Detection of ochratoxin A using molecular beacons and real-time PCR thermal cycler

Toxins
Simona Marianna SanzaniAntonio Ippolito

Abstract

We developed a simple and cheap assay for quantitatively detecting ochratoxin A (OTA) in wine. A DNA aptamer available in literature was used as recognition probe in its molecular beacon form, i.e., with a fluorescence-quenching pair at the stem ends. Our aptabeacon could adopt a conformation allowing OTA binding, causing a fluorescence rise due to the increased distance between fluorophore and quencher. We used real-time PCR equipment for capturing the signal. With this assay, under optimized conditions, the entire process can be completed within 1 h. In addition, the proposed system exhibited a good selectivity for OTA against other mycotoxins (ochratoxin B and aflatoxin M1) and limited interference from aflatoxin B1 and patulin. A wide linear detection range (0.2-2000 µM) was achieved, with LOD = 13 nM, r = 0.9952, and R2 = 0.9904. The aptabeacon was also applied to detect OTA in red wine spiked with the same dilution series. A linear correlation with a LOD = 19 nM, r = 0.9843, and R2 = 0.9708 was observed, with recoveries in the range 63%-105%. Intra- and inter-day assays confirmed its reproducibility. The proposed biosensor, although still being finalized, might significantly facilitate the quantitative detection of OTA in...Continue Reading

References

Jul 11, 2001·Analytical Biochemistry·N HamaguchiM Stanton
Oct 25, 2008·Analytical Chemistry·Jorge A Cruz-Aguado, Gregory Penner
Nov 6, 2008·Journal of Agricultural and Food Chemistry·Jorge A Cruz-Aguado, Gregory Penner
Jul 21, 2010·Biosensors & Bioelectronics·Hua KuangShuifang Zhu
Dec 23, 2011·International Journal of Food Microbiology·S M SanzaniC Fanelli
Dec 25, 2012·Biosensors & Bioelectronics·Wei MaChuanlai Xu
Nov 8, 2013·Toxins·Amina RhouatiJean-Louis Marty

❮ Previous
Next ❯

Citations

Jun 18, 2016·Analytical and Bioanalytical Chemistry·Morteza Alinezhad NameghiSeyed Mohammad Taghdisi
Aug 12, 2016·Toxins·Simona Marianna SanzaniCinzia Montemurro
Jul 8, 2016·Toxins·Frantisek MalirJakub Toman
Oct 1, 2020·Analytica Chimica Acta·Elisa SantovitoGiuseppina Avantaggiato

❮ Previous
Next ❯

Methods Mentioned

BETA
biosensors
PCR

Software Mentioned

Graph
iCycler iQTM
Rad Real time Detection System
Bio

Related Concepts

Trending Feeds

COVID-19

Coronaviruses encompass a large family of viruses that cause the common cold as well as more serious diseases, such as the ongoing outbreak of coronavirus disease 2019 (COVID-19; formally known as 2019-nCoV). Coronaviruses can spread from animals to humans; symptoms include fever, cough, shortness of breath, and breathing difficulties; in more severe cases, infection can lead to death. This feed covers recent research on COVID-19.

Blastomycosis

Blastomycosis fungal infections spread through inhaling Blastomyces dermatitidis spores. Discover the latest research on blastomycosis fungal infections here.

Nuclear Pore Complex in ALS/FTD

Alterations in nucleocytoplasmic transport, controlled by the nuclear pore complex, may be involved in the pathomechanism underlying multiple neurodegenerative diseases including Amyotrophic Lateral Sclerosis and Frontotemporal Dementia. Here is the latest research on the nuclear pore complex in ALS and FTD.

Applications of Molecular Barcoding

The concept of molecular barcoding is that each original DNA or RNA molecule is attached to a unique sequence barcode. Sequence reads having different barcodes represent different original molecules, while sequence reads having the same barcode are results of PCR duplication from one original molecule. Discover the latest research on molecular barcoding here.

Chronic Fatigue Syndrome

Chronic fatigue syndrome is a disease characterized by unexplained disabling fatigue; the pathology of which is incompletely understood. Discover the latest research on chronic fatigue syndrome here.

Evolution of Pluripotency

Pluripotency refers to the ability of a cell to develop into three primary germ cell layers of the embryo. This feed focuses on the mechanisms that underlie the evolution of pluripotency. Here is the latest research.

Position Effect Variegation

Position Effect Variagation occurs when a gene is inactivated due to its positioning near heterochromatic regions within a chromosome. Discover the latest research on Position Effect Variagation here.

STING Receptor Agonists

Stimulator of IFN genes (STING) are a group of transmembrane proteins that are involved in the induction of type I interferon that is important in the innate immune response. The stimulation of STING has been an active area of research in the treatment of cancer and infectious diseases. Here is the latest research on STING receptor agonists.

Microbicide

Microbicides are products that can be applied to vaginal or rectal mucosal surfaces with the goal of preventing, or at least significantly reducing, the transmission of sexually transmitted infections. Here is the latest research on microbicides.