Bugi Ratno Budiarto1, Apon Zaenal Mustopa2, Ratih Asmana Ningrum1, Nurul Amilia3,4, Endang Saepudin3. 1. Research Center for Genetic Engineering, National Research and Innovation Agency (BRIN), Raya Bogor Street KM.46, Cibinong, Bogor, West Java, 16911, Indonesia. 2. Research Center for Genetic Engineering, National Research and Innovation Agency (BRIN), Raya Bogor Street KM.46, Cibinong, Bogor, West Java, 16911, Indonesia. azmustopa@yahoo.com. 3. Department of Chemistry, Faculty of Mathematics and Natural Science, Universitas Indonesia, Depok, 16424, Indonesia. 4. Department of Chemistry, Faculty of Science and Technology, UIN Syarif Hidayatullah Jakarta, South Tangerang, 15412, Indonesia.
Abstract
BACKGROUND: Diarrhea is a major cause of severe gastrointestinal illness in the infant especially in many developing countries. Although this molecular technique has been accepted as standard technique to detect Diarrhea-causing EPEC, the practical aspect of this technique for in-site rapid screening purposes is still facing a major challenge. In this study, we characterized EPEC specific aptamers and applied it as an AuNP-based aptasensor for point of care (POC) diagnosis purpose. METHODS: As many as six selected DNA aptamers was screened using target bacteria and the bound aptamer was measured by qPCR technique. Moreover, Kd value for each optimal bound aptamer was measured by using the same technique. Colorimetry assay was applied to test specificity and LOD of AuNP-based aptasensor. RESULTS: Two DNA aptamers have been successfully obtained to detect Enteropathogenic Escherichia coli K.1.1. DNA aptamer S8-7 exhibited constant dissociation (Kd) value of 17.08 nM, while DNA aptamer S10-5 exhibited Kd value of 34.14 nM. AuNP-based aptasensor showed high selectivity and specificity for EPEC K.1.1 with a limit of detection (LOD) value of 105 CFU/mL. Truncation study on DNA aptamer S8-7 showed that elimination of primer binding sequence only slightly increased both performance of detection and LOD value of AuNP-based aptasensor. CONCLUSION: Further study is necessary to improve AuNP-aptasensor performance such as through mutagenesis approach on targeted DNA aptamers before AuNP-based aptasensor can be applied as a biosensor in point of care (POC) diagnosis.
BACKGROUND: Diarrhea is a major cause of severe gastrointestinal illness in the infant especially in many developing countries. Although this molecular technique has been accepted as standard technique to detect Diarrhea-causing EPEC, the practical aspect of this technique for in-site rapid screening purposes is still facing a major challenge. In this study, we characterized EPEC specific aptamers and applied it as an AuNP-based aptasensor for point of care (POC) diagnosis purpose. METHODS: As many as six selected DNA aptamers was screened using target bacteria and the bound aptamer was measured by qPCR technique. Moreover, Kd value for each optimal bound aptamer was measured by using the same technique. Colorimetry assay was applied to test specificity and LOD of AuNP-based aptasensor. RESULTS: Two DNA aptamers have been successfully obtained to detect Enteropathogenic Escherichia coli K.1.1. DNA aptamer S8-7 exhibited constant dissociation (Kd) value of 17.08 nM, while DNA aptamer S10-5 exhibited Kd value of 34.14 nM. AuNP-based aptasensor showed high selectivity and specificity for EPEC K.1.1 with a limit of detection (LOD) value of 105 CFU/mL. Truncation study on DNA aptamer S8-7 showed that elimination of primer binding sequence only slightly increased both performance of detection and LOD value of AuNP-based aptasensor. CONCLUSION: Further study is necessary to improve AuNP-aptasensor performance such as through mutagenesis approach on targeted DNA aptamers before AuNP-based aptasensor can be applied as a biosensor in point of care (POC) diagnosis.
Authors: Regiane C B Dias; Bruna C Dos Santos; Luis F Dos Santos; Melissa A Vieira; Ricardo S Yamatogi; Alessandro L Mondelli; Terue Sadatsune; José M Sforcin; Tânia A T Gomes; Rodrigo T Hernandes Journal: APMIS Date: 2016-01-11 Impact factor: 3.205
Authors: Teresa Estrada-Garcia; Catalina Lopez-Saucedo; Rocio Thompson-Bonilla; Maricela Abonce; Daniel Lopez-Hernandez; Jose Ignacio Santos; Jorge L Rosado; Herbert L DuPont; Kurt Z Long Journal: J Clin Microbiol Date: 2008-11-19 Impact factor: 5.948
Authors: Aamer Imdad; Monique A Foster; Junaid Iqbal; Christopher Fonnesbeck; Daniel C Payne; Chengxian Zhang; James D Chappell; Natasha Halasa; Oscar G Gómez-Duarte Journal: Pediatr Infect Dis J Date: 2018-06 Impact factor: 2.129