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首页> 外文期刊>Global Veterinaria >Amos PCR as a Rapid Screening Method for Differentiation of Infected and Vaccinated Cattle and Sheep with Brucellosis
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Amos PCR as a Rapid Screening Method for Differentiation of Infected and Vaccinated Cattle and Sheep with Brucellosis

机译：Amos PCR作为一种快速筛选方法，用于鉴别感染和接种牛布鲁氏菌病的牛和羊

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This study was carried out on 55 animals (5 vaccinated cattle with S19 vaccine, 5 vaccinated cattlewith RB51 vaccine, 5 vaccinated Sheep with Riv 1 vaccine, 20 cattle from suspected to be infected or had ahistory of brucellosis and 20 sheep from mobile flocks of known history of brucellosis). The results ofserological tests which were applied on serum samples, revealed out of 5/5 vaccinated cattle with S19(100%),(100%) and (100.0%) were positive by RBPT, TAT and Riv.T respectively so vaccinated animals tend to yieldpersistent post-vaccinal immune responses. While out of 5/5 vaccinated cattle with RB51 were negative by thesame serological tests as RB51 a rough strain of Brucella can not detected by these tests. But out of 20 cattlefrom infected farm with Brucella 5/20(25%), 5/20(25%) and 5/20(25%) were positive by RBPT, TAT and Riv.Trespectively. Out of 5 vaccinated Sheep with Riv 1 vaccine 5/5(100%), 5/5(100%) and 5/5(100%) were positiveby RBPT, TAT and Riv.T respectively while out of 20 sheep from infected mobile flocks 4/20 (20%), 4/20(20%)and 4/20(20%) were positive by RBPT, TAT and Riv.T respectively. Out of 5 milk samples were collected fromseropositive naturally infected cattle and 4 milk samples from seropositive naturally infected sheep werepositive PCR amplifies, 731 bp fragment indicated (B. melitensis biovar 3), which considered the most prevalentstrain in Egypt. AMOS PCR results were recorded by electrophoretic mobilitis in agarose gels and photographicmethods showed two products (498 and 364 bp) plus 178-bp were amplified from RB51 vaccinated cattle butonly one product (498 bp) was amplified from the S19 vaccinated cattle. Amplified fragments obtained fromvaccinated sheep with Rev 1 vaccine produce 2 bands, 282 bp and 238 bp, the product obtained from digestionof the amplified fragment. In contrast naturally infected sheep with B.melitensis field isolate (B. melitensisbiovar3) produced only one band a 238-bp fragment. AMOS PCR was shown to be a valuable tool and a rapidscreening test for differentiating the vaccinated animals with S19, RB51 or Riv 1 from infected one withB. melitensis biovar 3(field strains of Brucella) which means that Brucellosis eradication program could reliablyuse the Brucella AMOS PCR without any supplement to other diagnostic and epidemiological data to releasesale animals from quarantine before the conventional identification methods are completed.
机译：该研究针对55只动物进行了研究（5头接种S19疫苗的牛，5头接种RB51疫苗的牛，5头Riv 1疫苗的牛，20头怀疑感染或有布鲁氏菌病史的牛和20头来自已知家禽的羊布鲁氏菌病的历史）。对血清样本进行的血清学测试结果表明，在5/5接种S19的牛中，RBPT，TAT和Riv.T分别为S19（100％），（100％）和（100.0％）阳性，因此接种动物倾向于产生持久的疫苗后免疫反应。尽管在5/5接种RB51的牛中，血清学检测结果与RB51相同，但布鲁氏菌的粗菌株在这些检测中无法检测到。但是在感染布鲁氏菌5/20（25％）的农场的20头牛中，RBPT，TAT和Riv分别为5/20（25％）和5/20（25％）阳性。在接受Riv 1疫苗的5只接种绵羊中，RBPT，TAT和Riv.T分别对5/5（100％），5/5（100％）和5/5（100％）呈阳性，而在20只绵羊中被感染RBPT，TAT和Riv.T分别对4/20（20％），4/20（20％）和4/20（20％）鸡群呈阳性。从血清阳性的自然感染的牛中采集了5份牛奶样品，对血清阳性的自然感染的绵羊中的4份牛奶样品进行了PCR扩增，显示了731 bp的片段（B. melitensis biovar 3），这被认为是埃及最流行的菌株。在琼脂糖凝胶上通过电泳迁移率法记录了AMOS PCR结果，照相法显示从RB51接种牛中扩增出两种产物（498和364 bp），外加178 bp，而从S19接种牛中扩增出仅一种产物（498 bp）。从接种有Rev 1疫苗的羊中获得的扩增片段产生2条带，分别为282 bp和238 bp，这是消化该扩增片段获得的产物。相比之下，自然感染的带有B.melitensis野外分离株（B. melitensisbiovar3）的绵羊只产生一条238 bp的片段。已证明，AMOS PCR是区分S19，RB51或Riv 1接种动物与B感染动物的一种有价值的工具和快速筛选测试。 melitensis biovar 3（布鲁氏菌的野外菌株），这意味着消除布鲁氏菌病程序可以可靠地使用布鲁氏菌AMOS PCR，而无需对其他诊断和流行病学数据进行任何补充即可在常规鉴定方法完成之前将动物从隔离区中释放出来。

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