Amaro, Moegiratul (2012) Immunoassay Based on Carbon Nanotubes/Gold Nanoparticles Nanocomposite for Salmonella enteric serovar Typhimurium Detection. Magister thesis, Universitas Brawijaya.
Abstract
Salmonella Enterica Serovar Typhimurium adalah salah satu bakteri patogen paling berbahaya yang mengarah pada wabah terkait dengan makanan. Oleh karena itu, uji yang efektif, presisi, cepat, selektif dan sensitif diperlukan. Untuk meningkatkan sensitivitas deteksi Typhimurium S. Enterica Serova, nanotube karbon multiwall (MWCNTS) dan nanopartikel emas (AUNPS) digabungkan dan digunakan untuk meningkatkan intensitas sinyal. Dalam penelitian ini, Immunoassay untuk S. Enterica Serovar Typhimurium dikembangkan menggunakan sistem deteksi scanometri. Nanokomposit MWCNT / AUNP dibuat oleh perakitan langsung AU 3+ untuk MWCNTS dan memungkinkan pertumbuhan AUNPS pada permukaan MWCNTS selama proses sintesis. Nanokomposit MWCNTS / AUNPS kemudian dilampirkan pada antibodi typhimurium anti-S. untuk digunakan sebagai molekul pendeteksian. Untuk sistem immunoassay, manik-manik magnetik fungsi streptavidin digunakan bersama dengan MWCNTS / AUNPS dan membran nitroselulosa digunakan sebagai dukungan yang solid. Beberapa parameter seperti pemblokiran reagen, sensitivitas dan selektivitas dioptimalkan. Memblokir MWCNTS / AUNPS dengan BSA 2% dan tanpa menghalangi perawatan menghasilkan intensitas sinyal yang tinggi. Sensitivitas tertinggi diperoleh dari reaksi tanpa memblokir pengobatan dan 2% BSA masing-masing pada 18 dan 42 CFU / ml, pada peningkatan 5 menit perak. Dengan menggunakan kompleks MBS / AB-biotin dan MWCNTS / AUNPS, ketika diterapkan pada sampel nyata memberikan batas deteksi rendah serendah 10.914 CFU / mL dalam susu biasa dan 134 CFU / mL dalam susu rendah lemak dengan pemblokiran BSA 2%. Immunoassay ini menggunakan kompleks MBS / AB-AB-Biotin dan MWCNTS menyediakan selektivitas tinggi terhadap S. Enterica Serovar Typhimurium karena tidak dapat mendeteksi panas yang terbunuh sel E.Coli dalam buffer, susu polos dan susu rendah lemak.
English Abstract
Salmonella enterica serovar Typhimurium is one of the most harmful pathogenic bacteria that leads to outbreak related with foods. Effective, precise, fast, selective and sensitive assay are therefore needed. To improve sensitivity of S. enterica serovar Typhimurium detection, multiwalled carbon nanotubes (MWCNTs) and gold nanoparticles (AuNPs) were combined and used to increase the signal intensity. In this study, immunoassay for S. enterica serovar Typhimurium was developed using a scanometric detection system. MWCNT/AuNP nanocomposite was fabricated by directly assembling of Au 3+ to MWCNTs and allowing the growth of AuNPs on the surface MWCNTs during synthesis process. The MWCNTs/AuNPs nanocomposite was then attached to anti- S. typhimurium antibody to be used as a detecting molecule. For the immunoassay system, streptavidin functionalized magnetic beads was used together with MWCNTs/AuNPs and nitrocellulose membrane were used as a solid support. Several parameters such as blocking reagents, sensitivity and selectivity were optimized. Blocking the MWCNTs/AuNPs with 2% BSA and without blocking treatment resulted in high signal intensity. The highest sensitivity was obtained from the reaction without blocking treatment and 2% BSA at 18 and 42 CFU/mL, respectively, at 5 minute silver enhancement. By using MBs/Ab-biotin and MWCNTs/AuNPs complex, when applied to real sample gave low limit of detection as low as 10,914 CFU/mL in plain milk and 134 CFU/mL in low fat milk with 2% BSA blocking. This immunoassay using MBs/Ab-biotin and MWCNTs/AuNPs complex provided high selectivity toward S. enterica serovar Typhimurium as it could not detect heat killed E.coli cells in buffer, plain milk and low fat milk.
| Item Type: | Thesis (Magister) |
|---|---|
| Identification Number: | TES/547/AMA/i/04101274 |
| Subjects: | 500 Natural sciences and mathematics > 547 Organic chemistry |
| Divisions: | S2/S3 > Magister Teknologi Hasil Pertanian, Fakultas Teknologi Pertanian |
| Depositing User: | Endro Setyobudi |
| Date Deposited: | 04 Sep 2012 13:59 |
| Last Modified: | 04 Sep 2012 13:59 |
| URI: | http://repository.ub.ac.id/id/eprint/157566 |
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