Rapid and easy determination of morphine in chafing dish condiments with colloidal gold labeling based lateral flow strips

In order to enhance the flavor of chafing dish and increase the attraction of consumers,the poppy shell is reported to be illegally added to the condiments of chafing dish.In this research,a rapid,simple,and convenient method based on the classic immunochromatographic lateral flow strip(LFS)with gold nanoparticles(GNPs)labeling was developed for easy monitoring of morphine(MOP),an effective component of poppy shell.Under optimized conditions,this developed LFS can well realize the detection of target MOP in the condiments of chafing dish in less than 10 min without any complicated pre-treatments.The limit of detection(LOD)can be achieved as low as 0.1 ppb for standard MOP or the MOP spiked condiments of chafing dish.All these results of the research strongly demonstrate that this established LFS method can be successfully applied in practical rapid and accurate on-site screening of poppy shell in condiments of chafing dish.

A Novel Aptamer Lateral Flow Strip for the Rapid Detection of Gram-positive and Gram-negative Bacteria

Diseases caused by foodborne pathogens are one of the main burdens of public health and seriously hinder global social and economic development.In this research,a novel lateral flow strip was successfully constructed for the simultaneous detection of Gram-positive and Gram-negative bacteria based on the fluidity and color aggregation effect of Au nanoparticles(AuNPs).The lateral flow strip first combined ampicillin(AMP)antigens with AuNPs to form AuNPs-AMP antigens as the first recognition molecule.Then vancomycin specifically recognizing Gram-positive bacteria,and aptamers specifi-cally recognizing Gram-negative bacteria were used as the second recognition molecules.Finally,Gram-positive bacteria and Gram-negative bacteria were detected rapidly using this sandwich mode.The strip can rapidly test the samples within 5 min.Using a lateral flow strip detector,the limit of detection was 4 CFU/mL,and the recovery rates in honey samples were 78.2-88.6%.The lateral flow strip constructed can detect foodborne pathogens quickly,accurately and e fficiently,which is of great significance for food safety detection.

A Sandwich-type Lateral Flow Strip Using a Split, Single Aptamer for Point-of-Care Detection of Cocaine

The sandwich-type lateral fl ow assays relying on dual aptamers with high sensitivity and specifi city have been broadly explored.However,it is unlikely to match a pair of specifi c aptamers that can bind a small molecular target(e.g.,cocaine)simultaneously due to the steric hindrance.In response,we herein introduced the strategy of“one divides into two”into the construction of sandwich-type lateral fl ow strip assay(LFSA).Specifi cally,we split a single cocaine-recognizing aptamer into two segments,either of which was conjugated with gold nanoparticles(AuNPs)or labeled with biotin,serving as signal probe and capture probe,respectively.Upon the presence of the target molecule,a ternary sandwich complex comprised of the two halves of the aptamer and the target formed.Such sandwich-type LFSA exhibited an excellent nonlinear logarithmic response in the range from 10μmol/L to 5 mmol/L with R^(2)=0.9994.The sensitive on-site detection of cocaine in artifi cial biological samples including urine and sweat was achieved within 15 min,with the visual limit of detection as low as 50μmol/L for urine and 200μmol/L for sweat,and the recovery rates of 83.6-107.4%.

Novel ractopamine–protein carrier conjugation and its application to the lateral flow strip test for ractopamine detection in animal feed

In this work,a novel conjugate of ractopamine and bovine serum albumin(RAC–BSA)has been developed via the Mannich reaction,with a mole coupling ratio for RAC–BSA of 9:1.The proposed conjugation method provides a simple and one-step method with the use of fewer reagents compared with other conjugation methods for competitive immunoassays.RAC–BSA conjugation was used to fabricate a competitive lateral flow strip test for RAC detection in animal feed.For sample preparation,RAC was spiked in swine feed purchased from the local markets in Thailand,and methanol and running buffer at a volume ratio of 10:90 was used as extraction buffer.The procedures for sample preparation were completed within 25 min.Under optimal conditions,the limit of detection(LOD),assessed by the naked eye within 5 min,was found to be 1 ng/g.A semi-quantitative analysis was also conducted using a smart phone and computer software,with a linearity of 0.075–0.750 ng/g,calculated LOD of 0.10 ng/g,calculated limit of quantitation of 0.33 ng/g,and good correlation of 0.992.The recoveries were found in the range of 96.4%–103.7%with a relative standard deviation of 2.5%–3.6%for intra-and inter-assays.Comparison of the results obtained by the strip test with those obtained by enzyme-linked immunosorbent assay had a good agreement in terms of accuracy.Furthermore,this strip test exhibited highly specific RAC detection without cross reactivity with related compounds.Therefore,the RAC–BSA conjugation via the Mannich reaction can be accepted as a one-step and easy conjugation method and applied to the competitive lateral flow strip test.

Quantum Dot Nanobeads-Labelled Lateral Flow Immunoassay Strip for Rapid and Sensitive Detection of Salmonella Typhimurium Based on Strand Displacement Loop-Mediated Isothermal Amplification

Rapid,sensitive,point-of-care detection of pathogenic bacteria is important for food safety.In this study,we developed a novel quantum dot nanobeads-labelled lateral flow immunoassay strip(QBs-labelled LFIAS)combined with strand displacement loop-mediated isothermal amplification(SD-LAMP)for quantitative Salmonella Typhimurium(ST)detection.Quantum dot nanobeads(QBs)served as fluorescence reporters,providing good detection efficiency.The customizable strand displacement(SD)probe was used in LAMP to improve the specificity of the method and prevent by-product capture.Detection was based on a sandwich immunoassay.A fluorescence strip reader measured the fluorescence intensity(FI)of the test(T)line and control(C)line.The linear detection range of the strip was 10^(2)–10^(8) colony forming units(CFU)·mL^(-1).The visual limit of detection was 10^(3) CFU·mL^(-1),indicating that the system was ten-fold more sensitive than AuNPs-labelled test strips.ST specificity was analyzed in accordance with agarose gel outputs of polymerase chain reaction(PCR)and SD-LAMP.We detected ST in foods with an acceptable recovery of 85%–110%.The method is rapid,simple,almost equipment-free,and suitable for bacterial detection in foods and for clinical diagnosis.

Filtration assisted pretreatment for rapid enrichment and accurate detection of Salmonella in vegetables

Rapid detection of target foodborne pathogens plays more and more significant roles in food safety,which requires the efficiency,sensitivity,and accuracy.In this research,we proposed a new st rategy of isothermal-molecular-amplification integrated with lateral-flow-strip for rapid detection of Salmonella without traditional enrichment-culture.Th e designed syringe-assisted-filtration can contribute to simultaneous collection and concentration of target bacterium from vegetable samples in just 3 min,resolving the drawbacks of traditional random sampling protocols.After simple and convenient ultrasonication,samples can be directly amplified at 39℃ in 25 min and the amplicons are qualitatively and quantitatively analyzed with the designed lateral-flow-strip in 5 min.Finally,satisfied results have been achieved within 40 min,which greatly improve the efficiency while the accuracy is also guaranteed.Furthermore,all detection steps can be completed under instrument-free conditions.This method will hold great promise for target pathogen detection in the resource-limited district,or for emergency on-site identification.

Dual-readout test strips platform for portable and highly sensitive detection of alkaline phosphatase in human serum samples

In this work,a very simple dual-readout lateral flow test strip(LFTS)platform was developed for sensitive detection of alkaline phosphatase(ALP)based on a portable device.In this assay,quantum dots(QDs)conjugated with bovine serum albumin(QDs-BSA)were chosen as fluorescence signal labels.In the absence of ALP,MnO_(2)nanosheets aggregate on the test line and exhibit an obvious brown color,which can be observed by naked eyes to realize semi-qualitative analysis.Meanwhile,fluorescence intensity of QDs-BSA can also be effectively quenched by MnO_(2)nanosheets due to inner-filter effect.Correspondingly,in the presence of ALP,ALP can catalyze the hydrolysis of ascorbic acid 2-phosphate(AAP)to generate L-ascorbic acid(AA),which can reduce MnO_(2)into Mn^(2+),accompanying with the obvious fluorescence recovery of the QDs.By simply monitoring the change of colorimetric and fluorescent signal on the test line,trace amount of ALP can be quantitatively detected.Under the optimal conditions,measurable evaluation of ALP was reached in a linear range from 1 U/L to 20 U/L with a detection limit of 0.7 U/L based on fluorescence signal.Furthermore,this colorimetric/fluorescent dual-readout assay was successfully applied to monitor ALP in human serum samples,showing its great potential as a point of care biosensor for clinical diagnosis.