Wildlife Epidemiology Lab, College of Veterinary Medicine, University of Illinois Urbana-Champaign, Urbana, IL 61802, USA.
Wildlife Epidemiology Lab, College of Veterinary Medicine, University of Illinois Urbana-Champaign, Urbana, IL 61802, USA; Veterinary Diagnostic Lab, College of Veterinary Medicine, University of Illinois Urbana-Champaign, Urbana, IL 61802, USA.
J Virol Methods. 2024 Jun;327:114941. doi: 10.1016/j.jviromet.2024.114941. Epub 2024 Apr 8.
Emerging infectious diseases are a threat that contributes to the decline of global chelonian species. Herpesviruses are among the most impactful pathogens described in chelonians and are frequently associated with a range of presentations across hosts with the potential for severe morbidity and mortality. Trachemys herpesvirus 1 (TrHV1) has been reported in red-eared and yellow-bellied sliders (Trachemys scripta elegans and Trachemys scripta scripta, respectively) but is largely understudied. Invasive red-eared sliders may serve as a reservoir for transmission to sympatric native species. This study aimed to develop a sensitive and specific quantitative real-time PCR (qPCR) assay for the detection of TrHV1 DNA to aid in the characterization of the epidemiology of this virus in aquatic turtles. Two TaqMan-MGB FAM-dye labeled primer-probe sets were designed and evaluated using plasmid dilutions. The higher performing assay was specific for TrHV1 DNA and had a linear dynamic range of 1.0 × 10 to 1.0 × 10 copies per reaction with an R of 0.999, slope of -3.386, and efficiency of 97.39%. The limit of detection was 10 copies per reaction, and there was no loss of reaction efficiency in the presence of TrHV1-negative chelonian oral-cloacal DNA. Overall, the Trachemys herpesvirus 1 assay meets established criteria for acceptable qPCR assays and will be a valuable tool in characterizing the epidemiology of Trachemys herpesvirus 1 in chelonians.
新兴传染病是导致全球龟鳖类物种减少的威胁之一。疱疹病毒是龟鳖类动物中最具影响力的病原体之一，常与宿主的多种表现形式相关联，具有严重的发病率和死亡率的潜在风险。已在红耳龟和黄腹滑龟（分别为 Trachemys scripta elegans 和 Trachemys scripta scripta）中报告了龟疱疹病毒 1（TrHV1），但对其研究还很不充分。入侵性的红耳龟可能是将病毒传播给同种的本地物种的储主。本研究旨在开发一种敏感和特异的定量实时 PCR（qPCR）检测方法，用于检测 TrHV1 DNA，以帮助研究该病毒在水龟中的流行病学特征。设计并评估了两个 TaqMan-MGB FAM 染料标记的引物-探针集，使用质粒稀释进行评估。表现更好的检测方法特异性地针对 TrHV1 DNA，具有 1.0×10 至 1.0×10 拷贝/反应的线性动态范围，R 值为 0.999，斜率为-3.386，效率为 97.39%。检测限为每个反应 10 个拷贝，并且在存在 TrHV1 阴性龟口腔-泄殖腔 DNA 时，反应效率没有损失。总体而言，龟疱疹病毒 1 检测方法符合可接受的 qPCR 检测方法的标准，将成为在龟鳖类动物中研究龟疱疹病毒 1 流行病学的有价值的工具。
