tetano
Editor, Senior Moderator
Anal Chem. 2015 Jun 30. [Epub ahead of print]
[h=1]Paper-based RNA extraction, in situ isothermal amplification, and lateral flow detection for low-cost, rapid diagnosis of Influenza A (H1N1) from clinical specimens.[/h] Rodriguez NM, Linnes JC, Fan A, Ellenson CK, Pollock NR, Klapperich CM.
[h=3]Abstract[/h] The 2009 Influenza A (H1N1) pandemic disproportionately affected the developing world and highlighted the key inadequacies of traditional diagnostic methods that make them unsuitable for use in resource-limited settings, from expensive equipment and infrastructure requirements to unacceptably long turnaround times. While rapid immunoassay diagnostic tests were much less costly and more context-appropriate, they suffered from drastically low sensitivities and high false negative rates. An accurate, sensitive and specific molecular diagnostic that is also rapid, low-cost, and independent of laboratory infrastructure is needed for effective point-of-care detection and epidemiological control in these developing regions. We developed a paper-based assay that allows for the extraction and purification of RNA directly from human clinical nasopharyngeal specimens through a polyethersulfone paper matrix, H1N1-specific in situ isothermal amplification directly within the same paper matrix, and immediate visual detection on lateral flow strips. The complete sample-to-answer assay can be performed at the point-of-care in just 45 minutes, without the need for expensive equipment or laboratory infrastructure, and has a clinically relevant viral load detection limit of 10^6 copies/mL, offering a 10-fold improvement over current rapid immunoassays.
PMID: 26125635 [PubMed - as supplied by publisher]
[h=1]Paper-based RNA extraction, in situ isothermal amplification, and lateral flow detection for low-cost, rapid diagnosis of Influenza A (H1N1) from clinical specimens.[/h] Rodriguez NM, Linnes JC, Fan A, Ellenson CK, Pollock NR, Klapperich CM.
[h=3]Abstract[/h] The 2009 Influenza A (H1N1) pandemic disproportionately affected the developing world and highlighted the key inadequacies of traditional diagnostic methods that make them unsuitable for use in resource-limited settings, from expensive equipment and infrastructure requirements to unacceptably long turnaround times. While rapid immunoassay diagnostic tests were much less costly and more context-appropriate, they suffered from drastically low sensitivities and high false negative rates. An accurate, sensitive and specific molecular diagnostic that is also rapid, low-cost, and independent of laboratory infrastructure is needed for effective point-of-care detection and epidemiological control in these developing regions. We developed a paper-based assay that allows for the extraction and purification of RNA directly from human clinical nasopharyngeal specimens through a polyethersulfone paper matrix, H1N1-specific in situ isothermal amplification directly within the same paper matrix, and immediate visual detection on lateral flow strips. The complete sample-to-answer assay can be performed at the point-of-care in just 45 minutes, without the need for expensive equipment or laboratory infrastructure, and has a clinically relevant viral load detection limit of 10^6 copies/mL, offering a 10-fold improvement over current rapid immunoassays.
PMID: 26125635 [PubMed - as supplied by publisher]