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1.
本文建立了快速、简便、微量、高特异性的检测鸡克雷伯氏杆菌病玻片凝集试验。测得最适凝集原最适凝集原量为50亿个菌/毫升,用来检测鸡克雷伯氏杆菌标准株和分离株的兔、抗血清效价分别为256-512(标准株)、512-1024(分离株)。临169分水岭分鸡粪样,825份鸡血清,阳性率分别为28.40%(48/169)、29.45(243/825),而用细菌分离生化鉴定鸡粪阳性率为29.59(50/169)  相似文献   

2.
进口动物性饲料中沙门氏菌的分离鉴定   总被引:5,自引:0,他引:5  
通过常规分离培养鉴定技术,对上海口岸2001年1-6月份进口的动物性饲料498份进行了沙门氏菌的分离鉴定。结果共分离到沙门氏菌23株,分离率为4.62%(23/498)。其中,鱼粉164份,阳性6份,阳性率3.66%(6/164);肉骨粉86份,阳性12份,阳性率为13.95%(12/86);明虾壳27份,阳性5份,阳性率18.52%(5/27);乳清粉和饲料添加剂类221份,阳性0份,阳性率0%。最终对21株沙门氏菌进行定型。  相似文献   

3.
陕西省鸡胚源性大肠杆菌的分离鉴定   总被引:9,自引:0,他引:9  
从陕西关中六个种鸡场的198只死胚及出壳弱雏中分离出98株大肠杆菌,平均分离率为49.5%。通过小鸡致病力试验,筛选出63株具有致病力的菌株,占分离菌的64.3%。生化鉴定表明,这些致病菌均符合大肠埃希氏菌的生化特性。血清型鉴定可确定6种,其中以O1、O2、O78为主,分别占鉴定菌的44.4%(28/63)、33.3%(21/63)和9.5%(6/63),另三种为O22、O75、O89共占8.0%(5/63)。有3株未定型,占4.8%(3/63)。用16种抗生素进行药敏试验,对先锋霉素V和恩诺沙星高度敏感,对羧苄青霉素、氨苄青霉素、氯霉素、卡那霉素、磺胺嘧啶、新霉素、链霉素中度敏感。  相似文献   

4.
为了解石河子地区部分规模化奶牛场奶牛隐性乳房炎源金黄色葡萄球菌的生物学特性及药物敏感性,为科学防治该病的发生提供参考依据,本试验选择了石河子周边地区不同规模的11个奶牛场,通过采用LMT法检测其隐性乳房炎的感染率,并从采集到的313份隐性乳房炎奶样中分离鉴定金黄色葡萄球菌,同时采用纸片扩散法(K-B法)对分离株的药物敏感性进行了分析。结果表明,试验区奶牛隐性乳房炎的总阳性率为44.56%(869/1950),金黄色葡萄球菌的分离率为44.71%(93/208);药敏试验结果表明,大部分菌株对阿莫西林、红霉素、四环素表现为高度耐药,耐药率分别为92.30%(24/26)、80.77%(21/26)、80.77%(21/26);对庆大霉素、氧氟沙星、头孢唑啉表现为敏感,敏感率分别为57.70%(15/26)、76.92%(20/26)、76.92%(20/26);部分菌株对克林霉素中度敏感,敏感率为46.15%(12/26)。建议在选用抗生素治疗奶牛隐性乳房炎时,以流行菌株的药物敏感性为依据。  相似文献   

5.
本研究探索建立一种新的基因芯片方法检测和鉴定猪肺炎支原体。在猪肺炎支原体的16SrRNA、P36和P463个基因内设计3个靶基因片段,用PCR标记Cy3探针,建立了用于检测和鉴定猪肺炎支原体的检测芯片。试验结果显示,猪肺炎支原体与检测芯片的3个靶基因(Mhp-16S、Mhp-P46和Mhp-P36)杂交呈阳性,猪鼻支原体只与Mhp-16S靶基因杂交呈阳性,其它所测细菌和病毒不与检测芯片的靶基因杂交。检测芯片的检测灵敏度和PCR相同,能达到10pg基因组DNA/50μL标记体系或反应体系。用检测芯片鉴定了3株疑似猪肺炎支原体临床分离株,结果有2株为猪肺炎支原体,1株为其它猪支原体。用检测芯片、P36-PCR和分离鉴定分别对45头病猪临床样品选择混合培养物中的猪肺炎支原体进行了检测,结果它们的检出率分别为20%(9/45)、22.2%(10/45)和8.9%(4/45)。检测芯片还检出有26.7%(12/45)的病猪感染了其它猪支原体。试验结果表明,所建立的检测芯片是一种快速检测和鉴定猪肺炎支原体的敏感特异性方法。  相似文献   

6.
动物性饲料中沙门氏菌的分离鉴定   总被引:12,自引:0,他引:12  
通过常规分离培养鉴定技术,对饲料厂成品饲料102份,鱼粉97份、肉骨粉98份进行了沙门氏菌的分离鉴定,结果从297份样品中到沙门氏菌45株分离率15.2%(45/297)。成品饲料阳性18份,阳性率17.6%(13/102)。鱼粉阳性14份,阳性率为14.4%(14/97),肉骨粉134份,阳性率为13.3%(13/98)。  相似文献   

7.
陕西省猪伪狂犬病血清学调查   总被引:4,自引:0,他引:4  
2000年1-月份,在陕西省11个地区(市)的59个县(市)对7458份种猪血清进行了猪伪狂犬病抗体乳胶凝集试验。共检出抗体阳性血清1394份,阳性率为18.69%,对其中44个县(市)的5089份血清进行了详细统计,母猪的血清抗体阳性率为14.85%(55/3662),公猪的血清抗体阳性率为18.81%(120/638),后血母猪的血清抗体阳性率为20.43%(141/690),后备公猪的血清抗体阳性率为28.28%(28/99)。  相似文献   

8.
动物园珍稀野生动物弓形虫感染的血清学调查   总被引:1,自引:0,他引:1  
用改良凝集试验(MAT)和SPA-ELISA对上海动物园珍稀野生动物的血清作了弓形虫抗体的检测。在隶属于2纲、10目、18科、37属、52种(包括亚种)的117头动物中,MAT阳性者41头(35%);ELISA检测98头动物中阳性者33头(33.7%),2种试验的阳性率无显著差异(P>0.05)。MAT的阳性率在鸟类组、灵长类组、肉食类组和草食类组中分别为11.1%(4/36)、25.0%(4/16)、69.4%(25/36)和27.6%(8/29);ELISA的阳性率分别为0.0%(0/36)、33.3%(4/12)、87.1%(27/31)和10.5%(2/19),2种试验检测不同动物组的阳性率均有非常显著差异(P<0.01)。ELISA未能在鸟类和偶蹄类动物检测到弓形虫抗体。  相似文献   

9.
用微量血凝抑制试验(HI)检测鸡减蛋综合症(EDS-76)的感染情况,结果表明,无减蛋症状鸡群平均阳性率为33.8%(97/287),HI抗体效价为1∶16-1∶128;有减蛋症状鸡群平均阳性率为92.7%(165/178),HI抗体效价多为1∶64-1∶1024,最高可达1∶16384  相似文献   

10.
23株猪瘟病毒E2基因主要抗原编码区序列差异分析   总被引:12,自引:0,他引:12  
用RT-PCR及测序获得了17株猪瘟病毒(HCV)215bp的E2基因主要抗原编码区序列,经DNAStar软件对获得的17株HCV及已发表的6株HCV毒株序列进行了比较和分析,并构建了HCV遗传发生树。结果,这23株与石门株的序列相比,所有毒株的碱基变化随机地分布于整个序列,无缺失和插入,其中变化较大的区域位于序列的3’端。23株HCV E2基因主要抗原编码区核苷酸及氨基酸同源性分别74.1%-100%、79.7%-100%,其中4株20世纪70-80年代分离毒株的核苷酸及氨基酸同源性分别为76.3%-86.2%、81.1%-87.8%,10株20世纪90年代分离毒株的核苷酸及氨基酸同源性分别为75.4%-100%、79.7%-100%。所绘制的遗传发生树分为2个组群(group),每个组群分为2个亚组群(subgroup),14株猪瘟(HC)流行毒株在2个组群中均有分布,20世纪70-80年代分离的3株(75%)在组群2,20世纪90年代分离的5株(50%)在组群1。  相似文献   

11.
迟缓爱德华氏菌胞外产物的细胞毒性和动物致病性   总被引:15,自引:0,他引:15  
分析了28株迟缓受德华氏菌(Edwardwiella tarda,Et)胞外产物(extracellular puodrcts,ECP)的致病性。将Et用覆有玻璃纸的TSA培养,18株(69.23%)的 ECP使HEp-2细胞产生病认,认圆,脱落。这种细胞毒作用能被ATCC15947株的ECP抗体作为探 针进行免疫转印分析,结果显示,不同来源的9株Et的ECP在 转印膜上出现3条共同蛋白条带,相对分子质量分别约14400、32、300、79、  相似文献   

12.
13.
迟缓爱德华氏菌的溶血特性   总被引:7,自引:0,他引:7  
分析了不同来源的28株Et的溶血特性。分别以平板法(PlateAssay,PA)、接触法(ContactHemolysis,CH)及上清法(SupernatantAsay,SA)检测Et的溶血性,阳性率依次为75%、75%、5714%。PA和CH的符合率为100%。凡SA检测为阳性的,PA、CH检测皆为阳性。培养基中加入螯合剂EDDA造成环境缺铁,结果,约3214%Et的胞外溶血素显著增加。经胰酶、热处理后,ATCC15947株溶血活性丧失,提示Et溶血素是一种不耐热蛋白样物质。  相似文献   

14.
A double-antibody sandwich enzyme-linked immunosorbent assay (ELISA) was developed for the detection and the identification of Streptococcus suis capsular types 1, 2, 1/2, 3 and 22. The specificity of this test was first evaluated using reference strains of S. suis capsular types 1 to 28 and 1/2 as well as 15 different bacterial species susceptible to be isolated from swine. The ELISA developed was very specific for capsular types 1, 3 and 22 but it could not discriminate between capsular types 2 and 1/2. In a second study, S. suis isolates from 328, 493, 368 and 76 diseased pigs were used to detect capsular types 1, 2 or 1/2, 3 and 22 respectively. The relative specificity and sensitivity varied between 98% and 100%. The ELISA results were in excellent agreement with the standard techniques (biochemical tests, coagglutination and capsular reaction tests) in detecting both positive and negative strains. Kappa values were 0.80, 0.99, 0.97 and 1.00 for detecting S. suis capsular types 1, 2 or 1/2, 3, and 22 respectively. To evaluate the relative-sensitivity of the test, primary cultures from 73 diseased pigs and tissue samples from 67 diseased pigs were used directly for detecting these capsular types. With primary cultures, the relative specificity and sensitivity (95.9% and 91.6% respectively) remained high and the test was very suitable (Kappa = 0.87). The ELISA using tissue samples gave a good specificity (97.6%), a moderate sensitivity (62.5%) and a low agreement with standard tests (Kappa = 0.64).(ABSTRACT TRUNCATED AT 250 WORDS)  相似文献   

15.
A polymerase chain reaction (PCR) assay was developed for the generic detection of Salmonella sp. and the identification of S. Enteritidis (SE), S. Gallinarum (SG), S. Pullorum (SP) and S. Typhimurium (ST) in material collected in the field from poultry. The specificity and sensitivity of the assay combined with Rappaport-Vassiliadis selective enrichment broth (PCR-RV) were determined, and field samples were analyzed to verify the validity of the method application. Specificity of the assay was tested using 29 SE, 11 SG, 10 ST and 10 SP strains, along with 75 strains of 28 other Salmonella serovars and 21 strains of other bacterial genera. The assay was 100% specific for Salmonella detection and ST identification. The primer pair for SE, SG and SP also detected S. Berta. PCR detection limits for Salmonella at the genus level were 2 ST, 8 SE, 1.1x10(3) SG and 1.8x10(5) SP cells. At the serovar level, detection limits were 7 ST, 1.2x10(3) SE, 4.4x10(7) SG and 1.8x10(6) SP cells. At the genus level, PCR-RV detected approximately 128% more positive field samples than the standard microbiological techniques and results were ready in 48h instead of 7 days. PCR-RV method is diagnostic of Salmonella at the genus level and ST at the serovar level, although other tests are needed to identify SE, SG and SP to serovar level.  相似文献   

16.
实时荧光定量RT-PCR检测牛库布病毒   总被引:1,自引:1,他引:0  
牛库布病毒(bovine kobuvirus,BKoV)可能是一个腹泻相关病原,目前还没有检测该病毒的实时荧光定量RT-PCR方法的报道。依据BKoV 3D核苷酸序列,采用Primer 6.0设计了一对引物,通过优化反应条件和反应体系,成功建立了检测BKoV的TB Green染料法实时荧光定量RT-PCR方法。结果显示此方法在4.86×108~4.86×102 copies·μL-1病毒浓度之间具有良好的线性关系,相关系数为0.999 5,扩增效率为92.75%;此方法只特异性检测BKoV,而与其他常见的腹泻病原没有交叉反应;组内及组间的变异系数均小于2%;最低检测下限为4.86×102 copies·μL-1。本研究所建的实时荧光定量RT-PCR方法对奶牛和牦牛腹泻粪便样本中BKoV的检出率明显高于已报道的3篇有关RT-PCR方法。对2018年5月-2019年5月采集自青藏高原地区(青海、西藏和四川藏区)的131份牦牛腹泻粪便样本进行检测,结果显示BKoV检出率为31.30%。首次成功建立了检测BKoV的实时荧光定量RT-PCR方法,具有良好的特异性和稳定性,且灵敏度高,为BKoV的检测及流行病学调查提供了一种新的技术手段;并且首次证实BKoV在牦牛中的流行,为牦牛腹泻的防控问题提供了参考。  相似文献   

17.
为建立快速检测鹿血中副结核分枝杆菌(Mycobacterium avium subsp. paratuberculosis,MAP)的荧光定量PCR方法,本研究根据GenBank中登录的MAPf57基因序列设计并合成引物及探针,并检测该方法的特异性和敏感性。试验结果显示,该方法具有良好的特异性,对MAP的检测灵敏度可以达到单个菌细胞。对长春地区采集的549份血清样品进行检测,结果显示阳性血清101份,阳性率达到18.4%。本研究结果表明,荧光定量PCR用于动物性产品MAP的检测具有快速、准确的特点。  相似文献   

18.
应用以葡萄球菌A蛋白协同凝集试验(SPA-COA)为主要检测手段的鱼病诊断箱,于1995 ̄1997年在吉林,辽宁等7省进行现场检测试验。结果,61个渔场11个品种的516尾病鱼有菌生长的285尾,经SPA-COA鉴定阳性数为239尾,占有菌生长的83.86%,12个渔场5个品种1216尾假定健康鱼有菌生长的为28尾,经SPA-COA鉴定阳性数为22尾,占有菌生长的78.57%,认为本诊断箱适用于基  相似文献   

19.
A sandwich-dot enzyme-linked immunosorbent assay (dot ELISA) was developed for the detection of canine distemper virus (CDV). In 56 dogs suspected to have CD the rates of detection of CDV antigen in samples of blood lymphocytes and palpebral conjunctiva by dot ELISA and ELISA were, respectively, 91% (49/54) and 81% (44/54) for the lymphocyte samples and 88% (28/32) and 75% (24/32) for the conjunctival samples. The CDV detection limits were 10 ng/50 μL for dot ELISA and 40 ng/50 μL for ELISA. The reliability of dot ELISA relative to electron microscopy was 96% with 22 samples: all 21 samples in which CDV particles were observed by electron microscopy yielded positive results with dot ELISA; the single sample in which particles were not observed yielded false-positive results with dot ELISA. The results indicate that the dot ELISA developed can serve as a reliable rapid diagnostic test in suspected cases of CD and also be useful for epidemiologic surveillance of the disease.  相似文献   

20.
OBJECTIVE: To develop a polymerase chain reaction (PCR) assay that detects and differentiates the Ohio strain of Haemobartonella felis (H. felis-OH) and the California strain of H. felis (H. felis-CA) and to apply the assay to blood samples from cats with and without suspected haemobartonellosis (suspect and control cats, respectively). SAMPLE POPULATION: 220 blood samples were examined; 82 were from suspect cats, and 138 were from control cats. PROCEDURE: A PCR assay was designed to detect and differentiate H. felis-OH and H. felis-CA. RESULTS: On the basis of PCR assay results, the overall prevalence of H. felis infection was 19.5% (43/220). Suspect cats (28.0%; 23/82) were significantly more likely than control cats (14.5%; 20/138) to be H. felis infected. Significantly greater numbers of suspect cats were H. felis-OH infected (12.2%, 9/82) or H. felis-OH and H. felis-CA infected (4.9%, 4/82) than control cats (0% [0/138] and 0.7% [1/138], respectively). Significantly more anemic cats were H. felis-OH infected (14.3%; 4/28) or H. felis-OH and H. felis-CA infected (7.1%; 2/28) than nonanemic cats (2.3% [3/128] and 0.8% [1/128], respectively). The PCR assay was more accurate than cytologic examination for detection of H. felis. CONCLUSIONS AND CLINICAL RELEVANCE: Haemobartonella felis infections are more common in cats than previously recognized. Haemobartonella felis-OH is apparently more pathogenic than H. felis-CA. The PCR assay is more accurate than cytologic examination for detection of H. felis infection and is an effective clinical tool for the detection and differentiation of both H. felis strains known to infect cats.  相似文献   

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