tetano
Editor, Senior Moderator
BMC Infect Dis. 2015 Feb 25;15(1):93. doi: 10.1186/s12879-015-0818-y.
[h=1]A multiplex PCR assay for the detection of five influenza viruses using a dual priming oligonucleotide system.[/h] Ma X[SUP]1[/SUP], Xu H[SUP]2[/SUP], Shi L[SUP]3[/SUP], Yang P[SUP]4[/SUP], Zhang L[SUP]5[/SUP], Sun X[SUP]6[/SUP], Zhen W[SUP]7[/SUP], Hu K[SUP]8[/SUP].
[h=3]Author information[/h]
[h=3]Abstract[/h] [h=4]BACKGROUND:[/h] A cost-effective, accurate and rapid simultaneous multiplex assay is required for testing and diagnoses of conventional and emerging viruses in clinical virology laboratories. We developed and optimized a dual priming oligonucleotide (DPO) multiplex PCR assay for detecting influenza viruses including seasonal H1N1, 2009 pandemic H1N1, H3N2, influenza B and H5N1.
[h=4]METHODS:[/h] The optimized multiplex DPO PCR was used to detect 233 clinical human samples. The results were compared to those obtained with RT-qPCR, conventional PCR and immunochromatographic assay.
[h=4]RESULTS:[/h] Specificity analysis revealed that the DPO PCR assay amplified each target virus without any cross-amplification. Statistical analysis demonstrated that the multiplex DPO-PCR sensitivity was higher than for the immunochromatographic assay and lower than for qPCR, while no significant difference was observed compared with conventional PCR, when detecting influenza A and B. Additional experiments using the same sample panel indicated no significant differences between the number of positive samples detected by multiplex DPO PCR and RT-qPCR when applying a Cq with a value lower than 30.
[h=4]CONCLUSIONS:[/h] The five-targeted simultaneous multiplex DPO PCR assay could be easily adopted into routine practice. This approach is cost effective with a short running time, low technical requirements for the detection of influenza virus and early diagnosis in clinical laboratories.
PMID: 25886516 [PubMed - in process] PMCID: PMC4344991
[h=1]A multiplex PCR assay for the detection of five influenza viruses using a dual priming oligonucleotide system.[/h] Ma X[SUP]1[/SUP], Xu H[SUP]2[/SUP], Shi L[SUP]3[/SUP], Yang P[SUP]4[/SUP], Zhang L[SUP]5[/SUP], Sun X[SUP]6[/SUP], Zhen W[SUP]7[/SUP], Hu K[SUP]8[/SUP].
[h=3]Author information[/h]
[h=3]Abstract[/h] [h=4]BACKGROUND:[/h] A cost-effective, accurate and rapid simultaneous multiplex assay is required for testing and diagnoses of conventional and emerging viruses in clinical virology laboratories. We developed and optimized a dual priming oligonucleotide (DPO) multiplex PCR assay for detecting influenza viruses including seasonal H1N1, 2009 pandemic H1N1, H3N2, influenza B and H5N1.
[h=4]METHODS:[/h] The optimized multiplex DPO PCR was used to detect 233 clinical human samples. The results were compared to those obtained with RT-qPCR, conventional PCR and immunochromatographic assay.
[h=4]RESULTS:[/h] Specificity analysis revealed that the DPO PCR assay amplified each target virus without any cross-amplification. Statistical analysis demonstrated that the multiplex DPO-PCR sensitivity was higher than for the immunochromatographic assay and lower than for qPCR, while no significant difference was observed compared with conventional PCR, when detecting influenza A and B. Additional experiments using the same sample panel indicated no significant differences between the number of positive samples detected by multiplex DPO PCR and RT-qPCR when applying a Cq with a value lower than 30.
[h=4]CONCLUSIONS:[/h] The five-targeted simultaneous multiplex DPO PCR assay could be easily adopted into routine practice. This approach is cost effective with a short running time, low technical requirements for the detection of influenza virus and early diagnosis in clinical laboratories.
PMID: 25886516 [PubMed - in process] PMCID: PMC4344991