title: A Portable, Pressure Driven, Room Temperature Nucleic Acid Extraction and Storage System for Point of Care Molecular Diagnostics.
abstract: Many new and exciting portable HIV viral load testing technologies are emerging for use in global medicine. While the potential to provide fast, isothermal, and quantitative molecular diagnostic information to clinicians in the field will soon be a reality, many of these technologies lack a robust front end for sample clean up and nucleic acid preparation. Such a technology would enable many different downstream molecular assays. Here, we present a portable system for centrifuge-free room temperature nucleic acid extraction from small volumes of whole blood (70 µL), using only thermally stable reagents compatible with storage and transport in low resource settings. Quantitative reverse transcriptase polymerase chain reaction (qRT-PCR) analysis of simulated samples demonstrate a lower limit of detection of 1000 copies/ml, with the ability to detect differences in viral load across four orders of magnitude. The system can also be used to store extracted RNA on detachable cartridges for up to one week at ambient temperature, and can be operated using only hand generated air pressure.
authors.affiliation: Department of Biomedical Engineering, Boston University, Boston, MA 02215.
authors.forename: Samantha
authors.initials: S
authors.lastname: Byrnes
authors.forename: Andy
authors.initials: A
authors.lastname: Fan
authors.forename: Jacob
authors.initials: J
authors.lastname: Trueb
authors.forename: Francis
authors.initials: F
authors.lastname: Jareczek
authors.forename: Mark
authors.initials: M
authors.lastname: Mazzochette
authors.forename: Andre
authors.initials: A
authors.lastname: Sharon
authors.forename: Alexis F
authors.initials: AF
authors.lastname: Sauer-Budge
authors.forename: Catherine M
authors.initials: CM
authors.lastname: Klapperich
keywords: DNA; HIV; PCR; RNA; blood; diagnostics; point of care; solid phase extraction
references.citation: J Acquir Immune Defic Syndr. 2008 Dec 15;49(5):477-84
references.pmid: 18989232
references.citation: Trends Biotechnol. 2011 May;29(5):240-50
references.pmid: 21377748
references.citation: J Infect Dis. 2010 Apr 15;201 Suppl 1:S16-26
references.pmid: 20225942
references.citation: Analyst. 2007 Dec;132(12):1193-9
references.pmid: 18318279
references.citation: BMC Infect Dis. 2008 Jul 04;8:89
references.pmid: 18601727
references.citation: AIDS. 2009 Nov 27;23(18):2459-66
references.pmid: 19741481
references.citation: Anal Chem. 2008 Nov 15;80(22):8453-60
references.pmid: 18855414
references.citation: Anal Chem. 2007 Aug 15;79(16):6135-42
references.pmid: 17622187
references.citation: Lab Chip. 2008 Dec;8(12):2121-7
references.pmid: 19023475
references.citation: Anal Chem. 2010 Jun 1;82(11):4344-56
references.pmid: 20443545
references.citation: Electrophoresis. 2002 Mar;23(5):727-33
references.pmid: 11891705
references.citation: J Clin Microbiol. 1998 Jan;36(1):258-60
references.pmid: 9431960
references.citation: J Clin Microbiol. 2009 Apr;47(4):1031-6
references.pmid: 19193847
references.citation: Anal Chem. 2010 Jul 1;82(13):5669-78
references.pmid: 20527816
references.citation: J Forensic Sci. 2006 Mar;51(2):266-73
references.pmid: 16566759
references.citation: Anal Chem. 2006 Oct 15;78(20):7222-8
references.pmid: 17037925
references.citation: Curr Opin HIV AIDS. 2010 Jan;5(1):1-5
references.pmid: 20046141
references.citation: Anal Chem. 2006 Aug 15;78(16):5704-10
references.pmid: 16906714
references.citation: Annu Rev Biomed Eng. 2008;10:107-44
references.pmid: 18358075
references.citation: AIDS. 2008 Oct 1;22(15):1971-7
references.pmid: 18784460
references.citation: Anal Chem. 2008 May 1;80(9):3483-91
references.pmid: 18355091
references.citation: Anal Chem. 2006 Mar 1;78(5):1673-81
references.pmid: 16503622
references.citation: Anal Chem. 2003 Apr 15;75(8):1880-6
references.pmid: 12713046
references.citation: J Clin Microbiol. 1990 Mar;28(3):495-503
references.pmid: 1691208
references.citation: Biotechnol Adv. 2010 Nov-Dec;28(6):770-81
references.pmid: 20600784
references.citation: PLoS Med. 2006 Oct;3(10):e417
references.pmid: 17032062
references.citation: Anal Chem. 2006 Feb 1;78(3):788-92
references.pmid: 16448052
references.citation: Trends Biochem Sci. 1996 Jun;21(6):224-5
references.pmid: 8744357
references.citation: Biosens Bioelectron. 2003 Oct 30;19(1):59-66
references.pmid: 14558999
references.citation: J Infect Dis. 2010 Apr 15;201 Suppl 1:S27-36
references.pmid: 20225943
references.citation: Clin Infect Dis. 2007 Jan 1;44(1):128-34
references.pmid: 17143828
references.citation: Anal Chem. 2009 Jul 1;81(13):5249-56
references.pmid: 19514712
references.citation: J Int AIDS Soc. 2010 Nov 05;13:43
references.pmid: 21054866
references.citation: Lab Chip. 2009 Oct 7;9(19):2811-7
references.pmid: 19967118
references.citation: Anal Biochem. 2000 Aug 1;283(2):175-91
references.pmid: 10906238
references.citation: Anal Biochem. 1987 Apr;162(1):156-9
references.pmid: 2440339