Key result
The newly developed real-time RT-PCR assay detected 100% of human enterovirus-positive clinical specimens compared to a 13.3% detection rate by a commercial quantification kit.
Why the study?
Does the newly developed Taqman assay improve the detection and quantification of HRV viral load compared to a commercial kit in patients with acute respiratory infections?
Cross-Sectional (n=1,290)
No
Does the newly developed Taqman assay improve the detection and quantification of HRV viral load compared to a commercial kit in patients with acute respiratory infections?
Absolute Event Rate: 100% vs 13.3%
The newly developed Taqman assay significantly improves the detection and quantification of diverse HRV types compared to a standard commercial kit, revealing prolonged viral shedding and higher viral loads for HRV-C.
May enhance HRV detection in research; leaves open whether it warrants clinical adoption without prospective validation.
Human rhinovirus (HRV) is the major aetiology of respiratory tract infections. HRV viral load assays are available but limitations that affect accurate quantification exist. We developed a one-step Taqman assay using oligonucleotides designed based on a comprehensive list of global HRV sequences. The new oligonucleotides targeting the 5′-UTR region showed high PCR efficiency ( E = 99.6%, R 2 = 0.996), with quantifiable viral load as low as 2 viral copies/μl. Assay evaluation using an External Quality Assessment (EQA) panel yielded a detection rate of 90%. When tested on 315 human enterovirus-positive specimens comprising at least 84 genetically distinct HRV types/serotypes (determined by the VP4/VP2 gene phylogenetic analysis), the assay detected all HRV species and types, as well as other non-polio enteroviruses. A commercial quantification kit, which failed to detect any of the EQA specimens, produced a detection rate of 13.3% (42/315) among the clinical specimens. Using the improved assay, we showed that HRV sheds in the upper respiratory tract for more than a week following acute infection. We also showed that HRV-C had a significantly higher viral load at 2–7 days after the onset of symptoms ( p = 0.001). The availability of such assay is important to facilitate disease management, antiviral development, and infection control.
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Ng et al. (2016) conducted a cross-sectional in Acute upper respiratory tract infections (n=1,290). In-house real-time RT-PCR Taqman assay vs. Primerdesign genesig Human Rhinovirus All Subtypes kit was evaluated on Detection rate in human enterovirus-positive clinical specimens. The newly developed real-time RT-PCR assay detected 100% of human enterovirus-positive clinical specimens compared to a 13.3% detection rate by a commercial quantification kit.
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