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Abstract：
Equine herpesvirus myeloencephalopathy (EHM), a major problem for the equine industry in the United States, is caused by equine herpesvirus-1 (EHV-1). In addition, EHV-1 is associated with upper respiratory disease, abortion, and chorioretinal lesions in horses. Here we describe the development and evaluation of an inexpensive, user-friendly insulated isothermal PCR (iiPCR) method targeting open reading 30 (ORF30) to detect both neuropathogenic and non-neuropathogenic strains on the field-deployable POCKIT™ device for point-of-need detection of EHV-1. The analytical sensitivity of the EHV-1 iiPCR assay was 13 genome equivalents per reaction. The assay did not cross react with ten non-target equine viral pathogens. Performance of the EHV-1 iiPCR assay was compared to two previously described real-time PCR (qPCR) assays in two laboratories by using 104 archived clinical samples. All 53 qPCR-positive and 46 of the 51 qPCR-negative samples tested positive and negative, respectively, by the iiPCR. The agreement between the two assays was 95.19% (confidence interval 90.48-99.90%) with a kappa value of 0.90. In conclusion, the newly developed EHV-1 iiPCR assay is robust to provide specificity and sensitivity comparable to qPCR assays for the detection of EHV-1 nucleic acid in clinical specimens.
摘要：
马疱疹病毒性脊髓脑病（EHM），美国马业面临的一个主要问题，是由马疱疹病毒-1（EHV-1）引起的。此外,EHV-1与上呼吸道疾病有关,流产,和马的脉络膜视网膜病变.在这里，我们描述了廉价的开发和评估，针对开放读数30（ORF30）的用户友好型绝缘等温PCR（iiPCR）方法，用于在现场可部署的POCKIT™设备上检测神经致病性和非神经致病性菌株，用于需要点检测EHV-1。EHV-1iPCR测定的分析灵敏度为每个反应13个基因组当量。该测定不与十种非目标马病毒病原体交叉反应。通过使用104个存档的临床样品,将EHV-1iPCR测定的性能与两个实验室中的两个先前描述的实时PCR(qPCR)测定进行比较。所有53个qPCR阳性和51个qPCR阴性样本中的46个检测为阳性和阴性，分别,iPCR。两种测定之间的一致性为95.19%(置信区间90.48-99.90%),κ值为0.90。总之,新开发的EHV-1iPCR检测是稳健的,可提供与qPCR检测相当的特异性和灵敏度,用于检测临床标本中的EHV-1核酸.
