Details
| Originalsprache | Englisch |
|---|---|
| Seiten (von - bis) | 923-930 |
| Seitenumfang | 8 |
| Fachzeitschrift | Engineering in life sciences |
| Jahrgang | 17 |
| Ausgabenummer | 8 |
| Publikationsstatus | Veröffentlicht - 6 Juli 2017 |
Abstract
Detection of food toxins with high sensitivity is very important and challenging. Ochratoxin A (OTA) is frequently present as food contaminant in contaminated grains and grain derivatives such as bread and beer. In this work, a target-induced dissociation (TID) based aptamer-assisted real-time PCR-based assay (apta-qPCR) is developed that features effective detection of OTA. Apta-qPCR effectively combines the capabilities of aptamer to be amplified, being a nucleotide sequence, with its specific interaction with the corresponding target molecule. Compared to commonly used fluorescence-based and colorimetric methods, the sensitivity of qPCR to detect a nucleotide sequence (aptamer) has ameliorated the sensitivity of the aptamer-based detection of OTA. Here, the OTA aptamer was immobilized on the magnetic beads coated with d(T)25 (dT beads). A sequence complementary to the OTA-binding portion of the aptamer was used as a linker between dT beads and the aptamer sequence. When OTA was added, the aptamer was released from the dT beads due to TID. The resulting assay was able to detect 0.009 ng/mL OTA with a wide dynamic range of 0.039–1000 ng/mL. Apta-qPCR can be easily transferred to other small molecules for highly sensitive detection using corresponding aptamers.
ASJC Scopus Sachgebiete
- Biochemie, Genetik und Molekularbiologie (insg.)
- Biotechnologie
- Umweltwissenschaften (insg.)
- Environmental engineering
- Chemische Verfahrenstechnik (insg.)
- Bioengineering
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in: Engineering in life sciences, Jahrgang 17, Nr. 8, 06.07.2017, S. 923-930.
Publikation: Beitrag in Fachzeitschrift › Artikel › Forschung › Peer-Review
}
TY - JOUR
T1 - Detection of ochratoxin A by aptamer-assisted real-time PCR-based assay (Apta-qPCR)
AU - Modh, Harshvardhan
AU - Scheper, Thomas
AU - Walter, Johanna Gabriela
N1 - Funding information: The German Research Foundation (DFG—SCHE 279/32-1) supported parts of this work. German Academic Exchange Service (DAAD) is acknowledged for the financial support to Harshvardhan Modh.
PY - 2017/7/6
Y1 - 2017/7/6
N2 - Detection of food toxins with high sensitivity is very important and challenging. Ochratoxin A (OTA) is frequently present as food contaminant in contaminated grains and grain derivatives such as bread and beer. In this work, a target-induced dissociation (TID) based aptamer-assisted real-time PCR-based assay (apta-qPCR) is developed that features effective detection of OTA. Apta-qPCR effectively combines the capabilities of aptamer to be amplified, being a nucleotide sequence, with its specific interaction with the corresponding target molecule. Compared to commonly used fluorescence-based and colorimetric methods, the sensitivity of qPCR to detect a nucleotide sequence (aptamer) has ameliorated the sensitivity of the aptamer-based detection of OTA. Here, the OTA aptamer was immobilized on the magnetic beads coated with d(T)25 (dT beads). A sequence complementary to the OTA-binding portion of the aptamer was used as a linker between dT beads and the aptamer sequence. When OTA was added, the aptamer was released from the dT beads due to TID. The resulting assay was able to detect 0.009 ng/mL OTA with a wide dynamic range of 0.039–1000 ng/mL. Apta-qPCR can be easily transferred to other small molecules for highly sensitive detection using corresponding aptamers.
AB - Detection of food toxins with high sensitivity is very important and challenging. Ochratoxin A (OTA) is frequently present as food contaminant in contaminated grains and grain derivatives such as bread and beer. In this work, a target-induced dissociation (TID) based aptamer-assisted real-time PCR-based assay (apta-qPCR) is developed that features effective detection of OTA. Apta-qPCR effectively combines the capabilities of aptamer to be amplified, being a nucleotide sequence, with its specific interaction with the corresponding target molecule. Compared to commonly used fluorescence-based and colorimetric methods, the sensitivity of qPCR to detect a nucleotide sequence (aptamer) has ameliorated the sensitivity of the aptamer-based detection of OTA. Here, the OTA aptamer was immobilized on the magnetic beads coated with d(T)25 (dT beads). A sequence complementary to the OTA-binding portion of the aptamer was used as a linker between dT beads and the aptamer sequence. When OTA was added, the aptamer was released from the dT beads due to TID. The resulting assay was able to detect 0.009 ng/mL OTA with a wide dynamic range of 0.039–1000 ng/mL. Apta-qPCR can be easily transferred to other small molecules for highly sensitive detection using corresponding aptamers.
KW - Apta-qPCR
KW - Aptamer
KW - Food toxin
KW - Ochratoxin A
KW - Target-induced dissociation
UR - http://www.scopus.com/inward/record.url?scp=85026736829&partnerID=8YFLogxK
U2 - 10.1002/elsc.201700048
DO - 10.1002/elsc.201700048
M3 - Article
AN - SCOPUS:85026736829
VL - 17
SP - 923
EP - 930
JO - Engineering in life sciences
JF - Engineering in life sciences
SN - 1618-0240
IS - 8
ER -