A previous sequence analysis of a US5 gene fragment of infectious laryngotracheitis virus (ILTV) performed in an Argentinian epidemiological study allowed to differentiate between wild and vaccine strains. This analysis also defined five ILTV haplotypes with specific variations at positions 461, 484, 832, 878 and 894 of the US5 gene. This characterization of viral strains may also be accomplished using the High-Resolution Melting Analysis (HRMA), which has been described as an effective, fast and sensitive method to detect mutations in PCR products. In the present study, an HRM protocol was developed with the aim of characterizing the circulating ILTV strains in Argentina. The specificity of this tool was confirmed in different DNA diluents, without interference from heterologous DNA or other cellular metabolites. Additionally, the salt concentration in the elution buffer used for DNA extraction did not alter the curve profiles. Higher concentrations of DNA (Ct ≅ 26.0) displayed well-defined curve profiles, whereas lower concentrations (Ct ≅ 32.5) exhibited more heterogeneous curves. The HRMA showed 97.49% concordance with the reference technique, i.e., sequencing. The HRM protocol has the capability to perform DNA amplification prior to its characterization. Thus, eventually this technique may be used simultaneously as a diagnostic tool. This advantage implies a significant reduction in the time and effort involved in sample processing.

之前在阿根廷流行病学研究中对传染性喉气管炎病毒 (ILTV) 的US5基因片段进行的序列分析可以区分野生毒株和疫苗毒株。该分析还定义了五种 ILTV 单倍型，在US5的第 461、484、832、878 和 894 位具有特定变异基因。病毒株的这种表征也可以使用高分辨率熔解分析 (HRMA) 来完成，它已被描述为检测 PCR 产物突变的有效、快速和灵敏的方法。在本研究中，开发了一个 HRM 协议，旨在表征阿根廷的循环 ILTV 菌株。该工具的特异性在不同的 DNA 稀释剂中得到证实，不受异源 DNA 或其他细胞代谢物的干扰。此外，用于 DNA 提取的洗脱缓冲液中的盐浓度不会改变曲线轮廓。较高浓度的 DNA (Ct  ≅  26.0) 显示出明确的曲线轮廓，而较低浓度 (Ct  ≅ 32.5）表现出更多的异质曲线。HRMA 与参考技术（即测序）的一致性为 97.49%。HRM 协议能够在其表征之前进行 DNA 扩增。因此，最终这种技术可以同时用作诊断工具。这一优势意味着显着减少样品处理所涉及的时间和精力。