Abstract
Influenza virus and respiratory syncytial virus cause acute upper and lower respiratory tract infections, especially in children and the elderly. Early treatment for these infections is thought to be important, so simple and sensitive detection methods are needed for use at clinical sites. Therefore, in this study, real-time reverse transcription loop-mediated isothermal amplification assays with quenching primer for influenza virus and respiratory syncytial virus were developed. Evaluation of a total of 113 clinical specimens compared to real-time RT-PCR assays showed that the novel assays could distinguish between the types and subtypes of influenza virus and respiratory syncytial virus and had 100% diagnostic specificity. The diagnostic sensitivity of each assay exceeded 85.0% and the assays showed sufficient clinical accuracy. Furthermore, positive results could be obtained in around 15 min using the novel assays in cases with high concentrations of virus. The developed assays should be useful for identifying influenza virus and respiratory syncytial virus cases not only in experimental laboratories but also in hospital and quarantine laboratories.
Highlights
Influenza virus (IV) and respiratory syncytial virus (RSV) infections are common causes of acute upper and lower respiratory tract infections such as pneumonia and bronchiolitis and lead to high rates of hospitalization, especially in children and the elderly (Falsey and Walsh, 2000; Jain et al, 2015; Sugaya et al, 2000; Zhou et al, 2012)
A rapid, specific, and sensitive detection assay for IV and RSV using a novel rRT-loop-mediated isothermal amplification (LAMP) method was developed
The novel quenching primer (QPrimer)-based rRT-LAMP assays enabled the detection of each target gene at 25–250 copies/reaction at the lowest concentration (Table 4) and showed less sensitive than the reference reverse transcription PCR (rRT-PCR) assays which limit of detection were 6–9 copies/ reaction as determined in previous studies (Nakauchi et al, 2014,)
Summary
Influenza virus (IV) and respiratory syncytial virus (RSV) infections are common causes of acute upper and lower respiratory tract infections such as pneumonia and bronchiolitis and lead to high rates of hospitalization, especially in children and the elderly (Falsey and Walsh, 2000; Jain et al, 2015; Sugaya et al, 2000; Zhou et al, 2012). One newly developed test is the loop-mediated isothermal amplification (LAMP) method, a rapid and sensitive nucleic acid amplification method that is performed under isothermal conditions and requires less complicated equipment than PCR (Nagamine et al, 2002; Notomi et al, 2000). This method can yield results in less than 1 h and can be utilized for the detection of many kinds of viral genomes (Kurosaki et al, 2016; Shirato et al, 2014; Yamazaki et al, 2013).
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