Department of Veterinary Microbiology, College of Veterinary and Animal Sciences, Udgir 413517, Maharashtra, India.
Department of Veterinary Microbiology, College of Veterinary and Animal Sciences, Parbhani 431402, Maharashtra, India.
Infect Genet Evol. 2019 Nov;75:103987. doi: 10.1016/j.meegid.2019.103987. Epub 2019 Aug 1.
Canine parvovirus (CPV) has emerged as an acute pathogen of young canine causing haemorrhagic enteritis and myocarditis. It is widely distributed and underreported in India. Therefore the study was conducted to type the CPV circulating in western Maharashtra. The faecal samples (n = 150) from clinically ill dogs showing diarrhoea and vomition were collected and subjected to haemagglutination (HA) with porcine RBC's. The DNA was extracted from the samples showing HA titres above 64 and subjected for amplification of VP gene fragment by PCR. The amplicons were subjected for restriction fragment length polymorphism (RFLP), sequencing and BEAST phylogenetic analysis. The results revealed 6% positivity by PCR. The RFLP results indicated single cleavage site for ApaLI and HinfI with an exception of two sites for HinfI. The nucleotide sequences showed nonfunctional nucleotide changes at different locations. The sequence analysis indicated that the nucleotide divergence within isolates under study was 0.00-0.42%, while the nucleotide homology was 99.58-100%. The most recent common ancestor was determined by molecular clock analysis using Bayesian methods. The sequence and phylogenetic analysis suggested the isolates as CPV-2a and KATN1 (KU866391, 2014) isolate from Tamilnadu, India as time to most recent common ancestor (TMRCA). The results revealed the circulating CPV in canines from western India as CPV2a genotype.
犬细小病毒 (CPV) 已成为导致幼犬急性出血性肠炎和心肌炎的病原体。它在印度广泛存在,但报告不足。因此,本研究旨在对印度西部流行的 CPV 进行分型。采集了 150 份来自出现腹泻和呕吐的临床患病犬的粪便样本,并用猪 RBC 进行血凝 (HA) 检测。从 HA 滴度高于 64 的样本中提取 DNA,进行 VP 基因片段的 PCR 扩增。对扩增子进行限制性片段长度多态性 (RFLP)、测序和 BEAST 系统发育分析。结果显示,PCR 检测阳性率为 6%。RFLP 结果表明,ApaLI 和 HinfI 只有一个单一的切割位点,但有两个位点除外。核苷酸序列显示在不同位置存在无功能的核苷酸变化。序列分析表明,研究中分离株的核苷酸差异在 0.00-0.42%之间,而核苷酸同源性为 99.58-100%。通过贝叶斯方法的分子钟分析确定了最近共同祖先的时间。序列和系统发育分析表明,这些分离株为 CPV-2a 基因型,与印度泰米尔纳德邦的 KATN1 (KU866391, 2014) 分离株具有相同的核苷酸序列。研究结果表明,来自印度西部的犬细小病毒为 CPV2a 基因型。