Detection of Non-PCR Amplified <em>S. enteritidis</em> Genomic DNA from Food Matrices Using a Gold-Nanoparticle DNA Biosensor: A Proof-of-Concept Study
Bacterial pathogens pose an increasing food safety and bioterrorism concern. Current DNA detection methods utilizing sensitive nanotechnology and biosensors have shown excellent detection, but require expensive and time-consuming polymerase chain reaction (PCR) to amplify DNA targets; thus, a faster...
| Main Authors: | , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
MDPI AG
2012-08-01
|
| Series: | Sensors |
| Subjects: | |
| Online Access: | http://www.mdpi.com/1424-8220/12/8/10487 |
| _version_ | 1798041961880354816 |
|---|---|
| author | Evangelyn C. Alocilja Michael C. Huarng Sylvia A. Vetrone |
| author_facet | Evangelyn C. Alocilja Michael C. Huarng Sylvia A. Vetrone |
| author_sort | Evangelyn C. Alocilja |
| collection | DOAJ |
| description | Bacterial pathogens pose an increasing food safety and bioterrorism concern. Current DNA detection methods utilizing sensitive nanotechnology and biosensors have shown excellent detection, but require expensive and time-consuming polymerase chain reaction (PCR) to amplify DNA targets; thus, a faster, more economical method is still essential. In this <em>proof-of-concept</em> study, we investigated the ability of a gold nanoparticle-DNA (AuNP-DNA) biosensor to detect non-PCR amplified genomic <em>Salmonella enterica </em>serovar Enteritidis (<em>S. enteritidis</em>) DNA, from pure or mixed bacterial culture and spiked liquid matrices. Non-PCR amplified DNA was hybridized into sandwich-like structures (magnetic nanoparticles/DNA/AuNPs) and analyzed through detection of gold voltammetric peaks using differential pulse voltammetry. Our preliminary data indicate that non-PCR amplified genomic DNA can be detected at a concentration as low as 100 ng/mL from bacterial cultures and spiked liquid matrices, similar to reported PCR amplified detection levels. These findings also suggest that AuNP-DNA biosensors are a first step towards a viable detection method of bacterial pathogens, in particular, for resource-limited settings, such as field-based or economically limited conditions. Future efforts will focus on further optimization of the DNA extraction method and AuNP-biosensors, to increase sensitivity at lower DNA target concentrations from food matrices comparable to PCR amplified DNA detection strategies. |
| first_indexed | 2024-04-11T22:28:52Z |
| format | Article |
| id | doaj.art-f11cfdbfe6b44c7ba2aed79afae166cb |
| institution | Directory Open Access Journal |
| issn | 1424-8220 |
| language | English |
| last_indexed | 2024-04-11T22:28:52Z |
| publishDate | 2012-08-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Sensors |
| spelling | doaj.art-f11cfdbfe6b44c7ba2aed79afae166cb2022-12-22T03:59:34ZengMDPI AGSensors1424-82202012-08-01128104871049910.3390/s120810487Detection of Non-PCR Amplified <em>S. enteritidis</em> Genomic DNA from Food Matrices Using a Gold-Nanoparticle DNA Biosensor: A Proof-of-Concept StudyEvangelyn C. AlociljaMichael C. HuarngSylvia A. VetroneBacterial pathogens pose an increasing food safety and bioterrorism concern. Current DNA detection methods utilizing sensitive nanotechnology and biosensors have shown excellent detection, but require expensive and time-consuming polymerase chain reaction (PCR) to amplify DNA targets; thus, a faster, more economical method is still essential. In this <em>proof-of-concept</em> study, we investigated the ability of a gold nanoparticle-DNA (AuNP-DNA) biosensor to detect non-PCR amplified genomic <em>Salmonella enterica </em>serovar Enteritidis (<em>S. enteritidis</em>) DNA, from pure or mixed bacterial culture and spiked liquid matrices. Non-PCR amplified DNA was hybridized into sandwich-like structures (magnetic nanoparticles/DNA/AuNPs) and analyzed through detection of gold voltammetric peaks using differential pulse voltammetry. Our preliminary data indicate that non-PCR amplified genomic DNA can be detected at a concentration as low as 100 ng/mL from bacterial cultures and spiked liquid matrices, similar to reported PCR amplified detection levels. These findings also suggest that AuNP-DNA biosensors are a first step towards a viable detection method of bacterial pathogens, in particular, for resource-limited settings, such as field-based or economically limited conditions. Future efforts will focus on further optimization of the DNA extraction method and AuNP-biosensors, to increase sensitivity at lower DNA target concentrations from food matrices comparable to PCR amplified DNA detection strategies.http://www.mdpi.com/1424-8220/12/8/10487biosensorgold-nanoparticlesDNA probegenomic DNAfood matrix |
| spellingShingle | Evangelyn C. Alocilja Michael C. Huarng Sylvia A. Vetrone Detection of Non-PCR Amplified <em>S. enteritidis</em> Genomic DNA from Food Matrices Using a Gold-Nanoparticle DNA Biosensor: A Proof-of-Concept Study Sensors biosensor gold-nanoparticles DNA probe genomic DNA food matrix |
| title | Detection of Non-PCR Amplified <em>S. enteritidis</em> Genomic DNA from Food Matrices Using a Gold-Nanoparticle DNA Biosensor: A Proof-of-Concept Study |
| title_full | Detection of Non-PCR Amplified <em>S. enteritidis</em> Genomic DNA from Food Matrices Using a Gold-Nanoparticle DNA Biosensor: A Proof-of-Concept Study |
| title_fullStr | Detection of Non-PCR Amplified <em>S. enteritidis</em> Genomic DNA from Food Matrices Using a Gold-Nanoparticle DNA Biosensor: A Proof-of-Concept Study |
| title_full_unstemmed | Detection of Non-PCR Amplified <em>S. enteritidis</em> Genomic DNA from Food Matrices Using a Gold-Nanoparticle DNA Biosensor: A Proof-of-Concept Study |
| title_short | Detection of Non-PCR Amplified <em>S. enteritidis</em> Genomic DNA from Food Matrices Using a Gold-Nanoparticle DNA Biosensor: A Proof-of-Concept Study |
| title_sort | detection of non pcr amplified lt em gt s enteritidis lt em gt genomic dna from food matrices using a gold nanoparticle dna biosensor a proof of concept study |
| topic | biosensor gold-nanoparticles DNA probe genomic DNA food matrix |
| url | http://www.mdpi.com/1424-8220/12/8/10487 |
| work_keys_str_mv | AT evangelyncalocilja detectionofnonpcramplifiedltemgtsenteritidisltemgtgenomicdnafromfoodmatricesusingagoldnanoparticlednabiosensoraproofofconceptstudy AT michaelchuarng detectionofnonpcramplifiedltemgtsenteritidisltemgtgenomicdnafromfoodmatricesusingagoldnanoparticlednabiosensoraproofofconceptstudy AT sylviaavetrone detectionofnonpcramplifiedltemgtsenteritidisltemgtgenomicdnafromfoodmatricesusingagoldnanoparticlednabiosensoraproofofconceptstudy |