Isolation and molecular detection of duck plague virus for the development of vaccine seed

Sabiha Sultana Soma, K. Nazir, M. T. Rahman, Md. Mizanur Rahman, M. Ara, R. Sultana, M. R. Haydar, M. Siddique, M. Rahman
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引用次数: 3

Abstract

The present study was carried out for the isolation and molecular detection of duck plague virus (DPV) for the development of inactivated vaccine seed from the local outbreaks. A total of 12 suspected dead duck samples were collected from commercial farms and local market at Sunamganj, Netrokona and Mymensingh districts. Then, the samples were processed and prepared inocula were inoculated into 9-12 days old duck embryonated eggs. In duck embryonated eggs, several passages (3-4) were performed before infection into DEF cell culture. Presence of viral DNA was confirmed by PCR using the primer for DNA polymerase gene. After PCR confirmation, virus cultured in DEF cell was used for the preparation of formalin (0.12%) inactivated and oil based adjuvanted vaccine and was experimentally injected to 18 ducklings and 5 were kept as control. TCID50 of the selected virus for vaccine preparation was 108.70/ml. The mean passive haemagglutination assay (PHA) titre of sera of samples at 0 days, 7 days and 14 days post vaccination were 4.0±0, 14.22±1.78 and 44.44±4.4, respectively, which indicated significant (p<0.01) increase of antibody titre. Embryonated duck eggs and DEF cell culture are effective for virus isolation and on the basis of PHA test, it could also be suggested that the experimentally developed DP vaccine can be used successfully for the prevention of DP in Bangladesh. Asian Australas. J. Biosci. Biotechnol. 2018, 3 (1), 78-85
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鸭瘟病毒的分离与分子检测用于疫苗种子的研制
本研究旨在对鸭瘟病毒(DPV)进行分离和分子检测,以开发鸭瘟灭活疫苗种子。在Sunamganj、Netrokona和Mymensingh地区的商业农场和当地市场共采集了12只疑似死鸭样本。将制备好的疫苗接种于9 ~ 12日龄的鸭胚蛋中。在鸭胚蛋中,在感染DEF细胞培养前进行了几次传代(3-4次)。利用DNA聚合酶基因引物进行PCR,证实病毒DNA的存在。经PCR确认后,用DEF细胞培养的病毒制备福尔马林(0.12%)灭活疫苗和油基佐剂疫苗,实验注射18只雏鸭,留5只作对照。所选疫苗制备病毒的TCID50为108.70/ml。接种后第0天、第7天和第14天,血清被动血凝试验(PHA)滴度分别为4.0±0、14.22±1.78和44.44±4.4,抗体滴度均显著(p<0.01)升高。鸭蛋胚和DEF细胞培养可有效分离病毒,PHA试验也表明,实验研制的DP疫苗可成功用于孟加拉国的DP预防。亚洲Australas。j . Biosci。生物技术,2018,3 (1),78-85
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