45b829b964e33e70748eb63d944c5808 pmc_35049656.pdf 0af6aa92f9ed9b60e753b586a8d3164b245f237d pmc_35049656.pdf deac0c707f8175999a40de6350dc46584f806f04f5ab594ff6cc12470a4f0452 pmc_35049656.pdf Title: Tween 80 Improves the Acid-Fast Bacilli Quantification in the Magnetic Nanoparticle-Based Colorimetric Biosensing Assay (NCBA) Subject: Despite its reduced sensitivity, sputum smear microscopy (SSM) remains the main diagnostic test for detecting tuberculosis in many parts of the world. A new diagnostic technique, the magnetic nanoparticle-based colorimetric biosensing assay (NCBA) was optimized by evaluating different concentrations of glycan-functionalized magnetic nanoparticles (GMNP) and Tween 80 to improve the acid-fast bacilli (AFB) count. Comparative analysis was performed on 225 sputum smears: 30 with SSM, 107 with NCBA at different GMNP concentrations, and 88 with NCBA-Tween 80 at various concentrations and incubation times. AFB quantification was performed by adding the total number of AFB in all fields per smear and classified according to standard guidelines (scanty, 1+, 2+ and 3+). Smears by NCBA with low GMNP concentrations (1.5 mg/mL) showed higher AFB quantification compared to SSM. Cell enrichment of sputum samples by combining NCBA-GMNP, incubated with Tween 80 (5%) for three minutes, improved capture efficiency and increased AFB detection up to 445% over SSM. NCBA with Tween 80 offers the opportunity to improve TB diagnostics, mainly in paucibacillary cases. As this method provides biosafety with a simple and inexpensive methodology that obtains results in a short time, it might be considered as a point-of-care TB diagnostic method in regions where resources are limited. Keywords: tuberculosis; mycobacterium tuberculosis; tb detection; nanotechnology; sputum smear microscopy; tween 80; paucibacillary samples; increased sensitivity; diagnostic methods Author: Cristina Gordillo-Marroquín, Héctor J. Sánchez-Pérez, Anaximandro Gómez-Velasco, Miguel Martín, Karina Guillén-Navarro, Janeth Vázquez-Marcelín, Adriana Gómez-Bustamante, Letisia Jonapá-Gómez and Evangelyn C. Alocilja Creator: LaTeX with hyperref Producer: pdfTeX-1.40.21 CreationDate: Mon Jan 10 13:15:55 2022 CET ModDate: Tue Jan 11 02:04:12 2022 CET Tagged: no UserProperties: no Suspects: no Form: none JavaScript: no Pages: 18 Encrypted: no Page size: 595.276 x 841.89 pts (A4) Page rot: 0 File size: 5612037 bytes Optimized: no PDF version: 1.7 name type encoding emb sub uni object ID ------------------------------------ ----------------- ---------------- --- --- --- --------- HTHEIG+URWPalladioL-Bold Type 1 Custom yes yes no 10 0 HFPIYJ+URWPalladioL-Roma Type 1 Custom yes yes no 15 0 BDNEGC+URWPalladioL-Ital Type 1 Custom yes yes no 19 0 FFNIUC+CMSY10 Type 1 Builtin yes yes no 23 0 JYPLJJ+VnURWPalladioL-Bold Type 1 Custom yes yes no 27 0 MCBCCI+SourceSansRoman-Regular Type 1C Custom yes yes no 36 0 MCBCCM+SourceSansRoman.613wght Type 1C Custom yes yes no 40 0 MCBCCO+SourceSansRoman-Bold Type 1C Custom yes yes no 44 0 YHYESE+TimesNewRomanPS-BoldItalicMT Type 1C WinAnsi yes yes no 56 0 ZXDKIP+EURM10 Type 1 Builtin yes yes no 81 0 DUFXDP+VnURWPalladioL Type 1 Custom yes yes no 91 0 MFMLPH+PalatinoLinotype,Bold TrueType WinAnsi yes yes no 98 0 MFMLPJ+PalatinoLinotype TrueType WinAnsi yes yes no 101 0 MFMLPL+PalatinoLinotype,Italic TrueType WinAnsi yes yes no 104 0 BCDEEE+Calibri TrueType WinAnsi yes yes no 116 0 BCDGEE+PalatinoLinotype-Roman TrueType WinAnsi yes yes no 121 0 BCDHEE+Wingdings-Regular CID TrueType Identity-H yes yes yes 125 0 BCDIEE+PalatinoLinotype-Italic TrueType WinAnsi yes yes no 133 0 SymbolMT CID TrueType Identity-H yes no yes 137 0 CZKGAS+CMR10 Type 1 Builtin yes yes no 163 0 MFMLPH+PalatinoLinotype,Bold TrueType WinAnsi yes yes no 176 0 MFMLPI+PalatinoLinotype CID TrueType Identity-H yes yes yes 179 0 MFMLPJ+PalatinoLinotype TrueType WinAnsi yes yes no 185 0 MFMLPL+PalatinoLinotype,Italic TrueType WinAnsi yes yes no 188 0 MFMLPH+PalatinoLinotype,Bold TrueType WinAnsi yes yes no 198 0 MFMLPJ+PalatinoLinotype TrueType WinAnsi yes yes no 201 0 MFMLPL+PalatinoLinotype,Italic TrueType WinAnsi yes yes no 204 0 RDUGED+VnURWPalladioL-Italic Type 1 Custom yes yes no 227 0 Jhove (Rel. 1.22.1, 2019-04-17) Date: 2022-11-01 02:21:33 CET RepresentationInformation: pmc_35049656.pdf ReportingModule: PDF-hul, Rel. 1.12.1 (2019-04-17) LastModified: 2022-01-24 03:34:02 CET Size: 5612037 Format: PDF Version: 1.7 Status: Well-Formed and valid SignatureMatches: PDF-hul MIMEtype: application/pdf PDFMetadata: Objects: 513 FreeObjects: 1 IncrementalUpdates: 0 DocumentCatalog: PageLayout: SinglePage PageMode: UseNone Outlines: Item: Title: Introduction Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@4923ab24 Item: Title: Materials and Methods Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@44c8afef Children: Item: Title: Chemicals and Reagents Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@7b69c6ba Item: Title: Clinical Samples Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@46daef40 Item: Title: Processing of Sputum Samples Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@12f41634 Children: Item: Title: Sputum Smear Microscopy (SSM) Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@13c27452 Item: Title: Magnetic Nanoparticle-Based Colorimetric Biosensing Assay (NCBA) Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@262b2c86 Item: Title: Evaluation of the Tween 80 Effect in the Dispersion of GMNP-AFB Complexes Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@371a67ec Item: Title: AFB Quantification of Acid-Fast Bacilli on Smears Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@5ed828d Item: Title: Statistical Analysis Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@50d0686 Item: Title: Results Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@7a3d45bd Children: Item: Title: Analysis of AFB Detection in SSM and NCBA Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@1e7c7811 Item: Title: Low GMPN Concentrations Improve AFB Quantification Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@77ec78b9 Item: Title: Tween 80 at Low Concentration Improves the Quantification of AFB in NCBA Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@1a3869f4 Item: Title: Discussion Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@a38d7a3 Item: Title: Conclusions Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@77f99a05 Item: Title: References Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@63440df3 Info: Title: Tween 80 Improves the Acid-Fast Bacilli Quantification in the Magnetic Nanoparticle-Based Colorimetric Biosensing Assay (NCBA) Author: Cristina Gordillo-Marroquín, Héctor J. Sánchez-Pérez, Anaximandro Gómez-Velasco, Miguel Martín, Karina Guillén-Navarro, Janeth Vázquez-Marcelín, Adriana Gómez-Bustamante, Letisia Jonapá-Gómez and Evangelyn C. Alocilja Subject: Despite its reduced sensitivity, sputum smear microscopy (SSM) remains the main diagnostic test for detecting tuberculosis in many parts of the world. A new diagnostic technique, the magnetic nanoparticle-based colorimetric biosensing assay (NCBA) was optimized by evaluating different concentrations of glycan-functionalized magnetic nanoparticles (GMNP) and Tween 80 to improve the acid-fast bacilli (AFB) count. Comparative analysis was performed on 225 sputum smears: 30 with SSM, 107 with NCBA at different GMNP concentrations, and 88 with NCBA-Tween 80 at various concentrations and incubation times. AFB quantification was performed by adding the total number of AFB in all fields per smear and classified according to standard guidelines (scanty, 1+, 2+ and 3+). Smears by NCBA with low GMNP concentrations (1.5 mg/mL) showed higher AFB quantification compared to SSM. Cell enrichment of sputum samples by combining NCBA-GMNP, incubated with Tween 80 (5%) for three minutes, improved capture efficiency and increased AFB detection up to 445% over SSM. NCBA with Tween 80 offers the opportunity to improve TB diagnostics, mainly in paucibacillary cases. As this method provides biosafety with a simple and inexpensive methodology that obtains results in a short time, it might be considered as a point-of-care TB diagnostic method in regions where resources are limited. 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