Watanuki Sonoko, Bao Aronggaowa, Saitou Etsuko, Shoji Kazuyuki, Izawa Masaki, Okami Mitsuaki, Matsumoto Yasunobu, Aida Yoko
Laboratory of Global Infectious Diseases Control Science, Graduate School of Agricultural and Life Sciences, The University of Tokyo, 1-1-1 Yayoi, Bunkyo-ku, Tokyo 113-8657, Japan.
Hyogo Prefectural Awaji Meat Inspection Center, 49-18 Shitoorinagata, Minamiawaji 656-0152, Japan.
Pathogens. 2024 Dec 16;13(12):1111. doi: 10.3390/pathogens13121111.
The proviral load (PVL) of the bovine leukemia virus (BLV) is a useful index for estimating disease progression and transmission risk. Real-time quantitative PCR techniques are widely used for PVL quantification. We previously developed a dual-target detection method, the "Liquid Dual-CoCoMo assay", that uses the coordination of common motif (CoCoMo) degenerate primers. This method can detect two genes simultaneously using a FAM-labeled minor groove binder (MGB) probe for the BLV long terminal repeat (LTR) region and a VIC-labeled MGB probe for the gene. In this study, we evaluated the diagnostic and analytical performance of the Dual-CoCoMo assay targeting the LTR region by comparing its performance against the commercially available Takara multiplex assay targeting the region. The diagnostic sensitivity and specificity of the Liquid Dual-CoCoMo assay based on the diagnostic results of the ELISA or original Single-CoCoMo qPCR were higher than those of the Takara multiplex assay. Furthermore, using a BLV molecular clone, the analytical sensitivity of our assay was higher than that of the Takara multiplex assay. Our results provide the first evidence that the diagnostic and analytical performances of the Liquid Dual-CoCoMo assay are better than those of commercially available multiplex assays that target the region.
牛白血病病毒（BLV）的前病毒载量（PVL）是评估疾病进展和传播风险的有用指标。实时定量PCR技术广泛用于PVL定量。我们之前开发了一种双靶点检测方法，即“液体双CoCoMo检测法”，该方法使用共同基序（CoCoMo）简并引物的协同作用。该方法可以使用针对BLV长末端重复序列（LTR）区域的FAM标记的小沟结合剂（MGB）探针和针对 基因的VIC标记的MGB探针同时检测两个基因。在本研究中，我们通过将其性能与针对该区域的市售Takara多重检测法进行比较，评估了针对LTR区域的双CoCoMo检测法的诊断和分析性能。基于ELISA或原始单CoCoMo qPCR诊断结果的液体双CoCoMo检测法的诊断敏感性和特异性高于Takara多重检测法。此外，使用BLV分子克隆，我们检测法的分析敏感性高于Takara多重检测法。我们的结果首次证明，液体双CoCoMo检测法的诊断和分析性能优于针对该区域的市售多重检测法。
