Quantitative, False Positive, and False Negative Issues for Lateral Flow Immunoassays as Exemplified by Onsite Drug Screens

Wong, Raphael C.; Tse, Harley Y.

doi:10.1007/978-1-59745-240-3_10

Cited by 79 publications

(108 citation statements)

References 23 publications

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“…For example, lateral-flow immunoassays, i.e., immunochromatographies, are already available for on-site testing of illicit drugs, including THC and its metabolites. 7) Genetically engineering of antibodies (antibody engineering) has also generated artificial antibodies with valuable functions. 8,9) For instance, native antibodies (immunoglobulins) can be reduced into much smaller single-chain Fv fragments (scFvs), ca.…”

Section: 6)mentioning

confidence: 99%

Antibody Fragments for On-Site Testing of Cannabinoids Generated <i>via in Vitro</i> Affinity Maturation

Morita

Oyama

Yasuo

et al. 2017

Biological & Pharmaceutical Bulletin

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Law enforcement against illicit use of cannabis and related substances requires rapid, feasible, and reliable tools for on-site testing of cannabinoids. Notably, methods based on cannabinoid-specific antibodies enable efficient screening of multiple specimens. Antibody engineering may accelerate development of modern and robust testing systems. Here, we used in vitro affinity maturation to generate a single-chain Fv fragment (scFv) that recognizes with high affinity the psychoactive cannabinoid, Δ 9 -tetrahydrocannabinol (THC). A mouse monoclonal antibody against THC, Ab-THC#33, with K a 6.2 10 7 M 1 (as Fab fragment) was established by the hybridoma technique. Then, a "wild-type" scFv (wt-scFv) with K a , 1.1 10 7 M 1 was prepared by bacterial expression of a fusion gene combining the V H and V L genes for Ab-THC#33. Subsequently, random point mutations in V H and V L were generated separately, and the resulting products were assembled into mutant scFv genes, which were then phage-displayed. Repeated panning identified a mutant scFv (scFv#m1-36) with 10-fold enhanced affinity (K a 1.1 10 8 M 1 ) for THC, in which only a single conservative substitution (Ser50Thr) was present at the N-terminus of the V H -complementarity-determining region 2 (CDR2) sequence. In competitive enzyme-linked immunosorbent assay (ELISA), the mutant scFv generated dose-response curves with midpoint 0.27 ng/assay THC, which was 3-fold lower than that of wt-scFv. Even higher reactivity with a major THC metabolite, 11-nor-9-carboxy-Δ 9 -tetrahydrocannabinol, indicated that the mutant scFv will be useful for testing not only THC in confiscated materials, but also the metabolite in urine. Indeed, the antibody fragment is potentially suitable for use in advanced on-site testing platforms for cannabinoids.

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Section: 6)mentioning

confidence: 99%

Antibody Fragments for On-Site Testing of Cannabinoids Generated <i>via in Vitro</i> Affinity Maturation

Morita

Oyama

Yasuo

et al. 2017

Biological & Pharmaceutical Bulletin

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“…Therefore, a large number of data points are captured by the CCD camera, which requires a highly efficient computer to accomplish the appropriate image processing. Also, the performance of the image processing algorithm has a great impact on the detection accuracy and CCDs are comparatively expensive, which are not suitable for use in a low budget LFIA test reader system [9].…”

Section: Introductionmentioning

confidence: 99%

“…Therefore, signal processing algorithm for the scanning based LFIA reader system is simpler than image processing. Furthermore, it markedly reduces the computational burden and the overall system cost [9]. Gu et al developed a portable fluorescence reader with an S3C2440AL40 (Samsung, Korea) as a main controller and an AT89C51E2 (Atmel, USA) as an auxiliary controller [7].…”

Section: Introductionmentioning

confidence: 99%

A Lateral Flow Immunochromatographic Assay (LFIA) Strip Reader Based on Scheduler and 8051 IP Core

Wei

Gao

Li³

et al. 2016

Automatika

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Original scientific paperLateral flow immunochromatographic assay (LFIA) testing is essential for accurate detection and diagnoses of diseases and physical conditions. However, the existing LFIA strip reader equipped with high cost of hardware confines its simplicity and portability. Therefore, this study develops a simple, low cost LFIA strip reader comprising 4 major modules -mechanical, optical, processing and control modules. The mechanical module pulls in and out the test strip automatically to be read by the optical module and the data processing module provides the test results by analyzing the data sent by the optical module. All the individual modules are controlled by a control module. To reduce the hardware budget and control complexity, a time-triggered cooperative (TTC) scheduler implemented on an 8051 IP core was chosen as control system. In addition, special, high sensitivity C-reactive protein (CRP) strips with 10 different concentrations were tested to evaluate the performance of the system. Further, a commercial ESEQuant lateral flow reader (QIAGEN, Germany) was tested as a comparative study. The test results show that the proposed reader was stable with a coefficient of variation (CV) factor within 3%. To test the qualitative performance of the system, each of the CRP concentration were examined for 10 times, which indicates that the system has a large dynamic detection range and good detection linearity (R 2 = 0.998). In short, the proposed LFIA strip reader has high potential relative to existing readers, especially in simplicity and cost. Imunokromatografsko testiranje lateralnog toka (LFIA)čitača trake temeljeno na rasporedu i 8051 IP jezgri. Imunokromatografsko testiranje lateralnog toka (LFIA) nužno je za preciznu detekciju i dijagnozu oboljenja te psihičkih stanja. Meutim, postojeći LFIA opremljeni s hardverom visoke cijene limitiraju jednostavnost i prenosivost. Ovo istraživanje razvija jednostavni, niske cijene,čitač traka koji se sastoji od 4 glavna dijelamehanički, optički, procesni i upravljački. Mehanički modul povlači testnu traku automatski kako bi optički modul mogaočitati. Procesni modul analizira podatke dobivene s optičkogčitača. Svaki modula upravlja se upravljačkim modulom. Vremensko ovisno kooperativni raspored implementiran je na 8051 IP jezgri kako bi se smanjili računski zahtjevi. Dodatno, visoko osjetljiva CRP traka s deset različitih koncentracija korištena je u svrhe evaluacije sustava. Rezultati su usporeeni s komercijalnimčitačem lateralnog toka ESEQuant (QIAGEN, Njemačka). Rezultati pokazuju da je predložena metoda stabilna s koeficijentom varijacije unutar 3%. Kako bi se kvalitativno testirao sustav, svaka od CRP koncentracija testirana je deset puta, što ukazuje da sustav veliki dinamički raspon detekcije te dobru linearnost detekcije (R 2 = 0.998). Predloženi LFIAčitač traka ima dosta potencijala u usporedbi s postoječimčitačima, posebno u smislu jednostavnosti i cijene.

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“…Compared with conventional liquid handling systems, such miniaturized technologies have enabled the manipulation of small volume liquids with unprecedented resolution (picoliter or less) 8 , automation 9 and parallelism 10 . However, due to various fabrication, integration and reliability challenges, few handheld self-contained microfluidic systems capable of complex liquid handling exist in the market today except for capillary-driven microfluidics 11 . Most labon-a-chip systems still rely on bulky off-chip infrastructures such as compressed pressure sources, syringe pumps and computers to achieve their liquid manipulation functions.…”

Section: Introductionmentioning

confidence: 99%

A smartphone controlled handheld microfluidic liquid handling system

Guan

et al. 2014

Lab Chip

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Microfluidics and lab-on-a-chip technologies have made it possible to manipulate small volume liquids with unprecedented resolution, automation and integration. However, most current microfluidic systems still rely on bulky off-chip infrastructures such as compressed pressure sources, syringe pumps and computers to achieve complex liquid manipulation functions. Here, we present a handheld automated microfluidic liquid handling system controlled by a smartphone, which is enabled by combining elastomeric on-chip valves and a compact pneumatic system. As a demonstration, we show that the system can automatically perform all the liquid handling steps of a bead-based sandwich immunoassay on a multi-layer PDMS chip without any human intervention. The footprint of the system is 6 × 10.5 × 16.5cm, and the total weight is 829g including battery. Powered by a 12.8V 1500mAh Li battery, the system consumed 2.2W on average during the immunoassay and lasted for 8.7 hrs. This handheld microfluidic liquid handling platform is generally applicable to many biochemical and cell-based assays requiring complex liquid manipulation and sample preparation steps such as FISH, PCR, flow cytometry and nucleic acid sequencing. In particular, the integration of this technology with read-out biosensors may help enable the realization of the long-sought Tricorder-like handheld in-vitro diagnostic (IVD) systems.

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Quantitative, False Positive, and False Negative Issues for Lateral Flow Immunoassays as Exemplified by Onsite Drug Screens

Cited by 79 publications

References 23 publications

Antibody Fragments for On-Site Testing of Cannabinoids Generated <i>via in Vitro</i> Affinity Maturation

Antibody Fragments for On-Site Testing of Cannabinoids Generated <i>via in Vitro</i> Affinity Maturation

A Lateral Flow Immunochromatographic Assay (LFIA) Strip Reader Based on Scheduler and 8051 IP Core

A smartphone controlled handheld microfluidic liquid handling system

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