College of Animal Science and Technology, Jilin Agricultural University, Changchun, 130118, Jilin, China.
College of Chinese Medicine Materials, Jilin Agricultural University, Changchun, 130118, Jilin, China.
BMC Vet Res. 2024 May 7;20(1):180. doi: 10.1186/s12917-024-04025-8.
Infectious bovine rhinotracheitis (IBR), caused by Bovine alphaherpesvirus-1 (BoAHV-1), is an acute, highly contagious disease primarily characterized by respiratory tract lesions in infected cattle. Due to its severe pathological damage and extensive transmission, it results in significant economic losses in the cattle industry. Accurate detection of BoAHV-1 is of paramount importance. In this study, we developed a real-time fluorescent quantitative PCR detection method for detecting BoAHV-1 infections. Utilizing this method, we tested clinical samples and successfully identified and isolated a strain of BoAHV-1.1 from positive samples. Subsequently, we conducted a genetic evolution analysis on the isolate strain's gC, TK, gG, gD, and gE genes.
The study developed a real-time quantitative PCR detection method using SYBR Green II, achieving a detection limit of 7.8 × 10 DNA copies/μL. Specificity and repeatability analyses demonstrated no cross-reactivity with other related pathogens, highlighting excellent repeatability. Using this method, 15 out of 86 clinical nasal swab samples from cattle were found to be positive (17.44%), which was higher than the results obtained from conventional PCR detection (13.95%, 12/86). The homology analysis and phylogenetic tree analysis of the gC, TK, gG, gD, and gE genes of the isolated strain indicate that the JL5 strain shares high homology with the BoAHV-1.1 reference strains. Amino acid sequence analysis revealed that gC, gE, and gG each had two amino acid mutations, while the TK gene had one synonymous mutation and one H to Y mutation, with no amino acid mutations observed in the gD gene. Phylogenetic tree analysis indicated that the JL5 strain belongs to the BoAHV-1.1 genotype and is closely related to American strains such as C33, C14, and C28.
The established real-time fluorescent quantitative PCR detection method exhibits good repeatability, specificity, and sensitivity. Furthermore, genetic evolution analysis of the isolated BoAHV-1 JL-5 strain indicates that it belongs to the BoAHV-1.1 subtype. These findings provide a foundation and data for the detection, prevention, and control Infectious Bovine Rhinotracheitis.
传染性牛鼻气管炎（IBR）由牛α疱疹病毒 1 型（BoAHV-1）引起，是一种急性、高度传染性疾病，主要特征为感染牛的呼吸道病变。由于其严重的病理损伤和广泛传播，导致牛养殖业遭受重大经济损失。准确检测 BoAHV-1 至关重要。本研究开发了一种用于检测 BoAHV-1 感染的实时荧光定量 PCR 检测方法。利用该方法，我们对临床样本进行了检测，并成功从阳性样本中鉴定和分离出一株 BoAHV-1.1 株。随后，我们对分离株的 gC、TK、gG、gD 和 gE 基因进行了遗传进化分析。
本研究采用 SYBR Green II 建立了实时荧光定量 PCR 检测方法，检测限为 7.8×10 DNA 拷贝/μL。特异性和重复性分析表明，该方法与其他相关病原体无交叉反应，具有良好的重复性。使用该方法，从 86 份牛鼻拭子临床样本中检测到 15 份阳性（17.44%，15/86），高于常规 PCR 检测结果（13.95%，12/86）。分离株 gC、TK、gG、gD 和 gE 基因的同源性分析和系统进化树分析表明，JL5 株与 BoAHV-1.1 参考株高度同源。氨基酸序列分析表明，gC、gE 和 gG 各有 2 个氨基酸突变，TK 基因有 1 个同义突变和 1 个 H 到 Y 突变，gD 基因无氨基酸突变。系统进化树分析表明，JL5 株属于 BoAHV-1.1 基因型，与美国 C33、C14 和 C28 等株系密切相关。
本研究建立的实时荧光定量 PCR 检测方法具有良好的重复性、特异性和敏感性。此外，对分离的 BoAHV-1 JL-5 株的遗传进化分析表明，它属于 BoAHV-1.1 亚型。这些发现为传染性牛鼻气管炎的检测、预防和控制提供了基础和数据支持。
