Rahmahani Jola, Zahro Aisyah Nikmatuz, Rahmawati Indah Laili, Putih Nurvita, Wulandari Innah, Abdul-Rantam Fedik
Laboratory of Virology and Immunology, Department of Microbiology, Faculty of Veterinary Medicine, Airlangga University, Indonesia, 60115.
Bachelor of Veterinary Science, Faculty of Veterinary Medicine, Airlangga University, Indonesia, 60115.
Mol Biol Res Commun. 2020 Dec;9(4):181-188. doi: 10.22099/mbrc.2020.38061.1530.
Newcastle Disease (ND) is a major viral disease in Indonesia. It is an RNA virus belongs to Paramyxovirinae. It is well known that RNA virus is easily to mutate. In some cases, this mutation could generate virulence alteration. It is noted that mutation of NDV which has avirulent amino acid sequence on the cleavage site, could mutate to be virulent Newcastle Disease Virus (NDV). It is needed to analyze the nucleotide and amino acid mutations and the effect of those to its virulence. The aim of this study was to analyze nucleotide and amino acid mutations of original isolated Lentogenic Newcastle Disease Virus (NDV). Samples were collected from cloacal swab of native chicken (Gallus gallus domesticus) suspected to be infected by Lentogenic NDV from live bird markets. They were inoculated into embryonated eggs, to isolate the virus. HA and HI assays were conducted to confirm that they were NDV. Positive samples were processed into serial passages in embryonated egg to observe their death time. Samples caused mortality of the embryonated eggs more than 90 hours post infection were suspected as Lentogenic NDV. They were processed to RT-PCR then sequenced. Lentogenic NDV confirmation was done by comparing amino acid at Fusion protein cleavage site of the samples to Lasota/JF950510. Nucleotide and amino acid mutations were analyzed. The result showed that some nucleotide mutations were capable to change sequences of amino acid but the virulence of the samples remained the same to the reference sequence.
新城疫（ND）是印度尼西亚一种主要的病毒性疾病。它是一种属于副粘病毒亚科的RNA病毒。众所周知，RNA病毒很容易发生变异。在某些情况下，这种变异可能导致毒力改变。值得注意的是，在裂解位点具有无毒氨基酸序列的新城疫病毒（NDV）可能会突变为强毒新城疫病毒（NDV）。需要分析核苷酸和氨基酸突变及其对毒力的影响。本研究的目的是分析原始分离的缓发型新城疫病毒（NDV）的核苷酸和氨基酸突变。样本取自活禽市场疑似感染缓发型NDV的本地鸡（家鸡）的泄殖腔拭子。将它们接种到鸡胚中以分离病毒。进行血凝（HA）和血凝抑制（HI）试验以确认它们是NDV。将阳性样本在鸡胚中进行连续传代以观察其死亡时间。感染后90小时以上导致鸡胚死亡的样本被怀疑为缓发型NDV。对它们进行逆转录聚合酶链反应（RT-PCR）然后测序。通过将样本融合蛋白裂解位点的氨基酸与LaSota/JF950510进行比较来确认缓发型NDV。分析核苷酸和氨基酸突变。结果表明，一些核苷酸突变能够改变氨基酸序列，但样本的毒力与参考序列相比保持不变。
