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首页> 外文期刊>Journal of Veterinary Advances >Isolation and Molecular Characterization of Foot and Mouth Disease SAT2 Virus during Outbreak 2012 in Egypt
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Isolation and Molecular Characterization of Foot and Mouth Disease SAT2 Virus during Outbreak 2012 in Egypt

机译：埃及爆发2012年口蹄疫SAT2病毒的分离和分子特征

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During April and May 2012, six outbreaks of FMD type SAT 2 were reported in Egyptian governorates: EL- Gharbiya (Kafr Qeretna, Al – Mahala Alkobra, Ebshaway Almalak, Qutoor), local cattle, buffaloes and dairy farms in Kafr El-Shaykh (Alsalmya, Fouah), AL - Minya (Village 3, Samalout, Mansafees, Abo Querkas), Dumyat (Ezbet 20, Kafr saad), ALexandria (Iraq Village, Al Aamerya), EL minufya (Monshaat Gerges, Ashmoun, Shatanoof, Ezbet Algalayla, Berket Alsabaa, Mahalet Sabak),Luxor (Al Gorna) and Al- Sharqiya (Al -Telleen, Menya Al kamh) Governorates. Suspected FMD virus samples (22 Tongue epithelium (TE), 12 vesicular fluid samples (VF)), in addition to, 10 cell culture grown virus of tongue epithelium origin, 6 cell culture grown virus of vesicular fluid origin. The assays used in this study involved CFT and Indirect ELISA typing kits for typing of FMDV virus samples; 3ABC-FMD for differentiation between vaccinated and infected animals in any revealed protective immune response in the examined sera from Egypt 2012 outbreak. PCR as an advanced confirmatory technique was done before submitting FMDV suspected clinical samples for Reference Laboratory of Foot and Mouth Disease in Pirbright, United Kingdom. Samples were isolated on primary bovine kidney and Baby Hamster kidney cell cultures, where all samples gave cytopathic effect. Also, all virus samples inoculated in swiss albino suckling baby mice showed pathognomonic paralysis effect of the virus in hind limbs of mice. Serum samples collected from suspected infected cattle were examined using FMD-3ABC. All positive samples to 3ABC antibodies indicated that the animals were infected by FMDV. FMD antibody detection revealed infection of 62%, 100%, 61%, 37.5%, 45.5% , 72.7% and 66.6% of sera collected from Al-Gharbiya, Kafr El-Shaykh, Dumyat, Alexandria, Al-Monufya, Luxor and Al-Sharqiya respectively. The obtained results of 3ABC antibodies were subsequently confirmed by Indirect ELISA Kit, which revealed that the causative agent of the outbreak was FMDV SAT2. The prefinal confirmatory technique was the Reverse transcription polymerase chain reaction (RT-PCR), which was done to detect clinically suspicious samples using target primers. Also, DNA sequencing of the current FMDV strain was performed and revealed that there was no significant divergence among the isolated strains as the divergence was 0.3 to 2.2% among them, and also there was no significant divergence between the isolated strains and FMDV SAT2/EGY/5/2012 and SAT2/EGY/15/2012 The closest percentage of identity of the isolated strains in comparison with reference strains was 89.4% with FMDV SAT2/LIB/2003. For official confirmation, the isolated viruses were sent to the Reference Laboratory for Foot and Mouth Disease in Pirbright, United Kingdom. Isolation of the causative agent is the first step for diagnosis of the disease. Finally, the PCR product was sequenced and analyzed using the gene bank data to confirm that the new out
机译：在2012年4月至2012年5月期间，埃及各省报告了6次FMD SAT 2爆发：EL- Gharbiya（Kafr Qeretna，Al – Mahala Alkobra，Ebshaway Almalak，Qutoor），Kafr El-Shaykh（当地人），牛，水牛和奶牛场（ Alsalmya，Fouah），AL-Minya（3村，Samalout，Mansafees，Abo Querkas），Dumyat（Ezbet 20，Kafr saad），ALexandria（伊拉克伊拉克村，Al Aamerya），EL minufya（Monshaat Gerges，Ashmoun，Shatanoof，Ezbet Algalayla ，Berket Alsabaa，Mahalet Sabak），Luxor（Al Gorna）和Al-Sharqiya（Al-Telleen，Menya Al kamh）省。疑似FMD病毒样品（22个舌头上皮（TE），12个囊泡液样品（VF）），另外还有10个舌头上皮细胞培养生长病毒，6个舌头上皮细胞培养生长病毒。本研究中使用的测定方法包括CFT和间接ELISA打字试剂盒，用于FMDV病毒样品的打字。 3ABC-FMD用于区分埃及2012爆发的受检血清中任何已显示的保护性免疫应答中的接种动物和感染动物。在将FMDV疑似临床样品提交给英国Pirbright的口蹄疫参考实验室之前，已进行了PCR作为一种先进的确认技术。在原发性牛肾和小仓鼠肾细胞培养物中分离样品，所有样品均具有细胞病变作用。而且，在瑞士的白化病乳宝宝小鼠中接种的所有病毒样品在小鼠的后肢中显示出该病毒的致病性麻痹作用。使用FMD-3ABC检查从疑似感染牛身上收集的血清样本。 3ABC抗体的所有阳性样品均表明动物被FMDV感染。 FMD抗体检测显示感染了Al-Gharbiya，Kafr El-Shaykh，Dumyat，Alexandria，Al-Monufya，Luxor和Al血清的62％，100％，61％，37.5％，45.5％，72.7％和66.6％ -沙吉亚分别。随后通过间接ELISA试剂盒确认了获得的3ABC抗体结果，该结果表明暴发的病原体是FMDV SAT2。最终的确认技术是逆转录聚合酶链反应（RT-PCR），该技术可使用目标引物检测临床可疑样品。另外，对当前的FMDV菌株进行了DNA测序，结果表明，分离菌株之间没有显着差异，因为它们之间的差异为0.3％至2.2％，分离菌株与FMDV SAT2 / EGY之间也没有明显差异。 / 5/2012和SAT2 / EGY / 15/2012与FMDV SAT2 / LIB / 2003相比，分离菌株与参考菌株的最接近同一性百分比为89.4％。为了得到官方确认，已将分离出的病毒发送至英国Pirbright的口蹄疫参考实验室。隔离病原体是诊断该疾病的第一步。最后，使用基因库数据对PCR产物进行测序和分析，以确认新的

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