Vagheshwari Dhaval H, Bhanderi Bharat B, Mathakiya Rafyuddin A, Jhala Mayurdhvaj K
Department of Veterinary Microbiology, College of Veterinary Science and Animal Husbandry, Anand Agricultural University, Anand, Gujarat 388001 India.
Virusdisease. 2017 Sep;28(3):320-327. doi: 10.1007/s13337-017-0387-3. Epub 2017 Jun 28.
The present study was undertaken with an aim of characterization of rabies virus (genus of the family under the order ) by sequencing of partial nucleoprotein (N) gene of rabies virus and phylogenetic analysis to know the genotype and lineage of rabies virus present in Gujarat state of India. A total of 32 samples (18 brain samples and 14 saliva samples) were aseptically collected from live and dead animals (viz. dog, buffalo, cow, goat, donkey and hyena) for rabies virus detection. Out of 32 samples, 24 samples were found positive by Reverse Transcriptase Polymerase Chain Reactions and from these 24 positive samples, 20 samples were selected for sequencing having good concentration of gene product. ClustalW alignment of nucleotide sequences and amino acid sequences of field rabies isolates revealed 95.20-100 and 97.95-100% similarity among themselves, respectively. Multiple sequence alignment of field rabies isolates and reference vaccine strains [Pasteur strain and Challenge Virus Strain (CVS)] indicated single nucleotide mutations at total 91 positions and amino acid mutations at total 17 different positions. Phylogenetic analysis of N gene sequences using our 20 field rabies isolates and 21 other reported isolates in Genbank resulted in 3 phylogenetic clusters. All the field rabies isolates showed same genetic lineage among themselves and with other earlier reported Indian rabies isolates placing them in Arctic like lineage of Genotype 1 Rabies virus. However, they were at genetic distance with reference Pasteur and CVS strains, which grouped in different phylogenetic cluster.
本研究旨在通过对狂犬病病毒部分核蛋白（N）基因进行测序以及系统发育分析，以鉴定狂犬病病毒（属于目 下的科 的属），从而了解印度古吉拉特邦存在的狂犬病病毒的基因型和谱系。为检测狂犬病病毒，共无菌采集了32份样本（18份脑样本和14份唾液样本），这些样本来自活体和死亡动物（即狗、水牛、奶牛、山羊、驴和鬣狗）。在32份样本中，通过逆转录聚合酶链反应发现24份样本呈阳性，从这24份阳性样本中，选择了20份基因产物浓度良好的样本进行测序。对现场狂犬病分离株的核苷酸序列和氨基酸序列进行ClustalW比对，结果显示它们之间的相似性分别为95.20 - 100%和97.95 - 100%。现场狂犬病分离株与参考疫苗株[巴斯德株和攻击病毒株（CVS）]的多序列比对表明，在总共91个位置存在单核苷酸突变，在总共17个不同位置存在氨基酸突变。使用我们的20个现场狂犬病分离株和Genbank中其他21个已报道的分离株对N基因序列进行系统发育分析，结果形成了3个系统发育簇。所有现场狂犬病分离株彼此之间以及与其他早期报道的印度狂犬病分离株显示出相同的遗传谱系，将它们归入基因型1狂犬病病毒的北极样谱系。然而，它们与参考巴斯德株和CVS株存在遗传距离，这些参考株归入不同的系统发育簇。
