scholarly journals Multiplex Lateral Flow Immunoassay for the Detection of Expanded-Spectrum Hydrolysis and CTX-M Enzymes

Diagnostics ◽  
2022 ◽  
Vol 12 (1) ◽  
pp. 190
Author(s):  
Christian Moguet ◽  
Camille Gonzalez ◽  
Thierry Naas ◽  
Stéphanie Simon ◽  
Hervé Volland
Keyword(s):  
Room Temperature ◽  
Low Cost ◽  
Lateral Flow ◽  
Lateral Flow Immunoassay ◽  
Bacterial Isolates ◽  
Negative Results ◽  
Single Colony ◽  
Extraction Buffer ◽  
Acinetobacter Spp ◽  
Test Result

Background: Early detection of expanded-spectrum cephalosporinase (ESC) hydrolyzing ß-lactamases is essential for antibiotic stewardship. Here we have developed a multiplex lateral flow immunoassay (LFIA) that detects cefotaxime-hydrolyzing activity as well as the most prevalent ESC-hydrolyzing ß-lactamases: the CTX-M-like. Methods: The Rapid LFIA ESC test was evaluated retrospectively on 188 (139 Enterobacterales, 30 Pseudomonas spp. and 14 Acinetobacter spp.) agar-grown bacterial isolates with well-characterized ß-lactamase content. One single colony was resuspended in 150 µL extraction buffer containing cefotaxime, incubated at room temperature for 30 min prior to loading on the LFIA for reading within 10 min. Results: Out of the 188 isolates, all 17 that did not express a β-lactamase hydrolyzing cefotaxime gave negative results, and all 171 isolates expressing a β-lactamase known to hydrolyze cefotaxime, gave a positive test result. In addition, all 86 isolates expressing a CTX-M-variant belonging to one of the five CTX-M-subgroups were correctly identified. The sensitivity and specificity was 100% for both tests. Conclusions: The results showed that the multiplex LFIA was efficient, fast, low cost and easy to implement in routine laboratory work for the confirmation of ESC hydrolyzing activity and the presence of CTX-M enzymes.

scholarly journals Identification of donkey meat in foods using species-specific PCR combined with lateral flow immunoassay

RSC Advances ◽  
2019 ◽  
Vol 9 (46) ◽  
pp. 26552-26558 ◽  
Author(s):  
Liangjuan Zhao ◽  
Marti Z. Hua ◽  
Shenmiao Li ◽  
Jinyu Liu ◽  
Wenjie Zheng ◽  
...  
Keyword(s):  
Low Cost ◽  
Lateral Flow ◽  
Lateral Flow Immunoassay ◽  
Specific Detection ◽  
Chain Reaction ◽  
Specific Pcr ◽  
Highly Sensitive ◽  
Polymerase Chain ◽  
Species Specific ◽  
Government Laboratories

The developed species-specific polymerase chain reaction-lateral flow immunoassay (PCR-LFI) method allows the rapid, low-cost, highly sensitive and specific detection of donkey DNA for meat authentication, adopted by government laboratories.

scholarly journals COVALENT CONJUGATION OF ANTIBODY AND GOLD NANOPARTICLE FOR DEVELOPMENT OF LATERAL FLOW IMMUNOASSAY TEST STRIP

2018 ◽  
Vol 54 (4A) ◽  
pp. 323
Author(s):  
Truong Quoc Phong
Keyword(s):  
Gold Nanoparticle ◽  
Room Temperature ◽  
Limit Of Detection ◽  
Coupling Reaction ◽  
Test Strip ◽  
Lateral Flow ◽  
Lateral Flow Immunoassay ◽  
Research Fields ◽  
Antibody Conjugates ◽  
Antigen Antibody

Nanotechnology is one of the fastest growing technologies in this era. Gold nanoparticle (AuNP) based immunoassays have been performed on the basis of antigen-antibody interaction using AuNP antibody conjugates. Lateral flow immunoassays(LFA) which are also based on AuNP antibody conjugates are useful innovation in nanotechnology and widely applied in medicine and research fields. However, there are some limitations of the present LFA kits such as sensitivity and stability. In the study, we showed the result of covalent conjugation of anti-rotavirus antibody and AuNP for generating a lateral flow immunoassay strip to detect rotavirus in fecal samples. The suitable conditions for coating polyethylene glycol (PEG) on the surface of AuNP were 10.0 mM PEG for 3 hours at room temperature (25 oC). Optimized conditions for covalent conjugation of antibody and AuNP were pH 4.0, 0.1 mg antibody/conjugate, 0.01 mM reactant EDC/NHS [1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride/(N-hydroxy sulfosuccinimide]. The coupling reaction was carried out at room temperature for 90 min. The conjugate pad, antibody immobilized nitrocellulose membrane strip were created with investigated conditions for generating an LFA test strip. The limit of detection of LFA test strip was determined by 1.6 × 105 virus particles/ml, three times lower than that of Rotaclone kit (UK). The generated strip could be used to detect rotavirus in fecal sample of patient. 

scholarly journals A Lateral Flow Immunoassay for the Rapid Identification of CTX-M-Producing Enterobacterales from Culture Plates and Positive Blood Cultures

Diagnostics ◽  
2020 ◽  
Vol 10 (10) ◽  
pp. 764 ◽  
Author(s):  
Sandrine Bernabeu ◽  
Kayaththiry Caroline Ratnam ◽  
Hervé Boutal ◽  
Camille Gonzalez ◽  
Anaïs Vogel ◽  
...  
Keyword(s):  
Clinical Setting ◽  
Clinical Microbiology ◽  
Rapid Identification ◽  
Blood Cultures ◽  
Lateral Flow ◽  
Lateral Flow Immunoassay ◽  
Gram Negative ◽  
Negative Results ◽  
Esbl Producers ◽  
Group 1

We have developed a lateral flow immunoassay (LFIA), named NG-Test CTX-M MULTI (NG-Test), to detect group 1, 2, 8, 9, 25 CTX-M producers from agar plates and from positive blood cultures in less than 15 min. The NG-Test was validated retrospectively on 113 well-characterized enterobacterial isolates, prospectively on 102 consecutively isolated ESBL-producers from the Bicêtre hospital and on 100 consecutive blood cultures positive with a gram-negative bacilli (GNB). The NG-Test was able to detect all CTX-M producers grown on the different agar plates used in clinical microbiology laboratories. No false positive nor negative results were observed. Among the 102 consecutive ESBL isolates, three hyper mucous isolates showed an incorrect migration leading to invalid results (no control band). Using an adapted protocol, the results could be validated. The NG-Test detected 99/102 ESBLs as being CTX-Ms. Three SHV producers were not detected. Among the 100 positive blood cultures with GNB tested 10/11 ESBL-producers were detected (8 CTX-M-15, 2 CTX-M-27). One SHV-2-producing-E. cloacae was missed. The NG-Test CTX-M MULTI showed 100% sensitivity and specificity with isolates cultured on agar plates and was able to detect 98% of the ESBL-producers identified in our clinical setting either from colonies or from positive blood cultures.

A paper microfluidics-based fluorescent lateral flow immunoassay for point-of-care diagnostics of non-communicable diseases

The Analyst ◽  
2019 ◽  
Vol 144 (21) ◽  
pp. 6291-6303 ◽  
Author(s):  
Satheesh Natarajan ◽  
Fengmei Su ◽  
Joseph Jayaraj ◽  
Malay I. Iesh Shah ◽  
Youju Huang
Keyword(s):  
Point Of Care ◽  
Low Cost ◽  
Communicable Diseases ◽  
Lateral Flow ◽  
Lateral Flow Immunoassay ◽  
Paper Microfluidics ◽  
Non Communicable Diseases ◽  
Point Of Care Diagnostics

An easy-to-use, low-cost and portable fluorescent lateral flow immunoassay based on paper microfluidics was developed for the point-of-care diagnosis of non-communicable diseases.

Green Synthesis of 10 nm Gold Nanoparticles via Seeded-Growth Method and its Conjugation Properties on Lateral Flow Immunoassay

2013 ◽  
Vol 686 ◽  
pp. 8-12 ◽  
Author(s):  
Rabizah Makhsin Siti ◽  
Abdul Razak Khairunisak ◽  
Abdul Aziz Azlan ◽  
Rahmah Noordin
Keyword(s):  
Gold Nanoparticles ◽  
Low Cost ◽  
Hydroxylamine Hydrochloride ◽  
Trisodium Citrate ◽  
Reducing Agent ◽  
Lateral Flow ◽  
Lateral Flow Immunoassay ◽  
Seeded Growth ◽  
Vis Spectroscopy ◽  
Growth Method

In this work, 10 nm gold nanoparticles (AuNPs) was successfully synthesized via seeded-growth method. The green chemically synthesis of this AuNPs becomes attractive because the growth process does not involve heat. Moreover, this technique has advantages of quick, simple, and low cost process. Sodium borohydrate (NaBH4) was used as a reducing agent while trisodium citrate was used as a source of OH- ions in the seed stage. Hydroxylamine hydrochloride (NH4.3H2O) was used as a slow reducing agent to enlarge 4 nm seeds to 10 nm AuNPs. A 4 ml AuNPs seed was the optimized volume to produce 10 nm AuNPs with great homogeneity and dispersity. A sharp peak of surface plasmon resonance (SPR) measurement at 517 nm proved that 10 nm AuNPs was successfully synthesized via this method. Optical properties of the seeds and grown AuNPs were analyzed using UV-Vis spectroscopy while size and surface morphology were observed using a transmission electron microscopy (TEM). Particle size distribution was measured using Zetasizer. 10 nm AuNPs was then conjugated with streptavidin and goat anti-human IgA. Depending on type of protein, 10 µg/ml of streptavidin and 11.2 µg/ml of goat anti-human IgA were required to conjugate with 10 nm AuNPs. The produced products had binding capability on lateral flow immunoassay (LFI). A few nanometer red-shifted absorption spectrum of 10 nm AuNPs conjugated protein revealed successful conjugation instead of agglomeration. 1% BSA was determined as the optimum concentration to stabilize 10 nm AuNPs conjugated biomolecules.

scholarly journals Evaluation of rapid, cassette immunochromatographic tests in the serological diagnosis of COVID-19

2021 ◽  
pp. 25-37
Author(s):  
Waldemar Rastawicki ◽  
Klaudia Płaza ◽  
Adam Pietrusiński
Keyword(s):  
Low Cost ◽  
Lateral Flow ◽  
Serological Diagnosis ◽  
Igg Antibodies ◽  
Serum Samples ◽  
Serological Tests ◽  
Negative Results ◽  
Lateral Flow Assays ◽  
Low Sensitivity ◽  
Positive Results

Introduction: Lateral flow assays (LFIA) are the technology behind low-cost, simple, rapid and portable detection devices popular in biomedicine. Lately, they are very common used in serodiagnosis of SARS-CoV-2 infections. The aim of the presented study was to assess the usefulness of selected LFIA in serological diagnosis of COVID-19. Methods: The usefulness of seven lateral flow assays in the serodiagnosis of COVID-19 was evaluated (VAZYME, DIAGNOSIS, PCL, INGEZIM, BIOSENSOR, ACCU-TELL, NOVAtest). The study used 107 serum samples obtained from 74 individuals with current SARS-CoV-2 infection confirmed by RT-PCR. The ELISA-IgG (Euroimmun) was used as the reference assay for sensitivity and specificity testing. Results: The highest percentage of positive results was obtained when searching for IgG antibodies with the NOVAtest (40.6%) and DIAGNOSIS (39.2%) sets and the lowest detection for the PCL set - 25.5%. In the case of searching for IgM antibodies in all sets, significantly lower percentages of positive results compared to the IgG class were recorded. In general, all lateral flow assays showed low sensitivity in relation to the Euroimmun ELISA-IgG. The DIAGNOSIS kit (64.5%) was characterized by the highest sensitivity, and the PCL kit was the lowest (38.7%). On the other hand, the specificity of all kits was very high, almost 100% in almost all cases. Conclusions: Lateral flow assays due to their low sensitivity are not suitable for quick diagnosis of the current SARS-CoV-2 infections and cannot be an alternative to genetic or even antigen tests. They may be used only to retrospectively test the presence of IgG antibodies. However, a negative results of LFIA in suspected disease or after vaccination should be confirmed by more sensitive serological tests.

Development of a lateral flow immunoassay strip for rapid detection of IgG antibody against SARS-CoV-2 virus

The Analyst ◽  
2020 ◽  
Vol 145 (15) ◽  
pp. 5345-5352 ◽  
Author(s):  
Tian Wen ◽  
Chao Huang ◽  
Feng-Juan Shi ◽  
Xiao-Yan Zeng ◽  
Tian Lu ◽  
...  
Keyword(s):  
Rapid Detection ◽  
Correct Diagnosis ◽  
Preliminary Test ◽  
Lateral Flow ◽  
Lateral Flow Immunoassay ◽  
Testing Methods ◽  
Igg Antibody ◽  
Au Nps ◽  
Alternative Testing ◽  
Test Result

Rapid and simple LFIA strips based on Au NPs provide a preliminary test result for physicians to make the correct diagnosis of SARS-CoV-2 infections along with alternative testing methods.

The Characterization of Antibiotic Resistance of Bacterial Isolates from Intensive Care Unit Patient Samples in a University Affiliated Hospital in Romania

2019 ◽  
Vol 70 (5) ◽  
pp. 1778-1783
Author(s):  
Andreea-Loredana Golli ◽  
Floarea Mimi Nitu ◽  
Maria Balasoiu ◽  
Marina Alina Lungu ◽  
Cristiana Cerasella Dragomirescu ◽  
...  
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Antibiotic Resistance ◽  
Bacterial Pathogens ◽  
Resistance Rate ◽  
Resistance Pattern ◽  
Bacterial Isolates ◽  
E Coli ◽  
Acinetobacter Spp ◽  
Almost All ◽  
Klebsiella Spp

To determine the resistance pattern of bacterial pathogens involved in infections of the patients aged between 18-64 years, admitted in a ICU from a 1518-bed university-affiliated hospital. A retrospective study of bacterial pathogens was carried out on 351 patients aged between 18-64 years admitted to the ICU, from January to December 2017. In this study there were analysed 469 samples from 351 patients (18-64 years). A total of 566 bacterial isolates were obtained, of which 120 strains of Klebsiella spp. (35.39%%), followed by Nonfermenting Gram negative bacilli, other than Pseudomonas and Acinetobacter (NFB) (75- 22.12%), Acinetobacter spp. (53 - 15.63%), Pseudomonas aeruginosa and Proteus (51 - 15.04%), and Escherichia coli (49 - 14.45%). The most common isolates were from respiratory tract (394 isolates � 69.61%). High rates of MDR were found for Pseudomonas aeruginosa (64.70%), MRSA (62.65%) and Klebsiella spp. (53.33%), while almost all of the isolated NFB strains were MDR (97.33%). There was statistic difference between the drug resistance rate of Klebsiella and E. coli strains to ceftazidime and ceftriaxone (p[0.001), cefuroxime (p[0.01) and to cefepime (p[0.01). The study revealed an alarming pattern of antibiotic resistance in the majority of ICU isolates.

Graphene Modified Molecular Imprinting Electrochemical Sensor for Determining the Content of Dopamine

2019 ◽  
Vol 15 (6) ◽  
pp. 628-634
Author(s):  
Rong Liu ◽  
Jie Li ◽  
Tongsheng Zhong ◽  
Liping Long
Keyword(s):  
Electrochemical Sensor ◽  
Molecular Imprinting ◽  
Neurological Disorders ◽  
Room Temperature ◽  
Low Cost ◽  
Differential Pulse ◽  
Current Response ◽  
Specific Selectivity ◽  
Pulse Voltammetry

Background: The unnatural levels of dopamine (DA) result in serious neurological disorders such as Parkinson’s disease. Electrochemical methods which have the obvious advantages of simple operation and low-cost instrumentation were widely used for determination of DA. In order to improve the measurement performance of the electrochemical sensor, molecular imprinting technique and graphene have always been employed to increase the selectivity and sensitivity. Methods: An electrochemical sensor which has specific selectivity to (DA) was proposed based on the combination of a molecular imprinting polymer (MIP) with a graphene (GR) modified gold electrode. The performance and effect of MIP film were investigated by differential pulse voltammetry (DPV) and cyclic voltammetry (CV) in the solution of 5.0 ×10-3 mol/L K3[Fe(CN)6] and K4[Fe(CN)6] with 0.2 mol/L KCl at room temperature. Results: This fabricated sensor has well repeatability and stability, and was used to determine the dopamine of urine. Under the optimized experiment conditions, the current response of the imprinted sensor was linear to the concentration of dopamine in the range of 1.0×10-7 ~ 1.0×10-5 mol/L, the linear equation was I (µA) = 7.9824+2.7210lgc (mol/L) with the detection limit of 3.3×10-8 mol/L. Conclusion: In this work, a highly efficient sensor for determination of DA was prepared with good sensitivity by GR and great selectivity of high special recognization ability by molecular imprinting membrane. This proposed sensor was used to determine the dopamine in human urine successfully.

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