An Automated Magnetic Separation Device Coupled with a Fluorescent Biosensor for Detection of Antibiotic Residues

2021 ◽  
Vol 64 (1) ◽  
pp. 23-30
Author(s):  
Aoming Liang ◽  
Yafang Shen ◽  
Yawen He ◽  
Jianping Wang ◽  
Yanbin Li
Keyword(s):  
Magnetic Separation ◽  
Rapid Detection ◽  
Magnetic Beads ◽  
Limit Of Detection ◽  
Sample Pretreatment ◽  
List Type ◽  
Antibiotic Residues ◽  
Separation Device ◽  
Fluorescent Biosensor ◽  
Automated Separation

HighlightsA practical magnetic separation device was designed, fabricated, and evaluated for enrofloxacin detection.Coupled with a fluorescent biosensor, the device could automatically process a sample in 50 min.The device performed incubation and magnetic separation using a pipette method.The device has the advantages of low-cost and feasibility for on-site detection.Abstract. Antibiotic residues have been a continuing concern in food safety, raising a great issue in human health. For rapid detection of antibiotics, an automated device was developed that can capture and separate a target analyte based on immunomagnetic beads. This automated separation device is suitable for separating the magnetic beads in a preprocessing step, with liquid transfer and magnetic enrichment functions. The device was combined with a fluorescent biosensor to simplify the cumbersome pretreatment of enrofloxacin. In our experiments, enrofloxacin in water samples was used as the detection object, and the entire process could be completed in less than 50 min with automated operation. The lower limit of detection reached 54 ng mL-1 (S/N = 3). The fluorescent biosensor has been enhanced with this automated separation device for more sensitive rapid detection of antibiotic residues in the food supply chain and environment. Keywords: Antibiotic detection, Automation, Fluorescent biosensor, Immunomagnetic separation, Sample pretreatment.

scholarly journals Rapid detection of cocaine using aptamer-based biosensor on an evanescent wave fibre platform

2018 ◽  
Vol 5 (10) ◽  
pp. 180821 ◽  
Author(s):  
Yong Qiu ◽  
Yunfei Tang ◽  
Bing Li ◽  
Miao He
Keyword(s):  
Evanescent Wave ◽  
Rapid Detection ◽  
Magnetic Beads ◽  
Feasible Solution ◽  
Limit Of Detection ◽  
Significant Loss ◽  
Saturation Model ◽  
First Order ◽  
First Order Kinetics ◽  
Cocaine Detection

The rapid detection of cocaine has received considerable attention because of the instantaneous and adverse effects of cocaine overdose on human health. Aptamer-based biosensors for cocaine detection have been well established for research and application. However, reducing the analytic duration without deteriorating the sensitivity still remains as a challenge. Here, we proposed an aptamer-based evanescent wave fibre (EWF) biosensor to rapidly detect cocaine in a wide working range. At first, the aptamers were conjugated to complementary DNA with fluorescence tag and such conjugants were then immobilized on magnetic beads. After cocaine was introduced to compete against the aptamer-DNA conjugants, the released DNA in supernatant was detected on the EWF platform. The dynamic curves of EWF signals could be interpreted by the first-order kinetics and saturation model. The semi-log calibration curve covered a working range of 10–5000 µM of cocaine, and the limit of detection was approximately 10.5 µM. The duration of the full procedure was 990 s (16.5 min), and the detection interval was 390 s (6.5 min). The specified detection of cocaine was confirmed from four typical pharmaceutic agents. The analysis was repeated for 50 cycles without significant loss of sensitivity. Therefore, the aptamer-based EWF biosensor is a feasible solution to rapidly detect cocaine.

scholarly journals A novel one-day phage-based test for rapid detection and enumeration of viable Mycobacterium avium subsp. paratuberculosis in cows’ milk

2020 ◽  
Vol 104 (21) ◽  
pp. 9399-9412
Author(s):  
Antonio C. G. Foddai ◽  
Irene R. Grant
Keyword(s):  
Mycobacterium Avium ◽  
Potential Application ◽  
Rapid Detection ◽  
Magnetic Beads ◽  
Limit Of Detection ◽  
Johne's Disease ◽  
Johne’S Disease ◽  
Qpcr Assay ◽  
Specific Test ◽  
Correct Orientation

Abstract Bacteriophage-based methods for the rapid detection of viable Mycobacterium avium subsp. paratuberculosis (MAP) in veterinary specimens are a recent addition to the Johne’s disease diagnostic toolbox. Here, we report the use of D29 mycobacteriophage-coated tosylactivated paramagnetic beads to capture and concentrate MAP cells from samples (termed phagomagnetic separation, PhMS) and then naturally lyse viable MAP cells (from the inside out) to provide DNA for IS900 qPCR purposes. Transmission electron microscopy confirmed that D29 phages had bound to beads in the correct orientation and that the phage-coated beads captured MAP cells from a suspension. During test optimization, conventional IS900 PCR results were used to subjectively assess the effect of different phage:bead coating ratios, differing amounts of coated beads during PhMS, optimal incubation time post-PhMS to obtain maximal MAP DNA, and the potential benefit of a brief heat shock (55 °C/1 min) prior to IS900 TaqMan qPCR. The limit of detection 50% (LOD50%) of the optimised PhMS-qPCR assay was 10.00 MAP cells/50 ml milk (95% CI 1.20–82.83). Finally, in order to demonstrate the new assay’s ability to detect viable MAP in naturally contaminated milk, bulk tank milk samples from 100 dairy farms were tested. Forty-nine (49%) of these tested PhMS-qPCR-positive, with viable MAP numbers detected ranging from 3–126 MAP/50 ml. The novel PhMS-qPCR assay is a sensitive, specific and easy-to-apply phage-based assay for viable MAP, with potential application for milk surveillance or diagnosis of Johne’s disease. Key points • Phage-coated magnetic beads could capture, concentrate and lyse MAP cells from milk. • PhMS-qPCR assay proved to be a rapid, sensitive and specific test for viable MAP. • A potential application of PhMS-qPCR assay for milk surveillance was demonstrated.

High Performance Microfluidic-Based DNA Isolation Chip

2013 ◽  
Author(s):  
Jeff Darabi
Keyword(s):  
Magnetic Separation ◽  
High Performance ◽  
Magnetic Beads ◽  
Biological Entity ◽  
Separation Device ◽  
Separation Systems ◽  
Multiple Sample ◽  
Downstream Analysis ◽  
Biological Entities ◽  
The Magnetic Field

Magnetic separation is one of the effective ways to separate specific biological entities such as DNA/RNA, bacteria, and cells from their native environment for subsequent downstream analysis. The process involves the labeling of the desired biological entity with magnetic beads followed by separating the tagged entities via a magnetic separation device. In conventional tube-based magnetic separation, magnetically labeled biological entities are retained on the inner wall of the tube by applying an external magnet, while the supernatant is decanted off. Removing the tube from the magnetic field enables resuspension of the target entity. Although widely used, there are limitations to the conventional magnetic separation method. For example, there is a significant sample loss due to multiple sample handling, washing, and transfer. In addition, manual magnetic separation systems are labor intensive and their effectiveness is user-dependent.

Highly efficient colorimetric detection of ATP utilizing a split aptamer target binding strategy and superior catalytic activity of graphene oxide–platinum/gold nanoparticles

RSC Advances ◽  
2015 ◽  
Vol 5 (92) ◽  
pp. 75746-75752 ◽  
Author(s):  
Siqi Zhang ◽  
Kun Wang ◽  
Jiali Li ◽  
Zhenyu Li ◽  
Ting Sun
Keyword(s):  
Gold Nanoparticles ◽  
Graphene Oxide ◽  
Catalytic Activity ◽  
Magnetic Separation ◽  
Rapid Detection ◽  
Magnetic Beads ◽  
Specific Binding ◽  
Colorimetric Detection ◽  
Colored Product ◽  
Target Binding

The specific binding of ATP and its aptamer linked the split aptamer-modified GO/PDDA/PtAuNPs and magnetic beads together. Using magnetic separation, TMB was catalyzed into a colored product by nanocomposites, which enabled rapid detection of ATP.

scholarly journals Urine lipoarabinomannan in HIV uninfected, smear negative, symptomatic TB patients: effective sample pretreatment for a sensitive immunoassay and mass spectrometry

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Anita G. Amin ◽  
Prithwiraj De ◽  
Barbara Graham ◽  
Roger I. Calderon ◽  
Molly F. Franke ◽  
...  
Keyword(s):  
Limit Of Detection ◽  
Sample Pretreatment ◽  
Positive Association ◽  
Proteinase K ◽  
Clinical Samples ◽  
Sputum Smear ◽  
Smear Negative ◽  
Tuberculostearic Acid ◽  
Negative Sputum ◽  
Adult Culture

AbstractOur study sought to determine whether urine lipoarabinomannan (LAM) could be validated in a sample cohort that consisted mainly of HIV uninfected individuals that presented with tuberculosis symptoms. We evaluated two tests developed in our laboratory, and used them on clinical samples from Lima, Peru where incidence of HIV is low. ELISA analysis was performed on 160 samples (from 140 adult culture-confirmed TB cases and 20 symptomatic TB-negative child controls) using 100 μL of urine after pretreatment with Proteinase K. Two different mouse monoclonal antibodies-CS35 and CHCS9-08 were used individually for capture of urine LAM. Among cases, optical density (OD450) values had a positive association with higher bacillary loads. The 20 controls had negative values (below the limit of detection). The assay correctly identified all samples (97–100% accuracy confidence interval). For an alternate validation of the ELISA results, we analyzed all 160 urine samples using an antibody independent chemoanalytical approach. Samples were called positive only when LAM surrogates—tuberculostearic acid (TBSA) and d-arabinose (d-ara)—were found to be present in similar amounts. All TB cases, including the 40 with a negative sputum smear had LAM in detectable quantities in urine. None of the controls had detectable amounts of LAM. Our study shows that urinary LAM detection is feasible in HIV uninfected, smear negative TB patients.

scholarly journals Identification and dietary exposure assessment of tetracycline and penicillin residues in fluid milk, yogurt, and labneh: A cross-sectional study in Lebanon

2019 ◽  
Vol 12 (4) ◽  
pp. 527-534 ◽  
Author(s):  
Suzanne Kabrite ◽  
Christelle Bou-Mitri ◽  
Jessy El Hayek Fares ◽  
Hussein F. Hassan ◽  
Jocelyne Matar Boumosleh
Keyword(s):  
Human Health ◽  
Dairy Products ◽  
Detection Rate ◽  
Limit Of Detection ◽  
Human Health Risk ◽  
Daily Intake ◽  
Mean Value ◽  
Enzyme Linked Immunosorbent Assay ◽  
Antibiotic Residues ◽  
Mean Values

Background and Aim: The safety and quality of dairy products are considered to be of significant importance to human health. Although antimicrobial drugs are essential for disease treatment in modern medicine, the use of these drugs can have undesired consequences for human and animal health. This study aimed to investigate the presence of tetracycline and penicillin residues in raw, pasteurized, and UHT cow's milk of different fat contents, as well as in the dairy products yogurt and labneh, a traditional Lebanese product. Materials and Methods: A total of 44 samples, 4 raw, 9 UHT, 9 pasteurized milk, 10 yogurt, and 12 labneh samples from common local brands available in the Lebanese market were collected from Keserwan regions in May 2016. Tetracycline and penicillin residues were determined using a competitive enzyme-linked immunosorbent assay (ELISA) technique. Results: The mean values for tetracycline and penicillin were all below the limit of detection (LOD) of the ELISA kit of a maximum standard concentration of 1.80 μg/kg and 4.00 μg/kg, respectively. All samples tested positive for antibiotic residues. The detection rate for tetracycline in milk (n=22) samples was 86.4% with a mean residues value of 1.16±0.70 μg/kg. The detection rate of tetracycline in labneh (n=12) and yogurt (n=10) samples was 50% for each with a mean value of 1.76±0.40 μg/kg and 0.63±0.12 μg/kg, respectively. As for penicillin residues, 90.9% of the milk (n=22) samples tested positive with a mean value of 0.52±0.25 μg/kg. The detection rate in labneh (n=12) and yogurt (n=10) samples was 0% for penicillin residues, where mean values were all below the LOD (<1.25 μg/kg) for these dairy products. None of the samples exceeded the maximum residue levels. The estimated dietary intake (EDI) for tetracycline and penicillin residues for all dairy products is 2.09 ng/kg body weight (BW)/day resulting in 0.007% of the acceptable daily intake (ADI) and 1.83 ng/kg BW/day resulting in 0.006% of the ADI, respectively. Conclusion: All EDI values were below the ADI set for each antibiotic residue and do not exceed relevant toxicological reference values. However, concerns might still be present from consumption of other animal food products containing residues. Moreover, the long-term exposure to such residues is still unknown as a result of bioaccumulation; it is a challenging process to determine the actual dietary consumption of foods containing antibiotic residues; hence, the human health risk cannot be easily predicted.

scholarly journals A novel fluorescent biosensor based on dendritic DNA nanostructure in combination with ligase reaction for ultrasensitive detection of DNA methylation

2019 ◽  
Vol 17 (1) ◽  
Author(s):  
Shu Zhang ◽  
Jian Huang ◽  
Jingrun Lu ◽  
Min Liu ◽  
Yan Li ◽  
...  
Keyword(s):  
Dna Methylation ◽  
Limit Of Detection ◽  
Fluorescent Dye ◽  
Fluorescence Signal ◽  
Target Sequence ◽  
Capture Probe ◽  
Detection Range ◽  
Fluorescent Biosensor ◽  
Dna Nanostructure ◽  
Reporter Probe

Background DNA methylation detection is indispensable for the diagnosis and prognosis of various diseases including malignancies. Hence, it is crucial to develop a simple, sensitive, and specific detection strategy. Methods A novel fluorescent biosensor was developed based on a simple dual signal amplification strategy using functional dendritic DNA nanostructure and signal-enriching polystyrene microbeads in combination with ligase detection reaction (LDR). Dendritic DNA self-assembled from Y-DNA and X-DNA through enzyme-free DNA catalysis of a hairpin structure, which was prevented from unwinding at high temperature by adding psoralen. Then dendritic DNA polymer labeled with fluorescent dye Cy5 was ligated with reporter probe into a conjugate. Avidin-labeled polystyrene microbeads were specifically bound to biotin-labeled capture probe, and hybridized with target sequence and dendritic DNA. LDR was triggered by adding Taq ligase. When methylated cytosine existed, the capture probe and reporter probe labeled with fluorescent dye perfectly matched the target sequence, forming a stable duplex to generate a fluorescence signal. However, after bisulfite treatment, unmethylated cytosine was converted into uracil, resulting in a single base mismatch. No fluorescence signal was detected due to the absence of duplex. Results The obtained dendritic DNA polymer had a large volume. This method was time-saving and low-cost. Under the optimal experimental conditions using avidin-labeled polystyrene microbeads, the fluorescence signal was amplified more obviously, and DNA methylation was quantified ultrasensitively and selectively. The detection range of this sensor was 10−15 to 10−7 M, and the limit of detection reached as low as 0.4 fM. The constructed biosensor was also successfully used to analyze actual samples. Conclusion This strategy has ultrasensitivity and high specificity for DNA methylation quantification, without requiring complex processes such as PCR and enzymatic digestion, which is thus of great value in tumor diagnosis and biomedical research.

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