Microfluidic leukocyte isolation for gene expression analysis in critically ill hospitalized patients

Aman Russom, Palaniappan Sethu, Daniel Irimia, Michael N. Mindrinos, Steve E. Calvano, Iris Garcia, Celeste Finnerty, Cynthia Tannahill, Amer Abouhamze, Julie Wilhelmy, M. Cecilia López, Henry V. Baker, David Herndon, Stephen F. Lowry, Ronald V. Maier, Ronald W. Davis, Lyle L. Moldawer, Ronald G. Tompkins, Mehmet Toner, Paul E. Bankey & 41 others Timothy R. Billiar, Bernard H. Brownstein, David G. Camp, George Casella, Irshad H. Chaudry, Mashkoor Choudhry, J. Perren Cobb, Asit De, Constance Elson, Bradley Freeman, Richard L. Gamelli, Nicole S. Gibran, Douglas L. Hayden, Brian G. Harbrecht, Jureta W. Horton, William Hubbard, Jeffrey Johnson, Matthew B. Klein, James A. Lederer, Tanya Logvinenko, John A. Mannick, Philip H. Mason, Grace P. McDonald-Smith, Bruce A. McKinley, Carol Miller-Graziano, Joseph P. Minei, Ernest E. Moore, Frederick A. Moore, Avery B. Nathens, Grant E. O'Keefe, Laurence G. Rahme, Daniel G. Remick, David A. Schoenfeld, Michael B. Shapiro, Martin Schwacha, Geoffrey M. Silver, Richard D. Smith, John Storey, H. Shaw Warren, Michael A. West, Wenzhong Xiao

Research output: Contribution to journalArticle

25 Citations (Scopus)

Abstract

BACKGROUND: Microarray technology is becoming a powerful tool for diagnostic, therapeutic, and prognostic applications. There is at present no consensus regarding the optimal technique to isolate nucleic acids from blood leukocyte populations for subsequent expression analyses. Current collection and processing techniques pose significant challenges in the clinical setting. Here, we report the clinical validation of a novel microfluidic leukocyte nucleic acid isolation technique for gene expression analysis from critically ill, hospitalized patients that can be readily used on small volumes of blood. METHODS: We processed whole blood from hospitalized patients after burn injury and severe blunt trauma according to the microfluidic and standard macroscale leukocyte isolation protocol. Side-by-side comparison of RNA quantity, quality, and genome-wide expression patterns was used to clinically validate the microfluidic technique. RESULTS: When the microfluidic protocol was used for processing, sufficient amounts of total RNA were obtained for genome-wide expression analysis from 0.5 mL whole blood. We found that the leukocyte expression patterns from samples processed using the 2 protocols were concordant, and there was less variability introduced as a result of harvesting method than there existed between individuals. CONCLUSIONS: The novel microfluidic approach achieves leukocyte isolation in <25 min, and the quality of nucleic acids and genome expression analysis is equivalent to or surpasses that obtained from macroscale approaches. Microfluidics can significantly improve the isolation of blood leukocytes for genomic analyses in the clinical setting.

Original languageEnglish (US)
Pages (from-to)891-900
Number of pages10
JournalClinical Chemistry
Volume54
Issue number5
DOIs
StatePublished - May 1 2008

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Microfluidics
Critical Illness
Gene expression
Leukocytes
Blood
Gene Expression
Nucleic Acids
Genes
Genome
RNA
Nonpenetrating Wounds
Microarrays
Processing
Blood Volume
Technology
Wounds and Injuries
Population

ASJC Scopus subject areas

  • Clinical Biochemistry

Cite this

Russom, A., Sethu, P., Irimia, D., Mindrinos, M. N., Calvano, S. E., Garcia, I., ... Xiao, W. (2008). Microfluidic leukocyte isolation for gene expression analysis in critically ill hospitalized patients. Clinical Chemistry, 54(5), 891-900. https://doi.org/10.1373/clinchem.2007.099150

Microfluidic leukocyte isolation for gene expression analysis in critically ill hospitalized patients. / Russom, Aman; Sethu, Palaniappan; Irimia, Daniel; Mindrinos, Michael N.; Calvano, Steve E.; Garcia, Iris; Finnerty, Celeste; Tannahill, Cynthia; Abouhamze, Amer; Wilhelmy, Julie; López, M. Cecilia; Baker, Henry V.; Herndon, David; Lowry, Stephen F.; Maier, Ronald V.; Davis, Ronald W.; Moldawer, Lyle L.; Tompkins, Ronald G.; Toner, Mehmet; Bankey, Paul E.; Billiar, Timothy R.; Brownstein, Bernard H.; Camp, David G.; Casella, George; Chaudry, Irshad H.; Choudhry, Mashkoor; Cobb, J. Perren; De, Asit; Elson, Constance; Freeman, Bradley; Gamelli, Richard L.; Gibran, Nicole S.; Hayden, Douglas L.; Harbrecht, Brian G.; Horton, Jureta W.; Hubbard, William; Johnson, Jeffrey; Klein, Matthew B.; Lederer, James A.; Logvinenko, Tanya; Mannick, John A.; Mason, Philip H.; McDonald-Smith, Grace P.; McKinley, Bruce A.; Miller-Graziano, Carol; Minei, Joseph P.; Moore, Ernest E.; Moore, Frederick A.; Nathens, Avery B.; O'Keefe, Grant E.; Rahme, Laurence G.; Remick, Daniel G.; Schoenfeld, David A.; Shapiro, Michael B.; Schwacha, Martin; Silver, Geoffrey M.; Smith, Richard D.; Storey, John; Warren, H. Shaw; West, Michael A.; Xiao, Wenzhong.

In: Clinical Chemistry, Vol. 54, No. 5, 01.05.2008, p. 891-900.

Research output: Contribution to journalArticle

Russom, A, Sethu, P, Irimia, D, Mindrinos, MN, Calvano, SE, Garcia, I, Finnerty, C, Tannahill, C, Abouhamze, A, Wilhelmy, J, López, MC, Baker, HV, Herndon, D, Lowry, SF, Maier, RV, Davis, RW, Moldawer, LL, Tompkins, RG, Toner, M, Bankey, PE, Billiar, TR, Brownstein, BH, Camp, DG, Casella, G, Chaudry, IH, Choudhry, M, Cobb, JP, De, A, Elson, C, Freeman, B, Gamelli, RL, Gibran, NS, Hayden, DL, Harbrecht, BG, Horton, JW, Hubbard, W, Johnson, J, Klein, MB, Lederer, JA, Logvinenko, T, Mannick, JA, Mason, PH, McDonald-Smith, GP, McKinley, BA, Miller-Graziano, C, Minei, JP, Moore, EE, Moore, FA, Nathens, AB, O'Keefe, GE, Rahme, LG, Remick, DG, Schoenfeld, DA, Shapiro, MB, Schwacha, M, Silver, GM, Smith, RD, Storey, J, Warren, HS, West, MA & Xiao, W 2008, 'Microfluidic leukocyte isolation for gene expression analysis in critically ill hospitalized patients', Clinical Chemistry, vol. 54, no. 5, pp. 891-900. https://doi.org/10.1373/clinchem.2007.099150
Russom, Aman ; Sethu, Palaniappan ; Irimia, Daniel ; Mindrinos, Michael N. ; Calvano, Steve E. ; Garcia, Iris ; Finnerty, Celeste ; Tannahill, Cynthia ; Abouhamze, Amer ; Wilhelmy, Julie ; López, M. Cecilia ; Baker, Henry V. ; Herndon, David ; Lowry, Stephen F. ; Maier, Ronald V. ; Davis, Ronald W. ; Moldawer, Lyle L. ; Tompkins, Ronald G. ; Toner, Mehmet ; Bankey, Paul E. ; Billiar, Timothy R. ; Brownstein, Bernard H. ; Camp, David G. ; Casella, George ; Chaudry, Irshad H. ; Choudhry, Mashkoor ; Cobb, J. Perren ; De, Asit ; Elson, Constance ; Freeman, Bradley ; Gamelli, Richard L. ; Gibran, Nicole S. ; Hayden, Douglas L. ; Harbrecht, Brian G. ; Horton, Jureta W. ; Hubbard, William ; Johnson, Jeffrey ; Klein, Matthew B. ; Lederer, James A. ; Logvinenko, Tanya ; Mannick, John A. ; Mason, Philip H. ; McDonald-Smith, Grace P. ; McKinley, Bruce A. ; Miller-Graziano, Carol ; Minei, Joseph P. ; Moore, Ernest E. ; Moore, Frederick A. ; Nathens, Avery B. ; O'Keefe, Grant E. ; Rahme, Laurence G. ; Remick, Daniel G. ; Schoenfeld, David A. ; Shapiro, Michael B. ; Schwacha, Martin ; Silver, Geoffrey M. ; Smith, Richard D. ; Storey, John ; Warren, H. Shaw ; West, Michael A. ; Xiao, Wenzhong. / Microfluidic leukocyte isolation for gene expression analysis in critically ill hospitalized patients. In: Clinical Chemistry. 2008 ; Vol. 54, No. 5. pp. 891-900.
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abstract = "BACKGROUND: Microarray technology is becoming a powerful tool for diagnostic, therapeutic, and prognostic applications. There is at present no consensus regarding the optimal technique to isolate nucleic acids from blood leukocyte populations for subsequent expression analyses. Current collection and processing techniques pose significant challenges in the clinical setting. Here, we report the clinical validation of a novel microfluidic leukocyte nucleic acid isolation technique for gene expression analysis from critically ill, hospitalized patients that can be readily used on small volumes of blood. METHODS: We processed whole blood from hospitalized patients after burn injury and severe blunt trauma according to the microfluidic and standard macroscale leukocyte isolation protocol. Side-by-side comparison of RNA quantity, quality, and genome-wide expression patterns was used to clinically validate the microfluidic technique. RESULTS: When the microfluidic protocol was used for processing, sufficient amounts of total RNA were obtained for genome-wide expression analysis from 0.5 mL whole blood. We found that the leukocyte expression patterns from samples processed using the 2 protocols were concordant, and there was less variability introduced as a result of harvesting method than there existed between individuals. CONCLUSIONS: The novel microfluidic approach achieves leukocyte isolation in <25 min, and the quality of nucleic acids and genome expression analysis is equivalent to or surpasses that obtained from macroscale approaches. Microfluidics can significantly improve the isolation of blood leukocytes for genomic analyses in the clinical setting.",
author = "Aman Russom and Palaniappan Sethu and Daniel Irimia and Mindrinos, {Michael N.} and Calvano, {Steve E.} and Iris Garcia and Celeste Finnerty and Cynthia Tannahill and Amer Abouhamze and Julie Wilhelmy and L{\'o}pez, {M. Cecilia} and Baker, {Henry V.} and David Herndon and Lowry, {Stephen F.} and Maier, {Ronald V.} and Davis, {Ronald W.} and Moldawer, {Lyle L.} and Tompkins, {Ronald G.} and Mehmet Toner and Bankey, {Paul E.} and Billiar, {Timothy R.} and Brownstein, {Bernard H.} and Camp, {David G.} and George Casella and Chaudry, {Irshad H.} and Mashkoor Choudhry and Cobb, {J. Perren} and Asit De and Constance Elson and Bradley Freeman and Gamelli, {Richard L.} and Gibran, {Nicole S.} and Hayden, {Douglas L.} and Harbrecht, {Brian G.} and Horton, {Jureta W.} and William Hubbard and Jeffrey Johnson and Klein, {Matthew B.} and Lederer, {James A.} and Tanya Logvinenko and Mannick, {John A.} and Mason, {Philip H.} and McDonald-Smith, {Grace P.} and McKinley, {Bruce A.} and Carol Miller-Graziano and Minei, {Joseph P.} and Moore, {Ernest E.} and Moore, {Frederick A.} and Nathens, {Avery B.} and O'Keefe, {Grant E.} and Rahme, {Laurence G.} and Remick, {Daniel G.} and Schoenfeld, {David A.} and Shapiro, {Michael B.} and Martin Schwacha and Silver, {Geoffrey M.} and Smith, {Richard D.} and John Storey and Warren, {H. Shaw} and West, {Michael A.} and Wenzhong Xiao",
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T1 - Microfluidic leukocyte isolation for gene expression analysis in critically ill hospitalized patients

AU - Russom, Aman

AU - Sethu, Palaniappan

AU - Irimia, Daniel

AU - Mindrinos, Michael N.

AU - Calvano, Steve E.

AU - Garcia, Iris

AU - Finnerty, Celeste

AU - Tannahill, Cynthia

AU - Abouhamze, Amer

AU - Wilhelmy, Julie

AU - López, M. Cecilia

AU - Baker, Henry V.

AU - Herndon, David

AU - Lowry, Stephen F.

AU - Maier, Ronald V.

AU - Davis, Ronald W.

AU - Moldawer, Lyle L.

AU - Tompkins, Ronald G.

AU - Toner, Mehmet

AU - Bankey, Paul E.

AU - Billiar, Timothy R.

AU - Brownstein, Bernard H.

AU - Camp, David G.

AU - Casella, George

AU - Chaudry, Irshad H.

AU - Choudhry, Mashkoor

AU - Cobb, J. Perren

AU - De, Asit

AU - Elson, Constance

AU - Freeman, Bradley

AU - Gamelli, Richard L.

AU - Gibran, Nicole S.

AU - Hayden, Douglas L.

AU - Harbrecht, Brian G.

AU - Horton, Jureta W.

AU - Hubbard, William

AU - Johnson, Jeffrey

AU - Klein, Matthew B.

AU - Lederer, James A.

AU - Logvinenko, Tanya

AU - Mannick, John A.

AU - Mason, Philip H.

AU - McDonald-Smith, Grace P.

AU - McKinley, Bruce A.

AU - Miller-Graziano, Carol

AU - Minei, Joseph P.

AU - Moore, Ernest E.

AU - Moore, Frederick A.

AU - Nathens, Avery B.

AU - O'Keefe, Grant E.

AU - Rahme, Laurence G.

AU - Remick, Daniel G.

AU - Schoenfeld, David A.

AU - Shapiro, Michael B.

AU - Schwacha, Martin

AU - Silver, Geoffrey M.

AU - Smith, Richard D.

AU - Storey, John

AU - Warren, H. Shaw

AU - West, Michael A.

AU - Xiao, Wenzhong

PY - 2008/5/1

Y1 - 2008/5/1

N2 - BACKGROUND: Microarray technology is becoming a powerful tool for diagnostic, therapeutic, and prognostic applications. There is at present no consensus regarding the optimal technique to isolate nucleic acids from blood leukocyte populations for subsequent expression analyses. Current collection and processing techniques pose significant challenges in the clinical setting. Here, we report the clinical validation of a novel microfluidic leukocyte nucleic acid isolation technique for gene expression analysis from critically ill, hospitalized patients that can be readily used on small volumes of blood. METHODS: We processed whole blood from hospitalized patients after burn injury and severe blunt trauma according to the microfluidic and standard macroscale leukocyte isolation protocol. Side-by-side comparison of RNA quantity, quality, and genome-wide expression patterns was used to clinically validate the microfluidic technique. RESULTS: When the microfluidic protocol was used for processing, sufficient amounts of total RNA were obtained for genome-wide expression analysis from 0.5 mL whole blood. We found that the leukocyte expression patterns from samples processed using the 2 protocols were concordant, and there was less variability introduced as a result of harvesting method than there existed between individuals. CONCLUSIONS: The novel microfluidic approach achieves leukocyte isolation in <25 min, and the quality of nucleic acids and genome expression analysis is equivalent to or surpasses that obtained from macroscale approaches. Microfluidics can significantly improve the isolation of blood leukocytes for genomic analyses in the clinical setting.

AB - BACKGROUND: Microarray technology is becoming a powerful tool for diagnostic, therapeutic, and prognostic applications. There is at present no consensus regarding the optimal technique to isolate nucleic acids from blood leukocyte populations for subsequent expression analyses. Current collection and processing techniques pose significant challenges in the clinical setting. Here, we report the clinical validation of a novel microfluidic leukocyte nucleic acid isolation technique for gene expression analysis from critically ill, hospitalized patients that can be readily used on small volumes of blood. METHODS: We processed whole blood from hospitalized patients after burn injury and severe blunt trauma according to the microfluidic and standard macroscale leukocyte isolation protocol. Side-by-side comparison of RNA quantity, quality, and genome-wide expression patterns was used to clinically validate the microfluidic technique. RESULTS: When the microfluidic protocol was used for processing, sufficient amounts of total RNA were obtained for genome-wide expression analysis from 0.5 mL whole blood. We found that the leukocyte expression patterns from samples processed using the 2 protocols were concordant, and there was less variability introduced as a result of harvesting method than there existed between individuals. CONCLUSIONS: The novel microfluidic approach achieves leukocyte isolation in <25 min, and the quality of nucleic acids and genome expression analysis is equivalent to or surpasses that obtained from macroscale approaches. Microfluidics can significantly improve the isolation of blood leukocytes for genomic analyses in the clinical setting.

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