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. 2025 May 15;10(5):3317–3339. doi: 10.1021/acssensors.5c00668

2. VF-μPADs for in Vitro Diagnostics.

Target analyte(s) Fabrication method Used materials Sensing modality Specimen (volume, μL) Detection limit Ref.
Small molecules: Iron Stacking Cellulose membrane, glass fiber, and polysulfone Colorimetric Whole blood (50 μL) Spiked sample:0.0004.AU μg–1 dL–1
Small molecules: Glucose, BSA Stacking Cellulose membrane Colorimetric PBS (80 μL) Buffer-based: 0.7 mM (glucose), 18 mM (BSA)
Small molecules: Glucose, Lactate Stacking Cellulose membrane Colorimetric PBS (15 μL) Buffer-based: 0.3 mM (glucose), 0.02 mM (lactate)
Small molecules: Methyl parathion Origami/folding Cellulose membrane Potentiometric 10 μL Spiked sample: 0.06 nM
Small molecules: Cyanide ion Origami/folding Cellulose membrane Colorimetric Borate buffer (2 μL) Buffer-based: 0.4 μmol/L
Small molecules: Mercury, Glucose Origami/folding Cellulose membrane Electrochemical Tap water (mercury, 10 μL), Sweat (glucose, 10 μL) NA
Small molecules: Glucose, Cholesterol, Albumin, ALP, Creatinine, AST, ALT, Urea Nitrogen 3D printing Cellulose membrane Colorimetric NA Buffer-based: 0.3 mmol/L (glucose), 0.3 mmol/L (cholesterol), 0.1 g/dL (albumin), 10 U/L (ALP), 40 μmol/L (creatinine), 10 U/L (AST), 50 U/L (ALT), 0.04 mmol/L (Urea Nitrogen)
Small molecules: Glucose 3D printing Cellulose membrane, and Polysulfone Colorimetric Whole blood (100 μL) Spiked sample: 0.3 mM
Small molecules: Dopamine 3D printing Cellulose membrane Fluorescence Tris-buffer (59 μL) NA
Small molecules: Glucose, Albumin 3D printing Cellulose membrane Colorimetric Artificial urine (5 μL) Buffer-based: 0.8 mM (glucose), 3.5 μM (albumin)
Protein-noninfectious disease: Myoglobin, creatine kinase-MB (CK-MB), and heart-type fatty acid binding protein (h-FABP) Stacking NC membrane, Polysulfone, and cotton fiber Fluorescence Serum (50 μL) Spiked sample: 0.52 ng/mL (Myoglobin), 0.3 ng/mL (CK-MB), 0.49 ng/mL (h-FABP)
Protein-noninfectious disease: Cardiac troponin I (cTnI) Stacking NC membrane, Polysulfone, and cotton fiber Colorimetric Serum (50 μL) Spiked sample: 0.2 pg/mL
Protein-noninfectious disease: Cardiac troponin I (cTnI) Stacking NC membrane, Polysulfone, and cotton fiber Chemiluminescence Serum (50 μL) Spiked sample: 0.16 pg/mL
Protein-noninfectious disease: Alkaline phosphatase, β-d-galactosidase Stacking Cellulose membrane Colorimetric Serum Spiked sample: 2.4 pM (alkaline phosphatase)
Protein-noninfectious disease: Blood alanine aminotransferase (ALT) Stacking Plasma separation membrane Colorimetric Whole blood (40 μL) Spiked sample: 53 U/L
Protein-noninfectious disease: C-reactive protein (CRP) Stacking Cellulose membrane, NC membrane Colorimetric Whole blood (100 μL) Spiked sample: 40 ng/mL
Protein-noninfectious disease: Ammonia, Carbon dioxide, Urea Origami/folding Cellulose membrane Colorimetric Saliva/plasma (2 μL) Spiked sample: 0.03 mg/dL (ammonia), 0.06 mg/dL (carbon dioxide), 0.18 mg/dL (urea)
Protein-noninfectious disease: Human serum albumin (HSA), Human immunoglobulin G (HIgG) Origami/folding Cellulose membrane Electrochemical Priming buffer (20 μL) Buffer-based: 1.5 pM
Protein-infectious disease: IgM/IgG antibodies for Lyme disease Stacking NC membrane, Polysulfone, and cotton fiber Colorimetric Serum (50 μL) Buffer-based:209.6 ng/mL (anti-OspC), 162.2 ng/mL (anti-BmpA), 1.05 μg/mL (anti-P41)
Protein-infectious disease: IgM/IgG antibodies for SARS-CoV-2 Stacking NC membrane, Polysulfone, and cotton fiber Colorimetric Serum (40 μL; 20 μL each for IgM and IgG tests) Undetermined
Protein-infectious disease: IgM/IgG antibodies for Lyme disease Stacking NC membrane, Polysulfone, and cotton fiber Colorimetric Serum (20 μL) Undetermined
Protein-infectious disease: H1N1 virus Stacking Cellulose membrane, Polytetrafluorethylene membrane Electrochemical, Colorimetric Saliva (40 μL) Spike sample: 4.7 PFU/mL by electrochemical method, 2.27 PFU/mL by colorimetric method
Protein-infectious disease: Human norovirus Stacking Cellulose/polyester paper, NC membrane Colorimetric PBS (110 μL) Buffer-based:  9.5 × 104 copies/mL
Protein-infectious disease: Protein A for S. aureus Origami/folding Cellulose membrane Colorimetric Real sample: Synovial fluid (3 μL) NA
Protein-infectious disease: Dengue virus serotypes 3D printing Cellulose membrane Colorimetric 45 μL Buffer-based: 23.28 nM (DENV-1), 5.23 nM (DENV-2), 38.17 nM (DENV-3), and 29.64 nM (DENV-4)
Nucleic acid: Hepatitis B Virus DNA Origami/folding Cellulose membrane Electrochemical Trailing electrolyte solution (50 μL) NA
Nucleic acid: SARS-CoV-2 RNA and its variants Origami/folding Cellulose membrane Colorimetric Molecular biology grade H2O (5 μL) Buffer-based: 400 copies/μL (SARS-CoV-2 RNA)
Nucleic acid: E. coli O157:H7, Salmonella Origami/folding Cellulose membrane Colorimetric 25 μL (bacterial solution), 15 μL (LAMP reaction) Buffer-based: 25 CFU