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为建立一种快速、特异、灵敏的检测鸡细小病毒(chicken parvovirus,ChPV)的方法,根据ChPV的保守基因NS1设计了3条特异性引物,建立并优化了能快速检测ChPV的半巢式PCR方法,对其进行特异性和敏感性试验,并用所建立的方法对48份临床样品进行了检测。特异性和敏感性试验结果显示,建立的半巢式PCR只对ChPV敏感,扩增产物为186 bp的特异性条带;其最低能检测到5.62 fg/μL的ChPV DNA;而对鸡新城疫病毒、H9亚型禽流感病毒、马立克氏病病毒、鸡传染性喉气管炎病毒、鸡传染性支气管炎病毒不敏感。临床检测结果显示,同时对48份临床样品进行检测,检出率为16.67%(8/48),提示广西区内鸡群存在ChPV感染。本研究建立的ChPV半巢式PCR方法适用于ChPV的临床检测。 相似文献
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建立可同时检测鸡细小病毒(Chicken parvovirus,ChPV)与禽呼肠病毒(Avian reovirus,ARV)的二重PCR方法,为防控ChPV与ARV提供技术支撑。根据鸡细小病毒NS1基因和禽呼肠病毒σC基因的保守序列,设计合成两对引物用于检测ChPV和ARV,通过优化二重PCR的反应体系,特异性、敏感性试验评价建立的ChPV与ARV二重PCR。优化后的二重PCR反应体系为:2×PCR Mix 12.5μL,其中ChPV与ARV的上、下游引物各1.0μL,混合模板2.0μL,ddH2O补足25μL;最佳的反应程序为:95℃5min;95℃1min,56.1℃1min,72℃1min,35个循环;最后72℃延伸10min。结果显示,建立的二重PCR能够同时扩增出204bp ChPV和405bp ARV片段;该方法对ChPV与ARV的检测敏感性分别达到58fg和53fg,但对鸡新城疫病毒、H9亚型禽流感病毒、马立克病病毒、鸡传染性喉气管炎病毒、鸡传染性支气管炎病毒等病原体均无特异性扩增,对ChPV与ARV混合感染的临床阳性病料的检测结果与各病毒单项PCR检测结果符合率为94%以上。建立的二重PCR可用于ChPV与ARV感染的快速鉴别诊断。 相似文献
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为了同时检测禽肾炎病毒(avian nephritis virus,ANV)和鸡细小病毒(chicken parvovirus,ChPV),根据GenBank中ANV的ORF基因和ChPV的NS1基因的保守序列,设计筛选了扩增片段大小分别为511 bp和242 bp的特异性引物,通过反应条件优化、特异性和敏感性试验,建立了一种双重PCR检测方法。该方法最佳引物比例为ANV上下引物各0.4μL(20 mol/L),ChPV上下引物各0.6μL(20 mol/L);最佳退火温度为53.8℃;灵敏度可达到55 fg(ANV)和58 fg(ChPV);对常见鸡病病原体进行检测,结果全为阴性。本研究建立的PCR方法具有特异、敏感、快速、稳定的优点,可用于同时鉴别诊断ANV和ChPV的混合感染。 相似文献
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为建立一种能同时鉴别诊断禽流感病毒(avian influenza virus,AIV)和鸡细小病毒(chicken parvovirus,ChPV)的检测方法,本研究根据GenBank中AIV的M基因和ChPV的NS基因保守序列,分别设计并筛选出两对特异性引物,用于AIV和ChPV的检测。通过优化反应条件,建立了AIV和ChPV二重PCR检测方法。试验结果表明,该方法特异好,能同时检测AIV和ChPV,对其他常见的禽病病原体均未反应;该法对AIV和ChPV的检测下限均为100 fg;对159份临床样品检测结果与PCR阳性产物测序结果一致。本研究建立的AIV和ChPV二重PCR检测方法具有特异性好、灵敏度高的特点,对AIV和ChPV的防制具有重要意义。 相似文献
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旨在建立一种可定量检测鸡细小病毒(Chicken parvovirus, ChPV)的微滴式数字PCR(droplet digital PCR,dd PCR)新型检测方法。引物设计参考了GenBank中ChPV的NS基因保守序列,筛选了特异性探针引物组,采用不同探针引物浓度组合和退火温度梯度对反应条件进行优化,并通过使用该方法与荧光定量PCR(qPCR)和普通PCR方法进行灵敏度比较,同时对其他常见禽病病原体进行检测,以及对40份临床样品进行检测,评估了该方法的特异性、敏感性和重复性。结果显示,该方法最佳引物浓度均为900 nmol/L,最佳探针浓度为600 nmol/L;最佳退火温度为54.0℃;灵敏度比qPCR和普通PCR分别高10倍和1000倍,最低检测限为4.5 copies/μL;对其他常见禽病病原体进行ChPV微滴式数字PCR检测,结果均为阴性;批内和批间重复性较好,变异系数小。结果表明,所建立的ddPCR方法具有更高的敏感性和准确性,可成功用于快速定量检测ChPV感染。 相似文献
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为探明2021年龙岩市规模化鸡场鸡传染性贫血病病毒(CIAV)、圆圈病毒3型(GyV3)、鸡细小病毒(ChPV)、J亚群禽白血病病毒(ALV-J)等4种病毒的感染情况,对龙岩市32个规模化鸡场采集的297份病死鸡组织样品进行PCR或RT-PCR检测,并对检测结果进行统计分析.结果显示:CIAV、GyV3、ChPV和AL... 相似文献
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应用PCR方法检测鸡传染性贫血病毒 总被引:3,自引:0,他引:3
根据发表的鸡传染性贫血病毒(CAV)序列,选择鸡传染性贫血病毒基因组保守区域。设计一对引物,建立了CAV快速诊断的PCR方法。特异性结果说明,该方法对CAV不同毒株均能扩增出明显的特异性条带,而新城疫病毒(NDV)Lasota株、传染性支气管炎病毒(IBV)M41株、鸡马立克氏病病毒(MDV)和鸡传染性法氏囊病病毒(IBDV)扩增结果均为阴性;PCR反应敏感性结果显示.该方法能检测出的DNA最低浓度为O.001pg/μl,可用于临床感染不同年龄鸡群的CAV的检测。 相似文献
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魏静 《四川畜牧兽医学院学报》2009,(4):28-32
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
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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
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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
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以国际标准强毒R株人工感染非免疫产蛋鸡,定时扑杀,分别从鼻窦、眶下孔、气管、肺、气囊、卵巢和输卵管分离MG,并收集感染鸡所产蛋分离MG。结果表明,人工感染48小时后上、下呼吸道及肺已被全面感染,96小时气囊已被感染,120小时输卵管已能分离到MG,卵巢始终分离不到MG。人工感染鸡自144小时便能在其所产蛋中分离出MG。药物治疗能在72小时内消除感染,油乳剂苗则需24天后逐渐降低蛋内MG分离率,药物卵内注射、种蛋药浴、高温处理均能杀死卵内MG,但以研制的种蛋浸泡剂药浴效果为最好。 相似文献
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Effects of size of ingestively masticated fragments of plant tissues on kinetics of digestion of NDF
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