tetano
Editor, Senior Moderator
J Med Virol
. 2025 Apr;97(4):e70327.
doi: 10.1002/jmv.70327. Whole-Genome Conservation Analysis for the Specific and Accurate Detection of Influenza A and B Viruses and Respiratory Syncytial Virus by Quadruplex RT-qPCR
Yun Zhang[SUP] 1 2 [/SUP], Chunhui Huo[SUP] 2 [/SUP], Jin Xiong[SUP] 3 [/SUP], Dongyan Xiong[SUP] 3 4 [/SUP], Ping He[SUP] 3 4 [/SUP], Zhanlong Wu[SUP] 3 4 [/SUP], Genqing Yang[SUP] 1 2 [/SUP], Junhua Li[SUP] 3 [/SUP], Hongping Wei[SUP] 3 4 [/SUP], Junping Yu[SUP] 2 3 4 [/SUP]
Affiliations
Influenza virus (Flu) and respiratory syncytial virus (RSV) are the primary pathogens responsible for acute respiratory infections. Both viruses are prone to mutations due to the seasonal epidemic, leading to an increasing rate of false-negative results. In this study, comprehensive meta-analyses of the genomes focusing on most conserved fragments have been performed for the four seasonal influenza viruses (two subtypes of Flu A: H1N1 and H3N2; two subtypes of Flu B: Yamagata and Victoria) and the two types of RSV: RSVA and RSVB), respectively. The most conserved sequences of 200 bp were identified as targets of the designed primer/probe sets for RT-qPCR were screened and optimized. Good sensitivities of the optimized primer/probe sets were obtained with the limits of detections of 2.95, 2.82, 1.57, 2.8, 1.19, and 2.12 copies/reaction for H1N1, H3N2, Yamagata, Victoria, RSVA and RSVB, respectively. Eventually, quadruplex qPCR using the four designed primer/probe sets can achieve simultaneous screening of the four viruses at a single tube. Furthermore, the assay's good performance in detecting target viruses from clinical throat swab samples demonstrated its potential for diagnosis of these viruses. The method, based on the identified conserved sequences and primer/probe sets, can effectively reduce false-negative results and rapidly respond to these viruses during respiratory disease outbreaks, or even before their widespread emergence, which aid in preventing outbreaks and guiding clinical treatment.
Keywords: conserved sequences; genome; influenza virus; respiratory syncytial virus.
. 2025 Apr;97(4):e70327.
doi: 10.1002/jmv.70327. Whole-Genome Conservation Analysis for the Specific and Accurate Detection of Influenza A and B Viruses and Respiratory Syncytial Virus by Quadruplex RT-qPCR
Yun Zhang[SUP] 1 2 [/SUP], Chunhui Huo[SUP] 2 [/SUP], Jin Xiong[SUP] 3 [/SUP], Dongyan Xiong[SUP] 3 4 [/SUP], Ping He[SUP] 3 4 [/SUP], Zhanlong Wu[SUP] 3 4 [/SUP], Genqing Yang[SUP] 1 2 [/SUP], Junhua Li[SUP] 3 [/SUP], Hongping Wei[SUP] 3 4 [/SUP], Junping Yu[SUP] 2 3 4 [/SUP]
Affiliations
- PMID: 40159799
- DOI: 10.1002/jmv.70327
Influenza virus (Flu) and respiratory syncytial virus (RSV) are the primary pathogens responsible for acute respiratory infections. Both viruses are prone to mutations due to the seasonal epidemic, leading to an increasing rate of false-negative results. In this study, comprehensive meta-analyses of the genomes focusing on most conserved fragments have been performed for the four seasonal influenza viruses (two subtypes of Flu A: H1N1 and H3N2; two subtypes of Flu B: Yamagata and Victoria) and the two types of RSV: RSVA and RSVB), respectively. The most conserved sequences of 200 bp were identified as targets of the designed primer/probe sets for RT-qPCR were screened and optimized. Good sensitivities of the optimized primer/probe sets were obtained with the limits of detections of 2.95, 2.82, 1.57, 2.8, 1.19, and 2.12 copies/reaction for H1N1, H3N2, Yamagata, Victoria, RSVA and RSVB, respectively. Eventually, quadruplex qPCR using the four designed primer/probe sets can achieve simultaneous screening of the four viruses at a single tube. Furthermore, the assay's good performance in detecting target viruses from clinical throat swab samples demonstrated its potential for diagnosis of these viruses. The method, based on the identified conserved sequences and primer/probe sets, can effectively reduce false-negative results and rapidly respond to these viruses during respiratory disease outbreaks, or even before their widespread emergence, which aid in preventing outbreaks and guiding clinical treatment.
Keywords: conserved sequences; genome; influenza virus; respiratory syncytial virus.