Shenzhen Key Laboratory of Pathogen and Immunity, State Key Discipline of Infectious Disease, Shenzhen Third People's Hospital, Second Hospital Affiliated to Southern University of Science and Technology, Shenzhen, China.
CAS Key Laboratory of Pathogenic Microbiology and Immunology, Collaborative Innovation Center for Diagnosis and Treatment of Infectious Disease, Center for Influenza Research and Early-warning (CASCIRE), Institute of Microbiology, Chinese Academy of Sciences, Beijing, China.
J Med Virol. 2020 Dec;92(12):3067-3072. doi: 10.1002/jmv.26133. Epub 2020 Jun 16.
Currently, two distinct lineages of influenza B virus (IBV), B/Victoria and B/Yamagata lineage, have been co-circulating in human beings. Assessment of the prevalent lineage is key for the recommendation of the seasonal influenza vaccine composition and the evaluation of its efficacy. In this study, a multiplex qRT-PCR assay for the discrimination of the IBV lineages was designed based on the genetic differences of the hemagglutinin genes between B/Yamagata and B/Victoria lineages. The assay was highly specific and able to discriminate the lineages of IBV without any non-specific reaction against other influenza A viruses. The detection limit of the assay was determined to be 10 genome-equivalent copies and 2.8 × 10 50% tissue culture infectious doses (TCID ) of live IBV per reaction. Moreover, our assay was able to discriminate the lineages of IBVs in clinical samples with 100% accuracy, when compared with pyrosequencing. Our results indicate that this assay may represent an update of the existing qRT-PCR assays and will be of great use for the rapid and accurate diagnosis and surveillance of the circulating IBVs.
目前，乙型流感病毒（IBV）有两个不同的谱系，B/Victoria 和 B/Yamagata 谱系，一直在人类中共同传播。评估流行谱系对于推荐季节性流感疫苗成分和评估其功效至关重要。在这项研究中，基于 B/Yamagata 和 B/Victoria 谱系之间血凝素基因的遗传差异，设计了一种用于区分 IBV 谱系的多重 qRT-PCR 检测方法。该检测方法具有高度特异性，能够区分 IBV 的谱系，而不会对其他甲型流感病毒产生任何非特异性反应。该检测方法的检测限为 10 个基因组等效拷贝和每个反应 2.8×10 50%组织培养感染剂量（TCID ）的活 IBV。此外，与焦磷酸测序相比，当用于临床样本时，我们的检测方法能够 100%准确地区分 IBV 的谱系。我们的结果表明，该检测方法可能代表现有 qRT-PCR 检测方法的更新，将非常有助于快速准确地诊断和监测循环中的 IBV。
