrasonographic confirmation of centesis and better radiographic definition than does injection without air.  相似文献   

16.
用硝酸和高氯酸消化蜂蜜,使硒游离出来,在微酸性环境下,硒和2,3-二氨基萘(DAN)生成有较强荧光的物质,用环己烷萃取,在激发波长378nm,荧光波长518nm处测定其荧光强度。蜂蜜中硒含量范围:0.10~0.82μg/g。表明:蜂蜜应视为天然富硒营养品。  相似文献   

17.
乳酸杆菌益生作用机制的研究进展   总被引:2,自引:0,他引:2  
乳酸杆菌作为益生菌广泛用于人和动物。本文综述了乳酸杆菌改善宿主健康的机制。乳酸杆菌可通过产生抗菌物质如乳酸、过氧化氢、细菌素,或者通过竞争营养或肠道黏附位点来抑制致病菌;通过诱导黏附素的分泌或阻止细胞凋亡而增强肠道的屏障功能,从而保护肠道。文章重点讨论了乳酸杆菌表面成分(表面蛋白、脂磷壁酸和肽聚糖)与肠道受体(C型凝集素受体、Toll样受体和 Nod样受体),阐述了他们结合后启动免疫调节信号,调控肠道免疫功能以发挥改善健康作用的机制。  相似文献   

18.
Ingestively masticated fragments were collected and sized via sieving. Different sizes of esophageal masticate and ruminal digesta fragments, and ground fragments of larger masticated pieces were incubated in vitro, and undigested NDF remaining at intervals of up to 168 h of incubation was determined. The ruminal age-dependent time delay (tau) for onset of digestion of NDF was positively correlated (P < 0.004) with the mean sieve aperture estimated to retain 50% of the fragments between successive sieve apertures (MRA). Degradation rate of potentially degradable NDF (PDF) and level of indigestible NDF were not related (P > 0.10) to MRA of masticated and ground fragments. Estimates of tau were positively related to MRA, with slopes of bermudagrass < corn silage < ruminal fragments of corn silage. It was concluded that fragment size-, and consequently, ruminal age-dependent onset of PDF degradation of a mixture of different fragment sizes results in an age-dependent rate of degradation of the more rapidly degrading of two subentities of PDF. Models are proposed that assume a tau before onset of simultaneous degradation of PDF from two pools characterized as having gamma-modeled age-dependency and age-constant rates. The ruminal age-dependent pool seems to be associated with the faster-degrading pool, and its rate parameter increases with range in MRA in the population of fragments. Conceptually, the ruminal age-dependent rate parameter for PDF degradation seems to represent a composite of several effects: 1) effects of the size-dependent tau; 2) range in MRA of the population of ingestively masticated fragments; and 3) subentities of PDF that degrade via more rapid age-dependent rates compared with subentities of PDF that degrade via age-constant rates. The estimated fractional rates of ruminative comminution of ingestively masticated fragments (0.060 to 0.075/h) were of a magnitude similar to the mean fractional rates of PDF digestion (0.030 to 0.085/h), which implies that ruminative comminution may be first-limiting to fractional rate of PDF digestion. The in vivo roles of ingestive and ruminative mastication of fragments on PDF degradation must be considered in any kinetic system for estimating PDF digestion in the rumen. These results and others in the literature suggest that the rate of surface area exposure rather than intrinsic chemical attributes of PDF may be first-limiting to degradation rate of PDF in vivo.  相似文献   

19.
乳酸杆菌作为益生菌广泛用于人和动物.本文综述了乳酸杆菌改善宿主健康的机制.乳酸杆菌可通过产生抗菌物质如乳酸、过氧化氢、细菌素,或者通过竞争营养或肠道黏附位点来抑制致病菌;通过诱导黏附素的分泌或阻止细胞凋亡而增强肠道的屏障功能,从而保护肠道.文章重点讨论了乳酸杆菌表面成分(表面蛋白、脂磷壁酸和肽聚糖)与肠道受体(C型凝集素受体、Toll样受体和Nod样受体),阐述了他们结合后启动免疫调节信号,调控肠道免疫功能以发挥改善健康作用的机制.  相似文献   

20.
OBJECTIVE: To determine whether serum concentrations of biomarkers of skeletal metabolism can, in conjunction with radiographic evaluation, indicate severity of osteochondrosis in developing horses. ANIMALS: 43 Dutch Warmblood foals with varying severity of osteochondrosis. PROCEDURE: 24 foals were monitored for 5 months and 19 foals were monitored for 11 months. Monthly radiographs of femoropatellar-femorotibial and tibio-tarsal joints were graded for osteochondral abnormalities. Serial blood samples were assayed for 8 cartilage and bone biomarkers. At the end of the monitoring period, foals were examined for macroscopic osteochondrosis lesions. RESULTS: Temporal relationships were evident between certain serum biomarkers and osteochondrosis severity in foals during their first year. Biomarkers of collagen degradation (collagenase-generated neoepitopes of type-II collagen fragments, type-I and -II collagen fragments [COL2-3/4C(short)], and cross-linked telopeptide fragments of type-I collagen) and bone mineralization (osteocalcin) were positive indicators of osteochondrosis severity at 5 months of age. In foals with lesions at 11 months of age, osteochondrosis severity correlated negatively with COL2-3/4C(short) and osteocalcin and positively with C-propeptide of type-II procollagen (CPII), a collagen synthesis marker. Radiographic grading of osteochondrosis lesions significantly correlated with macroscopic osteochondrosis severity score at both ages and was strongest when combined with osteocalcin at 5 months and CPII at 11 months. CONCLUSIONS AND CLINICAL RELEVANCE: The ability of serum biomarkers to indicate osteochondrosis severity appears to depend on stage of disease and is strengthened with radiography. In older foals with more permanent lesions, osteochondrosis severity is significantly related to biomarker concentrations of decreased bone formation and increased cartilage synthesis.  相似文献   

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