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以胸膜肺炎放线杆菌(App)apx ⅣA基因和猪圆环病毒2型(PCV-2)ORF2基因为靶基因,分别设计特异性引物和探针,建立App和PCV-2的二重荧光PCR方法。App引物扩增的目的片段大小为132bp,PCV-2为169bp。建立的方法可在同一反应体系中同时对App和PCV-2进行定量检测鉴别,其敏感性分别为1×101拷贝和1×100拷贝,对其他猪病病原核酸的检测为阴性。利用该方法对10份人工感染临床样品进行检测试验,其结果符合率达100%,说明该方法可用于对临床样品中App和PCV-2的检测。 相似文献
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猪胸膜肺炎放线杆菌PCR诊断方法的建立与应用 总被引：5，自引：3，他引：5
根据胸膜肺炎放线杆菌ApxⅣ基因设计1对引物，扩增特异的650bp棱酸片段，建立了应用PCR检测猪胸膜肺炎放线杆菌的方法。特异性试验结果表明，12个血清型的放线杆菌参考菌株均能扩增出650bp特异性的核酸片段，而大肠杆菌、多杀性巴氏杆菌、猪肺炎支原体、伤寒沙门氏菌和支气管败血性波氏杆菌的扩增结果均为阴性。敏感性试验结果表明，PCR的最低检出限量为500个放线杆菌。利用建立的PCR检测方法对22株从山东省不同地区分离的疑似胸膜肺炎放线杆菌菌株进行检测．结果14株为阳性。对感染猪病变组织的检测结果表明，病变部位不同，胸膜肺炎放线杆菌的检出率不同，其中以扁桃体的检出率最高。 相似文献
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根据已经发表的猪传染性胸膜肺炎放线杆菌(App)dsbE-like基因的序列,设计了一对可扩增374bp目的片段的特异性引物。经PCR扩增,App1-4和6-10型标准菌株均能扩出约374bp的片段,而对大肠埃希菌(K88)、巴氏杆菌(5:A)和链球菌(Ⅱ)的扩增结果均为阴性。用建立的方法来检测分离菌株MC、ME和MH,结果表明,该方法能用于临床分离株的检测。灵敏性试验表明本方法对App菌液最低检出浓度为71cfu/mL。建立的PCR检测App的方法可用于临床上对App的检测。 相似文献
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猪的胸膜肺炎放线杆菌、多杀性巴氏杆菌和副猪嗜血杆菌复合PCR检测方法的建立 总被引：9，自引：0，他引：9
在已经建立猪胸膜肺炎放线杆菌(APP)、猪多杀性巴氏杆菌(PM)、副猪嗜血杆菌(HPS)的单项PCR诊断方法的基础上,通过对扩增条件的优化,成功地建立了APP、PM、HPS的复合PCR实验室诊断方法,利用一次PCR反应,即可同时扩增出APP的342bp、PM的457bp和HPS的821bp的特异性片段。该复合PCR能同时检测到100个每种细菌或50pg的APP或HPS的DNA和500pg的PM的DNA。该三重复合PCR的敏感性同已报道的单PCR一致。该方法的建立对临床上进行这3种疾病的鉴别诊断和混合感染的检测都具有重要意义。 相似文献
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新疆地区4种猪病多重PCR检测方法的建立与初步应用 总被引：1，自引：0，他引：1
本试验旨在建立一种可同时检测猪繁殖与呼吸综合征病毒(porcine reproductive and respiratory syndrome virus,PRRSV)、猪圆环病毒2型(porcine circovirus type 2,PCV2)、猪伪狂犬病病毒(porcine pseudorabies virus,PRV)、猪肺炎支原体(Mycoplasma hyopneumoniae,Mh)的多重PCR检测方法。参考相关文献合成PRRSV、PCV2、PRV、Mh的特异性引物,通过对反应条件优化、特异性、敏感性测定,建立了可同时检测以上4种猪多发传染病的PCR诊断方法。扩增的片段长度分别为424 bp (PRRSV)、490 bp (PCV2)、298 bp (PRV)和360 bp (Mh),对其他常见猪病病原无特异性扩增,采用Multiplex PCR Master Mix对PRRSV、PCV2、PRV、Mh的核酸最低检出量为3个拷贝。采用多重PCR方法对60份临床样本反复检测,与单重PCR相比,4种病原符合率均为100%。结果表明,建立的多重PCR诊断方法可用于猪群中上述4种病原的单一或混合感染的快速鉴别诊断和流行病学调查。 相似文献
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猪传染性胸膜肺炎放线杆菌PCR检测方法的建立及应用 总被引：1，自引：0，他引：1
猪传染性胸膜肺炎是由胸膜肺炎放线杆菌引起猪的一种高度接触性呼吸道传染病,该病可给养猪业造成巨大的经济损失。为有效控制和确诊该病,根据报道的猪胸膜肺炎放线杆菌APXIV毒株的基因序列,合成了2对可扩增长度分别为442 bp和378 bp的特异引物,建立检测胸膜肺炎放线杆菌的巢式PCR方法。利用合成的引物在扩增猪肺疫巴氏杆菌、猪链球菌、副猪嗜血杆菌和大肠杆菌等细菌DNA时,结果均为阴性。用引物检测猪胸膜肺炎放线杆菌的标准菌株可扩增出442 bp和378 bp的特异性条带。表明运用PCR法检测猪胸膜肺炎放线杆菌的特异性和灵敏性均较高,可作为猪传染性胸膜肺炎的快速诊断和流行病学调查的手段。 相似文献
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猪胸膜肺炎放线杆菌、多杀性巴氏杆菌和副猪嗜血杆菌复合PCR检测方法的建立 总被引：2，自引：0，他引：2
目的建立可以同时检测猪胸膜肺炎放线杆菌、多杀性巴氏杆菌和副猪嗜血杆菌的快速而可靠的PCR检测方法。方法和结果根据胸膜肺炎放线杆菌的Apx-VIA基因序列、多杀性巴氏杆菌和副猪嗜血杆菌的16SrRNA基因序列设计5条引物。猪胸膜肺炎放线杆菌、多杀性巴氏杆菌和副猪嗜血杆菌模板的PCR扩增产物大小分别为342bp,485bp和1258bp。复合PCR对1～12型猪胸膜肺炎放线杆菌标准株,6株多杀性巴氏杆菌标准株,1～15型副猪嗜血杆菌以及25株经生化鉴定确认为上述三种细菌的分离株的基因组DNA作为模板进行检测,均获得预期大小的扩增产物。以猪放线杆菌、吲哚放线杆菌等14种常见细菌作为阴性对照进行PCR检测,结果仅有支气管败血波氏杆菌产生了可以和上述三个特异性条带明显区分的PCR产物。复合PCR针对胸膜肺炎放线杆菌、多杀性巴氏杆菌和副猪嗜血杆菌的敏感性分别为14pg、34pg和37pg。结论本研究建立的复合PCR特异性好,敏感性高,可以用于猪胸膜肺炎放线杆菌、多杀性巴氏杆菌和副猪嗜血杆菌的快速检测。 相似文献
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猪胸膜肺炎放线杆菌、肺炎支原体和多杀性巴氏杆菌联合检测芯片的研制及应用 总被引：3，自引：1，他引：2
本研究旨在建立联合检测胸膜肺炎放线杆菌、猪肺炎支原体和多杀性巴氏杆菌的DNA芯片.用7个从3种病菌基因组中扩增出的不同特异性靶DNA制作基因芯片,并对芯片的靶DNA和探针浓度、杂交温度、重复性、特异性和灵敏度进行了研究.结果表明,检测芯片的特异性强,能与测试的李氏放线杆菌、猪鼻支原体和副猪嗜血杆菌等9种病原区分;灵敏度高,在50μL标记反应体系中,能检测到10～50 pg基组DNA,芯片可重复利用.用芯片对44株目标菌的不同型标准菌株、分离株和疫苗株进行了检测.其信号值≥1 000,信号噪音比(SNR)≥6.用芯片对45头病猪和97头健康猪的临床样品选择培养物进行了检测,其检出率分别为多杀性巴氏杆菌71.1%和49.5%、胸膜肺炎放线杆菌42.2%和26.8%、猪肺炎支原体20%和22.7%,混合感染率分别为42.2%和24.7%.在检测临床样品时,芯片法与PCR的符合率为97.8%～100%,与分离鉴定法的符合率为87.6%～95.6%.研究表明,研制的芯片特异性强、敏感性高、可重复使用,是一种能有效用于胸膜肺炎放线杆菌、猪多杀性巴氏杆菌和猪肺炎支原体鉴定和联合检测的新工具. 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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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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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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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