ISSN: 1859 - 2872
To establish a realtime polychain reaction-based assay for the detection of Human metapneumovirus
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Abstract
Objective: To validate a real-time RT-PCR assay for detection of Human metapneumovirus. Subject and method: This real-time PCR-based method was employed for the amplification of target genes, utilizing relevant negative controls, and a Human metapneumovirus plasmid. Detection limit and sensitivity were determined through a range of spiking assays covering a concentration range from 100 to 105 copies/µL. Primers and probe were designed using the hMPV sequences available in the NCBI database. A limited clinical samples were also used as input for the newly established assay to compare its performance with an IVD marked kit. Result: The limit of detection (LOD) of our in-house assay is 1 copy hMPV derived cDNA per microliter (µL), the assay showed a high level of specificity at 100% and a sensitivity of 95%. made it equal performance to that of Seegene's kit. Conclusion: We have successfully established a highly sensitive and specific real-time PCR assay for the detection of Human metapneumovirus using a positive standard plasmid template.
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References
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