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《畜牧兽医学报》2016,(4)
本研究的目的是探索副结核分枝杆菌特异性蛋白MAP0862在牛副结核病血清学诊断中的作用,建立牛副结核病特异性ELISA诊断方法。将通过原核表达获得的副结核特异性蛋白MAP0862纯化并定量后作为包被抗原,经过一系列条件优化后初步建立了牛副结核病间接ELISA诊断方法。使用牛副结核阳性血清、牛布病阳性血清、牛结核阳性血清、卡介苗免疫牛血清、牛大肠杆菌阳性血清以及健康阴性牛血清对该方法的验证表明其具有良好的特异性。使用该方法对300份临床血清进行检测,结果表明该方法与IDEXX副结核检测试剂盒的符合率为94.3%。基于副结核特异性蛋白质MAP0862建立的间接ELISA诊断方法能有效检测牛副结核病。 相似文献
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牛副结核病(bovine paratuberculosis)是由副结核分支杆菌引起的以牛慢性增生性、顽固性肠炎和进行性消瘦为特征的人畜共患传染病。其临床症状与病理变化是顽固性腹泻、渐进性消瘦、慢性肉芽肿回肠炎,肠粘膜增厚并形成皱褶。除常规的临床诊断外,牛副结核病的确诊依赖于实验室诊断方法,主要有病原学诊断、血清学诊断和分子生物学诊断等方法。牛副结核病给畜牧业造成很大的损失。由于本病潜伏期长,传播快,加之人工培养副结核分枝杆菌难度又高,因此加强对该病的病原学、流行病学特征的认识,加快实验室诊断方法的研究对其综合防控工作具有十分重要的意义。作者主要介绍了目前国内外牛副结核病流行病学特征和诊断方法的研究进展,并比较了各种方法的优点和缺点,为牛副结核病的临床快速诊断和深入研究提供科学依据。 相似文献
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《养殖与饲料．饲料世界》2021,(7)
牛副结核病是一种由副结核分枝杆菌引起的慢性传染病,主要表现为顽固性腹泻、渐进性消瘦等,病牛和副结核分枝杆菌携带牛是该病的主要传染源。本文介绍了该病的病原学、流行病学、临床症状、病理变化、诊断方法和防控策略。该病目前没有特效药物来治疗,重在预防,主要通过隔离疑似病牛、加强检疫、加强饲养管理等措施综合防控。 相似文献
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为了解湖南省规模牛场牛病毒性腹泻、牛传染性鼻气管炎、牛结核病、牛副结核病4种疫病的流行状况,采用ELISA方法,对全省未免疫以上4种疫病的规模牛场开展了血清学调查。结果显示：湖南省规模牛场的牛病毒性腹泻、牛传染性鼻气管炎、牛结核病、牛副结核病的场阳性率分别为51.35%、74.32%、13.64%、22.73%,个体阳性率分别为23.54%、45.45%、3.11%、4.67%。全省14个市州规模牛场均检出牛病毒性腹泻与牛传染性鼻气管炎病原抗体阳性,4个市州检出牛分枝杆菌抗体阳性,7个市州检出副结核分枝杆菌抗体阳性;不同规模牛场4种疫病的抗体阳性率存在一定差异,大规模场偏高;多重感染占比为45.45%。结果表明：牛病毒性腹泻和牛传染性鼻气管炎在湖南省规模牛场流行较为普遍、严重,部分地区还存在牛结核病、牛副结核病流行,而在大规模牛场4种疫病流行更为严重,各规模牛场4种疫病病原的多重感染情况也较为普遍。因此,湖南省规模牛场4种疫病防控形势严峻,需加强防控。本调查为湖南省此类疫病防控提供了数据参考。 相似文献
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副结核病是由禽分枝杆菌副结核亚种引起的慢性疾病,会感染多种反刍动物.副结核病的流行病学十分复杂,该病在牛中的临床表现和经济影响可能因畜群管理、年龄、感染菌量和疾病流行情况等因素而异.此外,在控制牛副结核病方面还面临着相当大的挑战,如缺乏准确可靠的诊断检测方法.尽管如此,全球仍致力于控制牛副结核病,因为该病对养牛羊可造成... 相似文献
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牛副结核检疫中的非特异性干扰 总被引：2，自引：0，他引：2
给犊牛接种各种分枝杆菌，然后以副结核ＰＰＤ进行变态反应试验，观察各分枝杆菌感染对副结核变态反应的干扰；从各种分枝杆菌感染牛群采取血清进行副结核ＥＬＩＳＡ检测，以观察各分枝杆菌血清对牛副结核ＥＬＩＳＡ的干扰。结果：多种分枝杆菌，特别是鸟胞内分枝杆菌、瘰疬分枝杆菌的感染可干扰副结核变态反应；部分结核牛血清干扰副结核ＥＬＩＳＡ的检测 相似文献
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G Holstad óG Sigureardóttir AK Storset J Tharaldsen O Nyberg J Sch?nheit B Dj?nne 《Acta veterinaria Scandinavica》2005,46(1):45-56
The Norwegian surveillance and control programme for paratuberculosis revealed 8 seroreactors in a single dairy cattle herd that had no clinical signs of Mycobacterium avium subsp. paratuberculosis (M. a. paratuberculosis) infection. Paratuberculosis had been a clinical problem in goats several years previously in this herd. All 45 cattle were culled and a thorough investigation of the infection status was conducted by the use of interferon-γ (IFN-γ) immunoassay, measurement of antibodies, and pathological and bacteriological examination.In the IFN-γ immunoassay, 9 animals gave positive results, and 13 were weakly positive, while 19 animals were negative. In the serological test,10 animals showed positive reactions, and 5 were doubtful, while 30 animals gave negative reactions. There appeared to be a weak trend toward younger animals having raised IFN-γ and older animals having raised serological tests. Histopathological lesions compatible with paratuberculosis were diagnosed in 4 animals aged between 4 and 9 years. Three of these animals had positive serological reaction and one animal gave also positive results in the IFN-γ immunoassay. Infection was confirmed by isolation of M. a. paratuberculosis from 2 of these 4 animals. One single bacterial isolate examined by restriction fragment length polymorphism (RFLP) had the same profile, B-C1, as a strain that had been isolated from a goat at the same farm several years previously.Despite many animals being positive in one or both of the immunological tests, indicative of a heavily infected herd, none of the animals showed clinical signs and only one cow was shown to be shedding bacteria. A cross-reaction with other mycobacteria might have caused some of the immunoreactions in these animals. It is also possible that the Norwegian red cattle breed is resistant to clinical infection with M. a. paratuberculosis. 相似文献
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Nagaraja R. Thirumalapura Willard Feria Eric Hue Corey Zellers Deepanker Tewari 《Journal of veterinary diagnostic investigation》2021,33(2):375
Johne’s disease (paratuberculosis) is an economically important disease of cattle worldwide. The disease is caused by Mycobacterium avium subsp. paratuberculosis (MAP), a fastidious gram-positive bacterium. PCR is increasingly used in diagnostic laboratories for the detection of MAP in fecal samples given the rapid test turnaround time and sensitivity and specificity comparable to fecal culture. However, efficient extraction of DNA for sensitive detection of MAP by PCR is affected by the complex lipid-rich cell wall of MAP and the presence of PCR inhibitors in feces. We evaluated a high-throughput nucleic acid extraction method (MagMAX core nucleic acid purification kit with mechanical lysis module) in conjunction with an hspX gene PCR for the detection of MAP from bovine fecal samples, which resulted in correct identification of all negative (13 of 13) and positive (35 of 35) proficiency test samples obtained from the National Veterinary Services Laboratories. In addition, all 6 negative and 50 of 51 positive diagnostic specimens tested were categorized correctly. 相似文献
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羊副结核病是由副结核分枝杆菌引起的一种以顽固性腹泻、肠粘膜显著增厚为主要临床特征的慢性传染病,多呈现隐形感染,该病潜伏期长,传播迅速,呈全球性分布,对畜牧业的危害巨大。本文立足羊副结核病研究进展的实际情况,梳理总结了目前学术界对羊副结核病病原学、流行病学、致病机制、临床诊断和免疫综合防控等方面的研究讨论,为羊副结核病的诊断与防控提供了理论基础。在此基础上提出防控羊副结核病政府需制定有效政策,养殖场需加强饲养管理,形成科学有效的防控系统,为我国畜牧业的可持续健康发展提供了理论支撑。 相似文献
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Gwozdz JM Thompson KG Murray A Reichel MP Manktelow BW West DM 《Australian veterinary journal》2000,78(11):779-783
OBJECTIVE: To compare the diagnostic performance of a complement fixation test, an agar gel immunodiffusion test, an enzyme-linked immunosorbent assay, and a whole-blood interferon-gamma assay for paratuberculosis in 14 sheep experimentally infected with Mycobacterium avium subsp paratuberculosis. EXPERIMENTAL DESIGN: Longitudinal study. RESULTS: The IFN-gamma assay detected more experimentally infected sheep, and earlier, than any of the serological tests. None of the antibody assays was able to detect all sheep with histologically confirmed paratuberculosis. CONCLUSIONS: The superior performance of the IFN-gamma assay in detecting infected sheep in this small experimental population warrants its further evaluation in a larger population of sheep naturally exposed to M a paratuberculosis. 相似文献
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OBJECTIVE: To determine the sensitivity and specificity of an absorbed ELISA and an AGID test for the detection of clinical and subclinical paratuberculosis in sheep. DESIGN: By testing a panel of sera from 1257 Australian Merino and crossbred sheep greater than 1 year of age, of which 1137 sheep were not infected with Mycobacterium avium subsp paratuberculosis and 120 sheep had paratuberculosis. PROCEDURE: Sera were collected from 457 sheep in Victoria and 800 sheep in Western Australia. Presence of M a paratuberculosis infection in Victorian sheep was determined by histological examination of intestinal tissues, whereas sheep from Western Australia were presumed to be free of Johne's disease. The ability of an absorbed ELISA to discriminate between infected and uninfected sheep was described by test sensitivity and specificity, the distribution of ELISA OD, and the area under a receiver operating characteristic curve. RESULTS: The absorbed ELISA had a specificity of 98.2 to 99.5% (CI) and a sensitivity of 35 to 54% (CI). In sheep from infected flocks in Victoria, the AGID test had a specificity of 99 to 100% (CI) and a sensitivity of 38 to 56% (CI). The sensitivity of serological tests was higher in sheep with a body condition representative of the lower quintile of their flock of origin. CONCLUSION: The AGID test and absorbed ELISA are useful tests for the detection of ovine paratuberculosis. Although the tests had a similar accuracy, they detected different subpopulations of infected sheep with only moderate overlap. The AGID test had a higher specificity than the absorbed ELISA. 相似文献
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OBJECTIVES: To evaluate additional restriction enzymes for IS900 RFLP of Mycobacterium avium subsp paratuberculosis and examine the genetic diversity among Australian isolates for epidemiological studies of Johne's disease. DESIGN AND PROCEDURE: Seventy-one isolates of M paratuberculosis from cattle, sheep, goat, alpaca and rhinoceros in six Australian States and the Northern Territory, reference strains and reference DNA from previously characterised strains were tested for genetic variation. Bst EII, Pvu II and Pst I restriction enzymes were used, and four others (Bam HI, Alu I, Xho I and Dra I) were assessed for their ability to detect polymorphisms. Multiple isolates from some animals were tested. RESULTS: Bam HI, was the most effective enzyme for identifying polymorphisms (12 types), followed by Bst EII (11 types). Both Pvu II and Pst I were relatively ineffectual. Fifteen different types were identified, 12 in clinical isolates. Most isolates were cattle (C) strains and fell into the C1 (n = 28) and C3 (n = 32) groupings. All isolates from alpaca were type C1, and bovine isolates were commonly C1 (n = 15) or C3 (n = 28). All of the sheep were infected with sheep (S) strains; no S strains were identified in cattle. Two of six isolates from one animal had single band differences. CONCLUSION: The epidemiological features of M paratuberculosis in Australia are similar to those reported in New Zealand, where cattle and sheep are commonly infected with different strains. However, because of the lack of polymorphism identified within the major groups, it is unlikely that DNA fingerprinting will have a significant role in epidemiological studies of Johne's disease, unless an unusual strain in being studied. 相似文献
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Kun Taek Park Jongsam Ahn William C. Davis Hye Cheong Koo Nam Hoon Kwon Woo Kyung Jung Jun Man Kim Soon Keun Hong Yong Ho Park 《Journal of veterinary science (Suw?n-si, Korea)》2006,7(4):349-354
Paratuberculosis (PTB) is a major disease problem worldwide, and causes major economic losses in the dairy industry. Although PTB has been reported in Korea, no studies have been conducted to determine its prevalence and no program has been developed to control the disease. In this study, the sera of beef (n = 1,056) and dairy cattle (n = 1,105) from all provinces in Korea were tested to determine the prevalence of PTB using two different ELISA: an ''in house'' modified absorbed ELISA (P-ELISA) based on sonicated antigen from Mycobacterium avium subsp. paratuberculosis ATCC 19698, and a commercial ELISA (C-ELISA). Receiver operating characteristic analysis was used to determine the cutoff point for P-ELISA. Based on C-ELISA results, the area under the curve for P-ELISA was 0.913 (95% CI, 0.883 to 0.943). Using a cutoff point of 0.100, P-ELISA showed a sensitivity of 62.0% and a specificity of 93.7%. The kappa value and the percent agreement between the two ELISAs were 0.322 and 92.5%, respectively. Both ELISAs showed a significant correlation between age and seropositivity (p < 0.01). According to C-ELISA, 71 of 2,161 sera (3.3%, 95 CI, 2.6% to 4.1%) were test-positive. The national true prevalence of PTB was estimated to be 7.1%. The findings suggest that a control program should be implemented to limit the spread of this disease, and that P-ELISA could be used as a screening test that produces results similar to C-ELISA. 相似文献
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Miguel Salgado Cristobal Verdugo Cord Heuer Pedro Castillo Patricia Zamorano 《Journal of veterinary science (Suw?n-si, Korea)》2014,15(2):233-239
PCR is a highly accurate technique for confirming the presence of Mycobacterium avium subsp. paratuberculosis (Map) in broth culture. In this study, a simple, efficient, and low-cost method of harvesting DNA from Map cultured in liquid medium was developed. The proposed protocol (Universidad Austral de Chile [UACH]) was evaluated by comparing its performance to that of two traditional techniques (a QIAamp DNA Stool Mini Kit and cethyltrimethylammonium bromide [CTAB] method). The results were statistically assessed by agreement analysis for which differences in the number of cycles to positive (CP) were compared by Student''s t-test for paired samples and regression analysis. Twelve out of 104 fecal pools cultured were positive. The final PCR results for 11 samples analyzed with the QIAamp and UACH methods or ones examined with the QIAamp and CTAB methods were in agreement. Complete (100%) agreement was observed between data from the CTAB and UACH methods. CP values for the UACH and CTAB techniques were not significantly different, while the UACH method yielded significantly lower CP values compared to the QIAamp kit. The proposed extraction method combines reliability and efficiency with simplicity and lower cost. 相似文献
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为协助新疆某副结核病严重流行的奶牛场抑制该病的流行,2014-2015年间采用副结核分枝杆菌(MAP)间接ELISA抗体检测试剂盒对该牛场的965头牛进行了4次血清抗体检测,并通过淘汰部分阳性牛控制该病的流行。结果显示,在两年内累计检测出297头阳性牛,检出率为30.78%(297/965)。首次检测发现104头阳性牛,检出率为13.51%(104/770)。淘汰处理后,间隔3个月再次检出2.91%(19/652)的阳性牛,再次淘汰阳性牛后进行了两次监测,其中间隔11个月时检测阳性率为7.55%(41/543),间隔17个月时检出率为33.33%(171/513)。同时,检测结果显示:抗体阳性牛的检出率随年龄的增加而呈现上升趋势,在4~5岁检出率最高,而0.5~1和7~8岁牛中检出率较低。统计分析发现,在副结核病严重流行的奶牛场开展该病的控制或净化时,检测和主动淘汰的间隔期设置为3个月时的控制效果优于11个月和17个月(P<0.001)。虽本次处置未达到预期目标,但根据检测数据和处置措施效果,结合现有的研究数据,初步制订了牛群副结核病流行程度的划分标准和防控、净化方案。本研究将为牛副结核病的防控提供案例借鉴和防控方案。 相似文献
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Gwozdz JM Thompson KG Murray A West DM Manktelow BW 《Australian veterinary journal》2000,78(9):622-624
OBJECTIVE: To evaluate a polymerase chain reaction-based assay for the detection of Mycobacterium avium subsp paratuberculosis in blood and liver biopsies from subclinically infected sheep. STUDY DESIGN: A direct PCR assay for the detection of M a paratuberculosis was applied to liver biopsy specimens and to samples of blood that were sequentially collected over 53 weeks from 14 sheep infected experimentally with the organism. RESULTS: Of 117 blood samples from the 14 experimentally infected sheep, two tested positive in the PCR assay. Both positive results were obtained in two subclinically infected sheep that had paratuberculosis later confirmed by histological examination at necropsy. However, the assay failed to detect the target DNA in samples of blood from five other sheep with histologically confirmed paratuberculosis. Similarly, the PCR assay on liver biopsy specimens collected 32 weeks after administration of M a paratuberculosis gave only two positive results, both of which were obtained in sheep with histological evidence of paratuberculosis. CONCLUSIONS: The PCR assay on blood and liver biopsies does not provide a useful method for the diagnosis of M a paratuberculosis infection in subclinically infected sheep. 相似文献