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1.
对伪狂犬病病毒（PRV）、猪细小病毒（PPV）和流行性乙型脑炎病毒（JEV）检测基因芯片的制备及该芯片的检测技术进行了研究。选定靶基因最佳点样质量浓度为200 mg/L,用基因芯片点样仪将其点制在氨基化基片上,经干燥、水合、紫外线交联和洗涤后,成功制备了PRV-PPV-JEV检测基因芯片。以CY3荧光素标记的dCTP经PCR扩增制备探针,对芯片的质量进行了评价。结果表明,制备的芯片质量好,探针最佳使用质量浓度为3 000μg/L,芯片系统检测灵敏度可达3μg/L。该芯片可同时检测PRV、PPV和JEV,其灵敏度高、特异性强,芯片可重复使用,室温下至少可保存4个月。  相似文献   

2.
根据GenBank中已发表的猪圆环病毒2型(PCV2)、猪伪狂犬病毒(PRV)、猪细小病毒(PPV)有关基因序列,通过序列比对及参考相关文献,设计了3对特异性引物。通过优化,建立了可同时检测PCV2、PRV及PPV 3种病毒的多重PCR方法。结果表明,该方法能同时检测到PCV2、PRV及PPV的最低核酸量分别为40pg、72pg、46pg。对猪圆环病毒3型(PCV3)、猪细环病毒(TTSuV)、猪流行腹泻病毒(PEDV)、猪蓝耳病毒(PRRSV)、猪瘟病毒(CSFV)无扩增。应用该方法对87份临床样品进行检测,PCV2阳性检出率为28.7%(25/87),PRV野毒检出率为11.5%(10/87),PPV检出率为12.6%(11/87)。其中2种以上病毒混合感染阳性率为13.8%(12/87),3种病毒同时感染的阳性率为2.3%(2/87),其结果也与单一PCR结果一致。研究结果提示,该方法可用于PCV2、PRV、PPV三种病原的单一感染或混合感染的鉴别诊断。  相似文献   

3.
猪传染性胃肠炎病毒检测基因芯片的构建   总被引：6，自引：0，他引：6  
采用PCR扩增制备TGEV的靶基因并进行纯化,对基因芯片的最佳靶基因点样质量浓度、探针质量浓度、杂交温度、杂交时间进行筛选,选择构建检测芯片的最适靶基因,进行基因芯片探针最佳标记方法试验.结果表明,质粒PCR扩增和采用异丙醇沉淀纯化的靶基因质量好,基因芯片最佳靶基因点样质量浓度为200mg/L,最佳探针质量浓度为3 000 μg/L,预杂交时间为1 h,杂交时间为3～6 h,杂交温度为48℃,以S、S3、sM、M、N、ORF7、POL等7个靶基因为构建TGEV检测芯片的最适靶基因,确定了多重PCR扩增标记TGEV探钟的最佳体系,Cy3-dCTP标记浓度为2.5 μmol/L,构建的TGEV检测芯片与标记的混合探针杂交效果好.  相似文献   

4.
试验旨在了解广西猪主要病毒性繁殖障碍疫病的发生态势,为预防和控制此类疫病提供科学依据。采用RT-PCR/PCR方法对2014年3月至2018年2月采自广西的349份流产胎儿样品进行猪瘟病毒(CSFV)、猪繁殖与呼吸综合征病毒(PRRSV)、猪圆环病毒2型(PCV2)、猪伪狂犬病病毒(PRV)、猪细小病毒(PPV)及猪乙型脑炎病毒(JEV)6种主要病毒性繁殖障碍疫病病原的检测并分析其阳性检出率在不同年份、季节之间的差异及病原混合感染情况。结果显示,2014-2017年,6种病原的猪场阳性率从高到低依次为JEV(36.00%)、PRRSV(16.00%)、PCV2(9.00%)、PRV(7.00%)、PPV(6.00%)和CFSV(5.00%),样品阳性检出率从高到低依次为JEV(27.06%)、PRRSV(6.18%)、PRV(5.88%)、PCV2(3.82%)、CSFV(2.35%)和PPV(2.35%)。季节因素对6种病毒性繁殖障碍疫病的发生有一定影响:JEV在夏季、秋季检出率较高,春季、冬季检出率较低;PRRSV一年四季检出率均较高,且无明显差异;PCV2在春季检出率较高,其次是夏季和冬季,秋季检出率较低;PRV在春季、冬季检出率较高,夏季、秋季检出率均为0;PPV在夏季检出率较高,其次是春季和秋季,冬季检出率为0;CSFV在冬季检出率最高,其次是夏季,春季和秋季检出率均为0。流产胎儿以单一感染为主,6种病毒均存在不同程度单一感染情况,单一感染率JEV最高(23.82%)、PCV2最低(0.59%);混合感染以双重感染为主,其中以2016年双重感染情况较多,且感染组合多样;三重感染情况较少、仅在2016年送检样品中发现1例,未发现四重及四重以上感染情况。调查结果表明,2014-2017年,JEV均有较高的阳性检出率,已成为广西部分猪场母猪流产最主要病毒性病原。PRRSV和PRV仍是目前引起广西猪场母猪流产的主要病毒性病原。  相似文献   

5.
为了检测猪伪狂犬病病毒和猪博卡病毒混合感染的情况,研究建立了快速,简便,灵敏度高,特异性强的鉴别猪伪狂犬病病毒(PRV)和猪博卡病毒(PBoV)的双重检测方法。针对PRV gE基因和PBoV NS1基因的序列,分别设计了2对特异性引物,通过对引物浓度、模板加入量、退火温度等的优化,建立了快速鉴别PRV和PBoV的双重PCR检测方法,并进行了特异性和敏感性试验。特异性试验表明,该方法可以扩增出PRV gE长度为316 bp和PBoV NS1长度为996 bp的目的片段,对大肠杆菌、猪圆环病毒Ⅱ型、猪捷申病毒、猪瘟病毒、猪繁殖与呼吸综合征病毒均无扩增;敏感性试验表明,PRV、PBoV的最低核酸检出量分别为6.44×10~2拷贝和9.51×10~3拷贝,与单一PCR敏感性相同。应用该方法对49份送检病料样品进行检测,其中PRV和PBoV的单纯感染检出率分别为34.69%和14.28%,混合感染检出率为8.16%,与单一PCR检测结果符合率为93.88%。  相似文献   

6.
用地高辛标记核酸探针检测猪细小病毒的研究   总被引：3，自引：0，他引：3  
根据猪细小病毒(PPV)基因组序列,设计引物克隆了PPV YK株保守序列NS1基因,将NS1基因纯化,利用地高辛标记制备了NS1基因PCR产物探针和克隆NS1探针,两种核酸探针的标记都达到了0.1 pg/μL。特异性试验结果表明,这两种探针都能与PPV DNA发生特异的阳性杂交,而与对照的PRV、PCV、PRRSV和HCV的核酸杂交呈阴性。敏感性试验结果表明,克隆NS1探针敏感度为0.5 pg/μL,NS1基因PCR产物探针敏感度为1 pg/μL。综上所述,这两种DIG标记探针特异性强,敏感性高,可用于PPV的检测。  相似文献   

7.
广西猪细小病毒与PRRSV、CSFV、PCV2、PRV混合感染的检测   总被引：7，自引：0，他引：7  
应用PCR技术,对广西南宁、玉林、贵港、柳州、钦州和桂林6个市的10个规模化猪场送检的141份病猪组织样品进行猪细小病毒(PPV)的检测;同时,对鉴定为PPV阳性的样品进行了猪繁殖与呼吸综合征病毒(PRRSV)、猪瘟病毒(CS-FV)、猪圆环病毒2型(PCV2)和猪伪狂犬病毒(PRV)的检测,以确定猪群中PPV与其它病毒混和感染情况。结果,所有样品诸病毒的阳性检出率达17.73%(25/141);流产胎儿PPV阳性率为19.57%(9/46),繁殖障碍母猪扁桃体的PPV阳性率为17.74%(11/62),发病断奶仔猪组织病料的PPV阳性率为15.15%(5/33),PPV与PRRSV、CSFV、PCV2和/或PRV4种病毒均有混合感染现象,混合感染样品占PPV阳性样品的48.00%(12/25)。  相似文献   

8.
根据GenBank中PRRSV ORF6、PCV2 ORF2、PPV NS1、PRV gB、SIV M、JEV NS1、CSFV E2等基因的部分片段,分别设计特异性引物,用实验室已保存的病毒及阳性病料,通过对7个病毒单一PCR的退火温度和敏感性进行优化和检测,再选择合适的多重PCR缓冲体系及酶,优化退火温度、模板及引物浓度,最后检测多重PCR的敏感性、特异性及重复性,建立一种能够同时且快速鉴别PRRSV、PCV2、PPV、PRV、SIV、JEV和CSFV这7种常见猪呼吸道和繁殖障碍类病毒的多重PCR检测方法,并用其对2017-2019年贵州部分地区的150份临床病料进行检测。多重PCR特异性检测结果表明:从混合病毒基因组中分别扩增出大小为224,353,424,549,684,831,1 137 bp的特异性片段,未扩增出E.coli、APP、PEDV及正常组织的DNA/RNA;敏感性试验表明,该方法对病毒的最低检测量分别为PRRSV 94 pg、PCV2 66 pg、PPV 68 pg、PRV 20 pg、SIV 35 pg、JEV 17 pg、CSFV 14 pg;重复性试验表明:相同条件下的6次重复试验均能扩增出7个病毒的目的条带;应用试验表明:150份临床病料中二重感染阳性率高达66.00%(99/150);三重感染阳性率达11.33%(17/150);四重感染阳性率达2.00%(3/150);五重感染阳性率达0.67%(1/150);六重感染阳性率为0,七重感染阳性率为0,阳性PRRSV、PCV2、PPV、JEV、SIV的多重PCR与单一PCR的符合率均为100.00%,阳性CSFV和PRV的符合率为90.00%以上,研究建立的多重PCR检测方法具有敏感性高、特异性强、重复性好的特点,对猪场猪呼吸和繁殖障碍类病毒病混合感染的快速诊断具有十分重要的意义。  相似文献   

9.
根据Genbank上已发表的猪伪狂犬病毒(PRV)、猪细小病毒(PPV)和猪流行性乙型脑炎病毒(JEV)基因序列，用Array Designer2．0软件设计并合成了PRV gB、PPV VP2、JEV C基因片段的3对引物，以PRV Fa株、PPV B2株、JEV SA14-14-2株细胞培养物提取棱酸人为混合后作为模板，筛选PCR反应条件，扩增出长度分别为324bp、471bp和238bp的3条特异性片段，建立了检测3种病毒的多重PCR方法。应用该方法对PRVFa、PRV渝A、8F20、BUK株、德国Bartha-K61株、PRV Ea株、PRV疫苗毒，PPV132株、PPV SR标准株SR-1、SR-2、SR-3，JEV SA14-14-2株的病毒核酸进行扩增，均获得了特异性的目的片段，而扩增PRRSV、CFSV、PCV-2及相应的培养细胞核酸结果均为阴性。对PRV Fa、PPV B2和JEV SA14-14-2的最低检出量分别为17．5pg、13．7pg和20pg的DNA或cDNA。该试验方法适合对PRV、PPV和JEV的联合检测和鉴别诊断。  相似文献   

10.
本研究旨在建立一种能快速、灵敏、同时检测出猪细小病毒与猪伪狂犬病毒的基于SYBR GreenⅠ实时荧光定量PCR方法。参照GenBank中登录的相关基因序列,设计了2对引物分别用于扩增PRV gH基因与PPV NS1基因的部分片段。将测序正确的PRV gH基因与PPV NS1基因片段克隆入pGEM-T Easy载体,转化大肠杆菌DH5α,经测序鉴定后得到阳性重组质粒,作为标准品模板建立SYBR GreenⅠ荧光定量PCR标准曲线和熔解曲线,并对其灵敏性、特异性和重复性进行验证。结果表明,猪细小病毒与猪伪狂犬病毒荧光定量PCR的标准曲线的Tm值分别为80.9℃和86.5℃,熔解曲线特异,灵敏度分别可达248拷贝/μL和160拷贝/μL,是普通PCR检测方法的100倍。本次建立的猪细小病毒与猪伪狂犬病毒荧光定量PCR检测方法实现了2种病毒的同时检测,能够对PRV、PPV混合感染的临床病料进行快速诊断。  相似文献   

11.
试验选用96头平均体重14.82 kg左右的杜×长×大断奶仔猪,随机分成4组,每组3栏,每栏8头(公母各半)。对照组饲喂基础日粮,试验1、2、3组分别添加1%80目白术、0.2%白术多糖和1%微米白术。试验期30 d。结果表明:在生长性能方面,与对照组相比,1%微米白术添加组可显著提高日增重(P0.05)、降低饲料增重比和腹泻率,而且效果优于1%80目白术组和0.2%白术多糖组,在肠道形态和肠道微生态区系方面,与对照组相比,日粮添加1%80目白术、0.2%白术多糖、1%微米白术均可不同程度的提高十二指肠和空肠的绒毛高度,加深十二指肠和空肠的隐窝深度,并且增加肠道微生态区系的多样性,其中以1%微米白术添加组的效果最佳。  相似文献   

12.
Sissay, M.M., Uggla, A. and Waller, P.J., XXXX. Prevalence and seasonal incidence of nematode parasites and fluke infections of sheep and goats in eastern Ethiopia. Tropical Animal Health and Production, XXXX. A 2-year abattoir survey was carried out to determine the prevalence, abundance and seasonal incidence of gastro-intestinal (GI) nematodes and trematodes (flukes) of sheep and goats in the semi-arid zone of eastern Ethiopia. During May 2003 to April 2005, viscera including liver, lungs and GI tracts were collected from 655 sheep and 632 goats slaughtered at 4 abattoirs located in the towns of Haramaya, Harar, Dire Dawa and Jijiga in eastern Ethiopia. All animals were raised in the farming areas located within the community boundaries for each town. Collected materials were transported within 24 h to the parasitology laboratory of Haramaya University for immediate processing. Thirteen species belonging to 9 genera of GI nematodes (Haemonchus contortus, Trichostrongylus axei, T. colubriformis, T. vitrinus, Nematodirus filicollis, N. spathiger, Oesophagostomum columbianum, O. venulosum, Strongyloides papillosus, Bunostomum trigonocephalum, Trichuris ovis, Cooperia curticei and Chabertia ovina), and 4 species belonging to 3 genera of trematodes (Fasciola hepatica, F. gigantica, Paramphistomum {Calicohoron} microbothrium and Dicrocoelium dendriticum) were recorded in both sheep and goats. All animals in this investigation were infected with multiple species to varying degrees. The mean burdens of adult nematodes were generally moderate in both sheep and goats and showed patterns of seasonal abundance that corresponded with the bi-modal annual rainfall pattern, with highest burdens around the middle of the rainy season. In both sheep and goats there were significant differences in the mean worm burdens and abundance of the different nematode species between the four geographic locations, with worm burdens in the Haramaya and Harar areas greater than those observed in the Dire Dawa and Jijiga locations. Similar seasonal variations were also observed in the prevalence of flukes. But there were no significant differences in the prevalence of each fluke species between the four locations. Overall, the results showed that Haemonchus, Trichostrongylus, Nematodirus, Oesophagostomum, Fasciola and Paramphistomum species were the most abundant helminth parasites of sheep and goats in eastern Ethiopia.  相似文献   

13.
Tropical Animal Health and Production -  相似文献   

14.
1-(2-Chloroethyl)3-cyclohexyl-1-nitrosourea (CCNU) is an alkylating agent in the nitrosourea subclass. A prospective evaluation of CCNU was done to determine the maximally tolerated dosage of CCNU in tumor-bearing cats. Response data were obtained when available. Twenty-five cats were treated with CCNU at a dosage of 50-60 mg/m3 body surface area. Complete hematologic data were available for 13 cats. Neutropenia was the acute dose-limiting toxicity. The median neutrophil count at the nadir was 1,000 cells/microL (mean, 2,433 cells/microL; range, 0-9,694 cells/microL). The time of neutrophil nadir was variable, occurring 7-28 days after treatment, and counts sometimes did not return to normal for up to 14 days after the nadir. Based on these findings, a 6-week dosing interval and weekly hematologic monitoring after the 1st treatment with CCNU are recommended. The nadir of the platelet count may occur 14-21 days after treatment. The median platelet count at the nadir was 43,500 cells/microL. No gastrointestinal, renal, or hepatic toxicities were observed after a single CCNU treatment, and additional studies to evaluate the potential for cumulative toxicity should be performed. Five cats with lymphoma and 1 cat with mast cell tumor had measurable responses to CCNU. Phase II studies to evaluate antitumor activity should be completed with a dosing regimen of 50-60 mg/m3 every 6 weeks.  相似文献   

15.
Our particular attention in this article was given to natural mediators for macrophages isolated from the sites of tissue injury. A number of chemotactic factors, which may satisfy many criteria making them acceptable as inflammatory leucocyte chemotactic factors, has been separated. Among them, our laboratory has isolated three macrophage (monocyte) chemotactic factors (MCF-a, -b and -c). Their purification, characterization and functional specificity are discussed.  相似文献   

16.
Continuous rate infusion (CRI) of furosemide in humans is considered superior to intermittent administration (IA). This study examined whether furosemide CRI, compared with IA, would increase diuretic efficacy with decreased fluid and electrolyte fluctuations and activation of the renin-angiotensin-aldosterone system (RAAS) in the horse. Five mares were used in a crossover-design study. During a 24-hour period, each horse received a total of 3 mg/kg furosemide by either CRI (0.12 mg/kg/h preceded by a loading dose of 0.12 mg/kg IV) or IA (1 mg/kg IV q8h). There was not a statistically significant difference in urine volume over 24 hours between methods; however, urine volume was significantly greater after CRI compared with IA during the first 8 hours ([median 25th percentile, 75th percentile]: 9.6 L [8.9, 14.4] for CRI versus 5.9 L [5.3, 6.0] for IA). CRI produced a more uniform urine flow, decreased fluctuations in plasma volume, and suppressed renal concentrating ability throughout the infusion period. Potassium, Ca, and Cl excretion was greater during CRI than IA (1,133 mmol [1.110, 1,229] versus 764 mmol [709, 904], 102.7 mmol [96.0, 117.2] versus 73.3 mmol [65.0, 73.5], and 1,776 mmol [1,657, 2.378] versus 1,596 mmol [1,457, 1,767], respectively). Elimination half-lives of furosemide were 1.35 and 0.47 hours for CRI and IA, respectively. The area under the excretion rate curve was 1,285.7 and 184.2 mL x mg/mL for CRI and IA, respectively. Furosemide CRI (0.12 mg/kg/h) for 8 hours, preceded by a loading dose (0.12 mg/kg), is recommended when profound diuresis is needed acutely in horses.  相似文献   

17.
Genetic variations in chromosome Y are enabling researchers to identify paternal lineages, which are informative for introgressions and migrations. In this study, the male‐specific region markers, sex‐determining region‐Y (SRY), amelogenin (AMELY) and zinc finger (ZFY) were analysed in seven Turkish native goat breeds, Angora, Kilis, Hair, Honaml?, Norduz, Gürcü and Abaza. A SNP in the ZFY gene defined a new haplotype Y2C. All domestic haplogroups originate from Capra aegagrus, while the finding of Y1A, Y1B, Y2A and Y2C in 32, 4, 126 and 2 Turkish domestic goats, respectively, appears to indicate a predomestic origin of the major haplotypes. The occurrence of four haplotypes in the Hair goat and, in contrast, a frequency of 96% of Y1A in the Kilis breed illustrate that Y‐chromosomal variants have a more breed‐dependent distribution than mitochondrial or autosomal DNA. This probably reflects male founder effects, but a role in adaptation cannot be excluded.  相似文献   

18.
The objective of this report was to characterize 20-year changes in proportion of calcium oxalate (CaOx) calculi and struvite calculi in dogs, and associations with breed, age, and sex. In this retrospective study, results of analysis of urinary calculi from dogs were reviewed for specimens received between July 1, 1981, and December 31, 2001. Breed, sex, age, year of submission of the specimen, and mineral type(s) were analyzed statistically. CaOx or Struvite or both were contained in 18,966 of 20,884 (91%) specimens. For both sexes, a 20-year statistically significant increase was observed in the proportion of calculus specimens that contained CaOx. The increase in this proportion was greater in females (1% to 31%) than in males (18% to 82%). From 1998 to 2001, when proportions may have plateaued, the odds of specimens containing CaOx were markedly higher in 18 breeds, markedly lower in 5 breeds, and not significantly different in 13 breeds compared with crossbreds. For both sexes, a 20-year statistically significant decrease was observed in the proportion of calculus specimens that contained struvite. This decrease in proportion was greater for males (79-16%) than for females (97-68%). From 1998 to 2001, when proportions plateaued, the odds of calculi containing struvite were markedly lower in 20 breeds, markedly higher in 1 breed, and not significantly different in 15 breeds when compared with crossbreds. Breed, age, and sex were associated statistically with CaOx or struvite urolithiasis. In conclusion, there appears to have been a long-term increase in the proportion of specimens of canine urinary calculi that contain CaOx as well as a long-term decrease in the proportion of specimens of calculi that contain struvite for both male and female dogs. The rate of change appeared to begin leveling off in the period 1998 to 2001. The recent proportion of dogs with either CaOx- or struvite-associated urolithiasis may depend on breed, age, and sex, and on interactions among these 3 factors.  相似文献   

19.
<正>1.INTRODUCTION Ensuring transportation network security is one Of the most daunting challenges confronting homeland security agencies today.Significant research has been dedicated.To model and analyze the vulnerability of transportation systems,while notably fewer studies propose specific strategies for deploying defensive technologies to safeguard these systems.  相似文献   

20.
The epidemiology of H. placei and of other gastrointestinal nematodes in yearling dairy cattle was examined on two farms in Kiambu District, central Kenya during each of 13 one-month periods from April 1993 to April 1994. On each farm, 32 newly weaned dairy calves were given a single dose of albendazole and then placed on experimental pastures. Twelve of the animals were designated for bi-monthly slaughter (n = 2) and analysis of worm population characteristics and 20 were designated for blood and faecal collection and for weighing. Two parasite-free tracer calves were grazed alongside the weaner calves each month throughout the study period and were also slaughtered for analysis of worm populations. Faecal egg counts, haematological and serum pepsinogen determinations, herbage larval counts, and animal live weight changes were recorded monthly. The study revealed that Haemonchus placei, Trichostrongylus axei, Cooperia spp. and Oesophagostomum radiatum were responsible for parasitic gastroenteritis and that H. placei was the predominant nematode present in the young cattle on both farms. Faecal egg counts from resident cattle and necropsy worm counts revealed that pasture larval levels were directly related to the amount of rainfall. The total worm burdens in the animals were highest during the rainy season (March–June and October–December) and lowest during the dry seasons (July–September and January–February). The very low recovery of immature larvae of H. placei from the tracer calves indicated that arrested development is not a feature of the life cycle of this parasite in central Kenya. The maintenance of the parasite population depended on continuous cycling of infection between the host and the pasture. The agroclimatic conditions of the study area were such that, in general, favourable weather conditions for the development and survival of the free-living stages of gastrointestinal nematodes existed all year round.  相似文献   

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