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1.
为建立定量检测猪Janus激酶(JAK)和信号转导和转录激活因子(STAT) mRNA的TaqMan荧光定量PCR检测方法,本研究针对猪JAK-1,JAK-2,STAT-1、STAT-2及管家基因β-actin的序列设计了特异性的引物和探针,分别构建了各自的阳性重组质粒,建立了TaqMan荧光定量PCR方法.结果表明:所建立的检测方法荧光定量PCR扩增均无非特异性产物的产生;检测下限均达1.0×101 copies/μL;批内及批间变异系数均小于2％.利用该方法对猪细小病毒(PPV)感染ST细胞48 h后JAK-1、JAK-2、STAT-1和STAT-2 mRNA的转录水平进行了检测,结果表明:JAK-1和JAK-2转录水平显著上调,STAT-1和STAT-2转录水平呈现下调趋势,本研究建立的TaqMan荧光定量PCR方法特异性强、敏感性高、重复性好,适于猪细胞传导因子JAK和STAT的定量检测. 相似文献
2.
《畜牧与兽医》2020,(8)
为建立快速、灵敏且特异的检测猪圆环病毒3型(porcine circovirus, PCV)的实时荧光定量PCR方法,本研究根据ORF2基因的保守序列设计一对特异性引物和TaqMan探针,通过构建标准品质粒来制作荧光定量PCR标准曲线,优化反应条件,验证敏感性、特异性及重复性,建立了检测PCV3的实时荧光定量PCR方法。结果显示,该方法可以特异性检测出PCV3,而对猪圆环病毒2型、猪瘟病毒、猪繁殖与呼吸综合征病毒、猪流行性腹泻病毒、猪细小病毒、伪狂犬病毒等检测结果均为阴性,表明其具有良好的特异性。用构建的标准品质粒测得的灵敏度可以达到10 copies/μL,重复试验批内及批间变异系数均小于2%,表明其具有良好的灵敏性和重复性。以上结果表明,本研究建立的TaqMan荧光定量PCR检测方法具有快速、灵敏性高、特异性强的优点,可为我国PCV3型的早期检测及相关研究工作提供可靠的技术支持。 相似文献
3.
应用TaqMan荧光定量PCR检测H3N8亚型马流感病毒 总被引：2，自引：1，他引：1
为建立马流感病毒(ETV)的检测方法,本试验针对H3N8亚型ETV血凝素(HA)基因高度保守序列设计并合成了2对引物和1条TaqMan荧光探针,建立了TaqMan荧光定量PCR方法.经用TaqMan荧光定量PCR、RT-PCR和病毒分离方法分别检测新疆等省135份疑似EIV马鼻拭子样品,其结果表明:3种方法的马流感检出率分别为54.07%、37.78%、0.89%;TaqMan荧光定量PCR可检出马流感病毒基因组RNA的灵敏度可达10拷贝/反应.而且与其他马呼吸道病毒均无交叉反应,具有良好的特异性、敏感性和重复性.该方法为H3N8亚型马流感的早期诊断及分子流行病学调查等提供了一种新的快速、准确的定量检测技术. 相似文献
4.
《中国兽医杂志》2019,(5)
为建立一种准确、特异、高效、快速的非洲猪瘟病毒定量检测方法,本研究根据非洲猪瘟病毒(African swine fever virus, ASFV)早期表达基因K196R的基因序列,设计了TaqMan荧光定量PCR引物及探针,通过优化退火温度、引物及探针浓度,建立了快速检测ASFV的TaqMan荧光定量PCR检测方法。结果表明,该方法选择的引物具有高度灵敏性和特异性,以构建的重组质粒为标准品建立的TaqMan荧光定量PCR方法的标准曲线具有良好的线性关系(R~2=0.998),对ASFV核酸最低检测下限为1.3拷贝,且与猪伪狂犬病病毒、猪细小病毒、猪圆环病毒2型等多种病原不存在交叉反应。本研究建立的K196R基因实时荧光定量PCR检测方法为非洲猪瘟疫情提供了一种新型、灵敏和特异的早期检测方法。 相似文献
5.
《中国预防兽医学报》2017,(2)
为建立同时快速鉴别检测牛病毒性腹泻病毒(BVDV)、牛传染性支气管炎病毒(IBRV)和口蹄疫病毒(FMDV)的方法,本研究根据高度保守的BVDV的5'UTR基因、IBRV的g B基因和FMDV的3D基因,分别设计了3对对应的特异性引物和3种不同发光基团标记的Taq Man探针,建立了同时检测这3种病毒的多重荧光定量PCR的方法。并对其反应条件进行优化。结果表明,该多重荧光定量PCR方法能够特异性检测出BVDV、IBRV和FMDV,而对BTV等检测结果均为阴性,对BVDV、IBRV、FMDV的最低检测量分别为194拷贝/μL、208拷贝/μL和150拷贝/μL。而且组内和组间变异系数均低于0.85%。本研究所建立的多重荧光定量PCR具有方便、快速、特异性好、灵敏度高、重复性好等优点,能够用于BVDV、IBRV和FMDV的同时检测。 相似文献
6.
为提高鸭坦布苏病毒(DTMUV)TaqMan荧光定量PCR方法的敏感性和简化反应条件,本研究在前期建立的TaqMan荧光定量PCR方法的基础上,设计一条特异性的反转录引物,优化反应体系,建立了简便、快速、敏感的TaqMan荧光定量PCR方法。该荧光定量PCR方法最低检测限为10拷贝,敏感性是未优化的荧光定量PCR方法的5倍、是普通PCR方法的100倍,并且该方法对DTMUV的最低检测限是0.01半数鸡胚致死量(ELD50)。通过批内和批间实验的变异系数表明优化的荧光定量PCR方法的重复性比未优化的荧光定量PCR方法好。通过该优化的荧光定量PCR方法检测其它常见的鸭病病毒的DNAs或RNAs,证明了该方法特异性好。对现地60份疑似DTMUV的样品进行检测,用该方法检出57份样品为阳性,明显高于普通PCR,并且在区分35Ct值左右样品试验中敏感性优于未优化的荧光定量PCR方法。该优化的DTMUV TaqMan荧光定量PCR方法更适用于DTMUV的快速定量流行病学诊断。 相似文献
7.
为建立检测PHoV的TaqMan荧光定量PCR方法,本研究根据PHoV的VP2基因序列设计引物和探针,以梯度稀释的含有VP2基因的重组质粒作为标准品,进行定量PCR反应.结果显示,该方法的检测灵敏度为10拷贝;而且该检测方法特异性较好,与猪的其他病毒核酸均无交叉反应;批内和批间的变异系数低于3.29％,表明该方法的重复性较好.对华东地区采集的225份临床样品进行检测,结果显示,PHoV的阳性率为14.2％.本研究建立的荧光定量PCR方法灵敏度高、特异性好,可以为PHoV的流行病学调查和发病机制等研究提供可靠的工具. 相似文献
8.
《中国预防兽医学报》2018,(11)
为建立同时快速鉴别检测羊痘病毒属病毒(CaPV)的方法,本研究设计了一对针对CaPV的通用引物和3条特异性的TaqMan MGB探针,并对其反应条件进行优化,建立了单管同时鉴别检测3种病毒的多重TaqMan MGB荧光定量PCR方法。结果显示,该多重荧光定量PCR方法仅对牛结节性皮肤病病毒(LSDV)、山羊痘病毒(GTPV)和绵羊痘病毒(SPPV)有特异性扩增,而对牛痘病毒等其它相关病毒核酸无扩增。在10~2拷贝/μL～107拷贝/μL浓度范围内有良好的线性关系;对LSDV、 GTPV、SPPV的最低检测限分别为14.8拷贝/μL、32.9拷贝/μL、26.7拷贝/μL。标准曲线相关系数(R2)均为0.999。批内及批间变异系数均小于1.5%。应用本研究建立的方法和OIE陆生动物手册推荐普通PCR方法分别对185份临床样品和67份模拟样品进行检测,该方法检出率比普通PCR方法高约1.6%。本研究建立的CaPV多重TaqMan-MGB荧光定量PCR方法具有特异性强、敏感性高、重复性好、操作简便快速等优点,适用于CaPV临床样品的快速鉴别检测。 相似文献
9.
10.
为了建立快速检测水貂犬瘟热病毒的TaqMan荧光定量PCR方法,试验根据犬瘟热病毒核衣壳蛋白基因的保守区域设计一对特异引物和探针,优化体系反应条件,并进行敏感性试验、特异性试验、重复性试验和水貂外周血淋巴细胞感染犬瘟热病毒后的检测。结果表明:建立的TaqMan荧光定量PCR检测方法最低检出量为3.96个拷贝;与水貂细小病毒、水貂阿留申病毒均无交叉反应;批内重复和批间重复循环阈值(Ct)变异系数均小于1%;应用该方法可成功检出水貂外周血淋巴细胞中的犬瘟热病毒。说明建立的检测方法敏感性、特异性、重复性好,可以用于水貂犬瘟热病毒的临床检测。 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.
Rassnick KM Gieger TL Williams LE Ruslander DM Northrup NC Kristal O Myers NC Moore AS 《Journal of veterinary internal medicine / American College of Veterinary Internal Medicine》2001,15(3):196-199
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.
H Hayashi T Yoshimura J Y Chen 《Comparative immunology, microbiology and infectious diseases》1985,8(2):73-87
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.
Davis MS Willard MD Nelson SL Mandsager RE McKiernan BS Mansell JK Lehenbauer TW 《Journal of veterinary internal medicine / American College of Veterinary Internal Medicine》2003,17(3):311-314
Human and equine athletes are reported to have a high prevalence of gastric disease, and anecdotal evidence suggests a similar phenomenon applies to racing sled dogs. To investigate the prevalence of gastric disease in racing sled dogs, we conducted 2 gastroscopy studies on dogs competing in the annual Iditarod Sled Dog Race. A pilot study of dogs that were either dropped from the 2000 Iditarod Sled Dog Race because of illness or that finished the race indicated that, approximately 5 days after competing, 10 of 28 dogs (35%) had endoscopic evidence of gastric ulceration, erosion, or hemorrhage. The next year, an endoscopic study of 73 dogs participating in the 2001 Iditarod race was performed in order to evaluate a larger population of dogs. Data from 70 of these dogs could be used; 34 (48.5%) had ulceration, erosion, gastric hemorrhage, or some combination of these findings. When this group of 70 dogs was compared retrospectively to a control group of 87 dogs presented to the Texas A&M University (TAMU) Veterinary Medical Teaching Hospital, the Iditarod sled dogs had a significantly higher prevalence (P = .049) of gastric lesions. These findings suggest that, similar to athletes of other species, elite canine athletes have an increased prevalence of gastric disease compared to the canine population at large. 相似文献
17.
Furosemide continuous rate infusion in the horse: evaluation of enhanced efficacy and reduced side effects 总被引：1，自引：0，他引：1
Johansson AM Gardner SY Levine JF Papich MG LaFevers DH Fuquay LR Reagan VH Atkins CE 《Journal of veterinary internal medicine / American College of Veterinary Internal Medicine》2003,17(6):887-895
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
18.
B. Cinar Kul N. Bilgen J.A. Lenstra O. Korkmaz Agaoglu B. Akyuz O. Ertugrul 《Zeitschrift für Tierzüchtung und Züchtungsbiologie》2015,132(6):449-453
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
19.
Ling GV Thurmond MC Choi YK Franti CE Ruby AL Johnson DL 《Journal of veterinary internal medicine / American College of Veterinary Internal Medicine》2003,17(6):817-823
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
20.
<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