Volume 14, Issue 56 (9-2024)
NCMBJ 2024, 14(56): 95-106 |
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Shafiei N, Mahmoodzadeh Hosseini H, Amani J, Mirhosseini S A, Jafary H. Evaluation of binding affinity and sensitivity of two specific E. coli O157:H7 aptamers using the ELISA method. NCMBJ 2024; 14 (56) :95-106
URL: http://ncmbjpiau.ir/article-1-1706-en.html
URL: http://ncmbjpiau.ir/article-1-1706-en.html
Nafiseh Shafiei 
, Hamideh Mahmoodzadeh Hosseini , Jafar Amani , Seyed Ali Mirhosseini , Hanieh Jafary
, Hamideh Mahmoodzadeh Hosseini , Jafar Amani , Seyed Ali Mirhosseini , Hanieh Jafary
Applied Microbiology Research Center, Biomedicine Technologies institute, Baqiyatallah University of Medical Sciences, Tehran, Iran , hosseini361@yahoo.com
Abstract: (390 Views)
Aim and Background: Escherichia coli O157:H7 is one of the most dangerous foodborne pathogenic bacteria transmitted through food worldwide. Many detection techniques based on molecular and serological detection have limitations. In recent years, aptamers as small nucleic acid molecules have been utilized in many different applications and played an essential role as screening tools in various diagnostic systems. In this study, two aptamers were compared and evaluated for the detection of Escherichia coli cells.
Material and Methods: Specific primers were designed for each aptamer using the Oligo 7 software. The enzyme-linked immunosorbent assay (ELISA) was used to determine the detection limit and optimal concentration of the aptamers. The binding affinity of the aptamers to O157:H7 bacterial cells was investigated using a surface plasmon resonance instrument.
Results: The detection limits for E. Coli 73 and E. Coli 81 aptamers were determined to be 103 CFU/mL. The binding affinities of the E. Coli 73 and E. Coli 81 aptamers were 1.8137 E -9 and 5.1674E-8 mol/L, respectively.
Conclusion: Both aptamers showed good binding affinity to O157:H7 bacterial cells. However, the E. Coli 81 aptamer had higher sensitivity and specificity compared to the E. Coli 73 aptamer. Therefore, the E. Coli 81 aptamer could be a suitable candidate for the detection of O157:H7 bacterial cells in future applications.
Material and Methods: Specific primers were designed for each aptamer using the Oligo 7 software. The enzyme-linked immunosorbent assay (ELISA) was used to determine the detection limit and optimal concentration of the aptamers. The binding affinity of the aptamers to O157:H7 bacterial cells was investigated using a surface plasmon resonance instrument.
Results: The detection limits for E. Coli 73 and E. Coli 81 aptamers were determined to be 103 CFU/mL. The binding affinities of the E. Coli 73 and E. Coli 81 aptamers were 1.8137 E -9 and 5.1674E-8 mol/L, respectively.
Conclusion: Both aptamers showed good binding affinity to O157:H7 bacterial cells. However, the E. Coli 81 aptamer had higher sensitivity and specificity compared to the E. Coli 73 aptamer. Therefore, the E. Coli 81 aptamer could be a suitable candidate for the detection of O157:H7 bacterial cells in future applications.
Type of Study: Research Article |
Subject:
Cellular and molecular
Received: 2024/12/16 | Accepted: 2024/09/22 | Published: 2024/09/22
Received: 2024/12/16 | Accepted: 2024/09/22 | Published: 2024/09/22
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