Semiquantitative Nucleic Acid Test with Simultaneous Isotachophoretic Extraction and Amplification

Analytical Chemistry
Andrew T BenderJonathan D Posner

Abstract

Nucleic acid amplification tests (NAATs) provide high diagnostic accuracy for infectious diseases and quantitative results for monitoring viral infections. The majority of NAATs require complex equipment, cold chain dependent reagents, and skilled technicians to perform the tests. This largely confines NAATs to centralized laboratories and can significantly delay appropriate patient care. Low-cost, point-of-care (POC) NAATs are especially needed in low-resource settings to provide patients with diagnosis and treatment planning in a single visit to improve patient care. In this work, we present a rapid POC NAAT with integrated sample preparation and amplification using electrokinetics and paper substrates. We use simultaneous isotachophoresis (ITP) and recombinase polymerase amplification (RPA) to rapidly extract, amplify, and detect target nucleic acids from serum and whole blood in a paper-based format. We demonstrate simultaneous ITP and RPA can consistently detect 5 copies per reaction in buffer and 10 000 copies per milliliter of human serum with no intermediate user steps. We also show preliminary extraction and amplification of DNA from whole blood samples. Our test is rapid (results in less than 20 min) and made from low...Continue Reading

References

Mar 1, 1990·Journal of Clinical Microbiology·R BoomJ van der Noordaa
Jun 15, 1973·Annals of the New York Academy of Sciences·F M EveraertsT P Verheggen
Jun 1, 1996·Nature Medicine·M S SaagP A Volberding
Sep 26, 1997·Hepatology : Official Journal of the American Association for the Study of Liver Diseases·D R Gretch
Aug 17, 1999·Journal of Clinical Microbiology·A AbeM Kohara
May 26, 2001·BMJ : British Medical Journal·C P Price
Jan 10, 2002·Clinical Microbiology Reviews·Anthony Moody
May 23, 2002·Gastroenterology·Jean-Michel Pawlotsky
Jul 13, 2004·EMBO Reports·Myriam VincentHuimin Kong
Sep 6, 2005·Clinica Chimica Acta; International Journal of Clinical Chemistry·Elizabeth A Mothershed, Anne M Whitney
Nov 30, 2005·BioTechniques·Valentina N KondratovaAnatoly V Lichtenstein
Jun 8, 2006·PLoS Biology·Olaf PiepenburgNiall A Armes
Dec 21, 2006·Lab on a Chip·Curtis D ChinSamuel K Sia
Dec 27, 2007·The Canadian Journal of Infectious Diseases & Medical Microbiology = Journal Canadien Des Maladies Infectieuses Et De La Microbiologie Médicale·Mel KrajdenMartin Petric
Mar 25, 2008·Annual Review of Biomedical Engineering·Paul YagerJohn Gerdes
May 17, 2008·Biomedical Microdevices·Raymond Mariella
Sep 3, 2009·Journal of Food Protection·Byron Brehm-StecherMary Lou Tortorello
Oct 17, 2009·Analytical Chemistry·Alexandre PersatJuan G Santiago
Jul 31, 2010·Clinical Microbiology and Infection : the Official Publication of the European Society of Clinical Microbiology and Infectious Diseases·R W Peeling, D Mabey
Jan 6, 2011·Archives of Pathology & Laboratory Medicine·Juan P Olano, David H Walker
Jan 12, 2011·Journal of Virological Methods·Arman NabatiyanDavid M Kelso
Mar 8, 2011·Trends in Biotechnology·Angelika NiemzDavid S Boyle
Jul 8, 2011·Lab on a Chip·Changchun LiuHaim H Bau
Jul 20, 2011·International Journal of Infectious Diseases : IJID : Official Publication of the International Society for Infectious Diseases·LeeAnne M LuftDeirdre L Church
Jun 12, 2012·Journal of Clinical Virology : the Official Publication of the Pan American Society for Clinical Virology·Milena EulerManfred Weidmann
Nov 15, 2013·Clinical Infectious Diseases : an Official Publication of the Infectious Diseases Society of America·Angela M CaliendoUNKNOWN Infectious Diseases Society of America (IDSA)
Dec 18, 2013·The Lancet Infectious Diseases·Paul K DrainIngrid V Bassett
Jan 22, 2014·Journal of Chromatography. a·Anita RogacsJuan G Santiago
May 16, 2014·Analytical Chemistry·Babak Y MoghadamJonathan D Posner
May 31, 2014·Analytical Chemistry·Zachary Austin CrannellRebecca Richards-Kortum
Jul 18, 2014·Journal of Clinical Microbiology·Allyson V RitchieHelen H Lee
Sep 27, 2014·Lab on a Chip·Tally Rosenfeld, Moran Bercovici
Oct 14, 2014·RSC Advances·Jacqueline C LinnesCatherine M Klapperich
Jan 6, 2015·The Journal of Microbiology·Tsugunori NotomiHidetoshi Kanda
Jun 25, 2015·Analytical Chemistry·John T ConnellyGeorge M Whitesides
Jun 26, 2015·Expert Review of Molecular Diagnostics·Gyorgy Abel
Sep 5, 2015·Lab on a Chip·Xiang LiRichard M Crooks

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Citations

May 17, 2019·Clinical Microbiology Reviews·Paul K DrainNigel Garrett
Jul 2, 2020·Molecules : a Journal of Synthetic Chemistry and Natural Product Chemistry·Siraprapa BoobphahomKwanwoo Shin
Feb 2, 2019·Analytical and Bioanalytical Chemistry·Tianyu DongFeng Li
Jan 6, 2021·Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences·Benjamin P SullivanJonathan D Posner
May 26, 2020·The Journal of Molecular Diagnostics : JMD·Andrew T BenderJonathan D Posner
Dec 10, 2020·Science China. Chemistry·Yongxi ZhaoChunhai Fan
Jan 17, 2019·ACS Central Science·Dionysios C ChristodouleasParthena Chorti

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