Hughes Holly R, Kenney Joan L, Russell Brandy J, Lambert Amy J
Arboviral Diseases Branch, Division of Vector-Borne Diseases, Centers for Disease Control and Prevention, Fort Collins, CO 80521, USA.
Pathogens. 2022 May 3;11(5):536. doi: 10.3390/pathogens11050536.
The neuroinvasive disease caused by Jamestown Canyon virus (JCV) infection is rare. However, increasing incidence and widespread occurrence of the infection make JCV a growing public health concern. Presently, clinical diagnosis is achieved through serological testing, and mosquito pool surveillance requires virus isolation and identification. A rapid molecular detection test, such as real-time RT-PCR, for diagnosis and surveillance of JCV has not been widely utilized. To enhance testing and surveillance, here, we describe the development and validation of a real-time RT-PCR test for the detection of JCV RNA. Three primer and probe sets were evaluated for analytical sensitivity and specificity. One probe set, JCV132FAM, was found to be the most sensitive test detecting 7.2 genomic equivalents/µL. While less sensitive, a second probe set JCV231cFAM was the most specific test with limited detection of Keystone virus at high RNA loads. Taken together, these data indicate both probe sets can be utilized for a primary sensitive screening assay and a secondary specific confirmatory assay. While both primer and probe sets detected high viral loads of Keystone virus, these assays did not detect any virus in the California encephalitis virus clade, including negative detection of the medically important La Crosse virus (LACV) and snowshoe hare virus (SSHV). The real-time RT-PCR assay described herein could be utilized in diagnosis and surveillance in regions with co-circulation of JCV and LACV or SSHV to inform public health action.
詹姆斯敦峡谷病毒（JCV）感染引起的神经侵袭性疾病较为罕见。然而，该感染发病率的上升和广泛传播使得JCV日益成为公共卫生关注的焦点。目前，临床诊断通过血清学检测实现，而蚊虫群落监测则需要病毒分离和鉴定。一种用于JCV诊断和监测的快速分子检测方法，如实时光RT-PCR，尚未得到广泛应用。为加强检测和监测，在此我们描述了一种用于检测JCV RNA的实时光RT-PCR检测方法的开发和验证。对三个引物和探针组进行了分析灵敏度和特异性评估。发现一个探针组JCV132FAM是最灵敏的检测方法，能检测到7.2个基因组当量/微升。第二个探针组JCV231cFAM虽然灵敏度较低，但在高RNA载量下对基斯通病毒的检测有限，是最特异的检测方法。综合来看，这些数据表明这两个探针组均可用于初步的灵敏筛选检测和二级特异性确证检测。虽然引物和探针组都检测到了高病毒载量的基斯通病毒，但这些检测方法在加利福尼亚脑炎病毒分支中未检测到任何病毒，包括对医学上重要的拉克罗斯病毒（LACV）和雪兔病毒（SSHV）的阴性检测。本文所述的实时光RT-PCR检测方法可用于JCV与LACV或SSHV共同流行地区的诊断和监测，以指导公共卫生行动。
