A newly developed real‐time PCR assay for discriminating influenza B virus Yamagata and Victoria lineages

• Published in: Journal of medical virology Vol. 92; no. 12; pp. 3067 - 3072
• Main Authors: Wei, Jinli, Huang, Huaxin, Qin, Hucheng, Li, Shanqin, Xing, Li, Li, Yanjie, Cao, Mengli, Wang, Yanrong, Bi, Yuhai, Liu, Yingxia, Yang, Yang
• Format: Journal Article
• Language: English
• Published: United States Wiley Subscription Services, Inc 01.12.2020
• Subjects: Assaying; B/Victoria and B/Yamagata lineages; Diagnosis; discrimination; Equivalence; Genomes; Hemagglutinin Glycoproteins, Influenza Virus - genetics; Hemagglutinins; Humans; Influenza; Influenza A; Influenza B; influenza B virus; Influenza B virus - classification; Influenza B virus - genetics; Influenza B virus - isolation & purification; Influenza, Human - diagnosis; Influenza, Human - virology; Multiplex Polymerase Chain Reaction - methods; Multiplexing; Real-Time Polymerase Chain Reaction - methods; real‐time PCR; Sensitivity and Specificity; Tissue culture; Vaccines; Virology; Viruses; discrimination; real-time PCR; B/Victoria and B/Yamagata lineages; influenza B virus
• ISSN: 0146-6615; 1096-9071; 1096-9071
• DOI: 10.1002/jmv.26133

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 × 10‐2 50% tissue culture infectious doses (TCID50) 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. Highlights A multiplex qRT‐PCR assay was developed to discriminate the lineages of IBV with high sensitivity and broad coverage. The detection limit of the assay was determined to be 10 genome‐equivalent copies and 2.8 × 10‐2 50% tissue culture infectious doses (TCID50) of live IBV. This assaywill be of great use for the rapid and accurate diagnosis andsurveillance of the circulating IBVs.