scholarly journals Lateral Flow Immunoassay of SARS-CoV-2 Antigen with SERS-Based Registration: Development and Comparison with Traditional Immunoassays

Biosensors ◽  
2021 ◽  
Vol 11 (12) ◽  
pp. 510
Author(s):  
Kseniya V. Serebrennikova ◽  
Nadezhda A. Byzova ◽  
Anatoly V. Zherdev ◽  
Nikolai G. Khlebtsov ◽  
Boris N. Khlebtsov ◽  
...  
Keyword(s):  
Pathogen Detection ◽  
Specific Antigen ◽  
Lateral Flow ◽  
Nucleic Acid Amplification ◽  
Detection Methods ◽  
Lateral Flow Immunoassay ◽  
Enzyme Linked Immunosorbent Assay ◽  
Highly Qualified ◽  
Photometric Detection ◽  
Low Sensitivity

The current COVID-19 pandemic has increased the demand for pathogen detection methods that combine low detection limits with rapid results. Despite the significant progress in methods and devices for nucleic acid amplification, immunochemical methods are still preferred for mass testing without specialized laboratories and highly qualified personnel. The most widely used immunoassays are microplate enzyme-linked immunosorbent assay (ELISA) with photometric detection and lateral flow immunoassay (LFIA) with visual results assessment. However, the disadvantage of ELISA is its considerable duration, and that of LFIA is its low sensitivity. In this study, the modified LFIA of a specific antigen of the causative agent of COVID-19, spike receptor-binding domain, was developed and characterized. This modified LFIA includes the use of gold nanoparticles with immobilized antibodies and 4-mercaptobenzoic acid as surface-enhanced Raman scattering (SERS) nanotag and registration of the nanotag binding by SERS spectrometry. To enhance the sensitivity of LFIA-SERS analysis, we determined the optimal compositions of SERS nanotags and membranes used in LFIA. For benchmark comparison, ELISA and conventional colorimetric LFIA were used with the same immune reagents. The proposed method combines a low detection limit of 0.1 ng/mL (at 0.4 ng/mL for ELISA and 1 ng/mL for qualitative LFIA) with a short assay time equal to 20 min (at 3.5 h for ELISA and 15 min for LFIA). The results obtained demonstrate the promise of using the SERS effects in membrane immuno-analytical systems.

scholarly journals Comparative Analysis of the Euroimmun CXCL13 Enzyme-Linked Immunosorbent Assay and the ReaScan Lateral Flow Immunoassay for Diagnosis of Lyme Neuroborreliosis

2020 ◽  
Vol 58 (9) ◽  
Author(s):  
Katharina Ziegler ◽  
Anca Rath ◽  
Christoph Schoerner ◽  
Renate Meyer ◽  
Thomas Bertsch ◽  
...  
Keyword(s):  
Immunosorbent Assay ◽  
Roc Curve ◽  
Point Of Care ◽  
Lateral Flow ◽  
Lyme Neuroborreliosis ◽  
Diagnostic Tools ◽  
Lateral Flow Immunoassay ◽  
Enzyme Linked Immunosorbent Assay ◽  
European Federation ◽  
Point Of Care Test

ABSTRACT Diagnosis of Lyme neuroborreliosis (LNB) is challenging, as long as Borrelia-specific intrathecal antibodies are not yet detectable. The chemokine CXCL13 is elevated in the cerebrospinal fluid (CSF) of LNB patients. Here, we compared the performances of the Euroimmun CXCL13 enzyme-linked immunosorbent assay (CXCL13 ELISA) and the ReaScan CXCL13 lateral flow immunoassay (CXCL13 LFA), a rapid point-of-care test, to support the diagnosis of LNB. In a dual-center case-control study, CSF samples from 90 patients (34 with definite LNB, 10 with possible LNB, and 46 with other central nervous system [CNS] diseases [non-LNB group]) were analyzed with the CXCL13 ELISA and the CXCL13 LFA. Classification of patients followed the European Federation of Neurological Societies (EFNS) guidelines on LNB. The CXCL13 ELISA detected elevated CXCL13 levels in all patients with definite LNB (median, 1,409 pg/ml) compared to the non-LNB controls (median, 20.7 pg/ml; P < 0.0001), with a sensitivity of 100% and a specificity of 84.8% (cutoff value, 78.6 pg/ml; area under the receiver operating characteristic [ROC] curve, 0.93). Similarly, the CXCL13 LFA yielded elevated CXCL13 levels in 31 patients with definite LNB (median arbitrary value, 223.5) compared to the non-LNB control patients (median arbitrary value, 0; P < 0.0001) and had a sensitivity and specificity of 91.2% and 93.5%, respectively (cutoff arbitrary value, 22.5; area under the ROC curve, 0.94). The correlation between the CXCL13 levels obtained by ELISA and LFA was strong (Spearman correlation coefficient r = 0.89; P < 0.0001). The CXCL13 ELISA and the CXCL13 LFA are comparable diagnostic tools for the detection of CXCL13 in the CSF of patients with definite LNB. The advantage of the CXCL13 LFA is the shorter time to result.

scholarly journals Development of a multiplex and sensitive lateral flow immunoassay for the diagnosis of periprosthetic joint infection

2019 ◽  
Vol 9 (1) ◽  
Author(s):  
Tsung-Ting Tsai ◽  
Tse-Hao Huang ◽  
Natalie Yi-Ju Ho ◽  
Yu-Pei Chen ◽  
Chung-An Chen ◽  
...  
Keyword(s):  
Synovial Fluid ◽  
Clinical Diagnosis ◽  
Periprosthetic Joint Infection ◽  
Diagnostic Tests ◽  
Joint Infection ◽  
Lateral Flow ◽  
Rapid Response ◽  
Detection Sensitivity ◽  
Lateral Flow Immunoassay ◽  
Enzyme Linked Immunosorbent Assay

Abstract The diagnosis of periprosthetic joint infection (PJI) remains a challenge. However, recent studies showed that synovial fluid biomarkers have demonstrated greater diagnostic accuracy than the currently used PJI diagnostic tests. In many diagnostic tests, combining several biomarkers into panels is critical for improving diagnostic efficiency, enhancing the diagnostic precision for specific diseases, and reducing cost. In this study, we prove that combining alpha-defensin and C-reactive protein (CRP) as biomarkers possesses the potential to provide accurate PJI diagnosis. To further verify the result, we developed a multi-target lateral flow immunoassay strip (msLFIA) with staking pad design to obtain on-site rapid response for clinical diagnosis of PJI. A total of 10 synovial fluid samples were tested using the msLFIA, and the results showed that the combined measurements of synovial fluid alpha-defensin and CRP levels were consistent with those obtained from a commercial enzyme-linked immunosorbent assay kit. In addition, we developed a multi-target lateral flow immunoassay strip (msLFIA) with staking pad design to obtain on-site rapid response for clinical diagnosis of PJI, which the multi-target design is used to increase specificity and the stacking pad design is to enhance detection sensitivity. As a result, the turnaround time of the highly sensitive test can be limited from several hours to 20 min. We expect that the developed msLFIA possesses the potential for routine monitoring of PJI as a convenient, low-cost, rapid and easy to use detection device for PJI.

scholarly journals Lateral Flow Immunoassay with Quantum-Dot-Embedded Silica Nanoparticles for Prostate-Specific Antigen Detection

Nanomaterials ◽  
2021 ◽  
Vol 12 (1) ◽  
pp. 33
Author(s):  
Sungje Bock ◽  
Hyung-Mo Kim ◽  
Jaehi Kim ◽  
Jaehyun An ◽  
Yun-Sik Choi ◽  
...  
Keyword(s):  
Prostate Cancer ◽  
Silica Nanoparticles ◽  
Prostate Specific Antigen ◽  
False Negative ◽  
Specific Antigen ◽  
Calf Serum ◽  
High Sensitivity ◽  
Lateral Flow ◽  
Detection Sensitivity ◽  
Lateral Flow Immunoassay

Prostate cancer can be detected early by testing the presence of prostate-specific antigen (PSA) in the blood. Lateral flow immunoassay (LFIA) has been used because it is cost effective and easy to use and also has a rapid sample-to-answer process. Quantum dots (QDs) with very bright fluorescence have been previously used to improve the detection sensitivity of LFIAs. In the current study, a highly sensitive LFIA kit was devised using QD-embedded silica nanoparticles. In the present study, only a smartphone and a computer software program, ImageJ, were used, because the developed system had high sensitivity by using very bright nanoprobes. The limit of PSA detection of the developed LFIA system was 0.138 ng/mL. The area under the curve of this system was calculated as 0.852. The system did not show any false-negative result when 47 human serum samples were analyzed; it only detected PSA and did not detect alpha-fetoprotein and newborn calf serum in the samples. Additionally, fluorescence was maintained on the strip for 10 d after the test. With its high sensitivity and convenience, the devised LFIA kit can be used for the diagnosis of prostate cancer.

scholarly journals Serologic SARS-CoV-2 testing in healthcare workers with positive RT-PCR test or Covid-19 related symptoms

2020 ◽  
Author(s):  
Giovanni Visci ◽  
Vittorio Lodi ◽  
Roberta Bonfiglioli ◽  
Tiziana Lazzarotto ◽  
Francesco S. Violante ◽  
...  
Keyword(s):  
Healthcare Workers ◽  
Lateral Flow ◽  
Public Hospitals ◽  
Lateral Flow Immunoassay ◽  
Enzyme Linked Immunosorbent Assay ◽  
Limited Information ◽  
Rt Pcr ◽  
Positive Serology ◽  
Serologic Response ◽  
Pcr Test

AbstractBackgroundLimited information is available on prevalence and determinants of serologic response to SARS-CoV-2 infection among healthcare workers (HCWs).MethodsWe analyzed the results of serologic testing with chemiluminescence immunoassay analyzer (CLIA), lateral flow immunoassay (LFIA) and enzyme-linked immunosorbent assay (ELISA) test among 544 HCWs with at least one positive RT-PCR test and 157 HCWs with Covid-19 related symptoms without a positive RT-PCR test from public hospitals in Bologna, Northern Italy. Tests were performed between March and August 2020. We fitted multivariate logistic regression models to identify determinants of positive serology.ResultsThe sensitivity of SARS-CoV-2 was 75.2% (LFIA) and 90.6% (CLIA). No differences in seropositivity were observed by sex, while older HCWs had higher positivity than other groups, and nurses had higher positivity compared to physicians, but not other HCWs. An estimated 73.4% of HCWs with Covid-19 symptoms without RT-PCR test were not infected with SARS-CoV-2.ConclusionsOur study provides the best available data on sensitivity of serologic tests and on determinants of serologic response among HCWs positive for SARS-CoV-2, and provide evidence on the low specificity of Covid-19 related symptoms to identify infected HCWs.SummaryThe sensitivity of SARS-CoV-2 lateral flow immunoassay serology in healthcare workers (HCWs) was 75.2%. Older HCWs and nurses had higher positivity than other groups. An estimated 73.4% of HCWs with Covid-19 symptoms without RT-PCR test were not infected with SARS-CoV-2.

Evaluation of Detection Methods for Screening Meat and Poultry Products for the Presence of Foodborne Pathogens

2005 ◽  
Vol 68 (12) ◽  
pp. 2637-2647 ◽  
Author(s):  
VALERIE M. BOHAYCHUK ◽  
GARY E. GENSLER ◽  
ROBIN K. KING ◽  
JOHN T. WU ◽  
LYNN M. McMULLEN
Keyword(s):  
Foodborne Pathogens ◽  
Culture Method ◽  
Lateral Flow ◽  
Detection Methods ◽  
Enzyme Linked Immunosorbent Assay ◽  
Processed Meat ◽  
Culture Methods ◽  
E Coli ◽  
Poultry Products ◽  
Conventional Culture

Rapid and molecular technologies such as enzyme-linked immunosorbent assay (ELISA), PCR, and lateral flow immunoprecipitation can reduce the time and labor involved in screening food products for the presence of pathogens. These technologies were compared with conventional culture methodology for the detection of Salmonella, Campylobacter, Listeria, and Escherichia coli O157:H7 inoculated in raw and processed meat and poultry products. Recommended protocols were modified so that the same enrichment broths used in the culture methods were also used in the ELISA, PCR, and lateral flow immunoprecipitation assays. The percent agreement between the rapid technologies and culture methods ranged from 80 to 100% depending on the pathogen detected and the method used. ELISA, PCR, and lateral flow immunoprecipitation all performed well, with no statistical difference, compared with the culture method for the detection of E. coli O157:H7. ELISA performed better for the detection of Salmonella, with sensitivity and specificity rates of 100%. PCR performed better for the detection of Campylobacter jejuni, with 100% agreement to the culture method. PCR was highly sensitive for the detection of all the foodborne pathogens tested except Listeria monocytogenes. Although the lateral flow immunoprecipitation tests were statistically different from the culture methods for Salmonella and Listeria because of false-positive results, the tests did not produce any false negatives, indicating that this method would be suitable for screening meat and poultry products for these pathogens.

scholarly journals Development of a Europium Nanoparticles Lateral Flow Immunoassay for NGAL Detection in Urine and Diagnosis of Acute Kidney Injury

2021 ◽  
Author(s):  
Moli Yin ◽  
Yuanwang Nie ◽  
Hao Liu ◽  
Lei Liu ◽  
Lu Tang ◽  
...  
Keyword(s):  
Acute Kidney Injury ◽  
Kidney Injury ◽  
Lateral Flow ◽  
Adverse Outcomes ◽  
Detection Methods ◽  
Lateral Flow Immunoassay ◽  
Quantitative Detection ◽  
Detection Range ◽  
Neutrophil Gelatinase Associated Lipocalin ◽  
Europium Nanoparticles

Abstract Background:AKI is related to severe adverse outcomes and mortality with Coronavirus Infection Disease 2019 (COVID-19) patients, that early diagnosed and intervened is imperative. Neutrophil gelatinase-associated lipocalin (NGAL) is one of the most promising biomarkers for detection of acute kidney injury (AKI), but current detection methods are inadequacy, so more rapid, convenient and accuracy methods are needed to detect NGAL for early diagnosis of AKI. Herein, we established a rapid, reliable and accuracy lateral flow immunoassay based on europium nanoparticles (Eu-NPS-LFIA) for the detection of NGAL in human urine specimens. Methods:A double-antibody sandwich immunofluorescent assay using europium doped nanoparticles was employed and the NGAL monoclonal antibodies conjugate as labels were generated by optimizing electric fusion parameters. Eighty-three urine samples were used to evaluate the clinical application efficiency of this method. Results:The quantitative detection range of NGAL in AKI was 1-3000 ng/mL, and the detection sensitization was 0.36 ng/mL. The CV of intra-assay and inter-assay were 2.57%-4.98% and 4.11%-7.83%, respectively. Meanwhile, the correlation coefficient between Eu-NPS-LFIA and ARCHITECT analyzer was significant (R2=0.9829, n=83, p<0.01). Conclusions:Thus, a faster and easier operation quantitative assay of NGAL for AKI has been established, which is very important and meaningful to diagnose the early AKI, suggesting that the assay can provide an early warning of final outcome of disease.

scholarly journals A Rapid and Sensitive Fluorescent Microsphere-Based Lateral Flow Immunoassay for Determination of Aflatoxin B1 in Distillers’ Grains

Foods ◽  
2021 ◽  
Vol 10 (9) ◽  
pp. 2109
Author(s):  
Zifei Wang ◽  
Pengjie Luo ◽  
Baodong Zheng
Keyword(s):  
Aflatoxin B1 ◽  
High Performance ◽  
Animal Feed ◽  
Limit Of Detection ◽  
Lateral Flow ◽  
Distillers Grains ◽  
Detection Methods ◽  
Lateral Flow Immunoassay ◽  
Quantitative Detection ◽  
Fluorescent Microsphere

Aflatoxin B1 (AFB1) is a toxic compound naturally produced by the genera Aspergillus. Distillers’ grains can be used as animal feed since they have high content of crude protein and other nutrients. However, they are easily contaminated by mycotoxins, and currently there are no rapid detection methods for AFB1 in distillers’ grains. In this study, a lateral flow immunoassay (LFIA) based on red fluorescent microsphere (FM), is developed for quantitative detection of AFB1 in distillers’ grains. The whole test can be completed within 15 min, with the cut-off value being 25.0 μg/kg, and the quantitative limit of detection (qLOD) being 3.4 μg/kg. This method represents satisfactory recoveries of 95.2–113.0%, and the coefficients of variation (CVs) are less than 7.0%. Furthermore, this technique is successfully used to analyze AFB1 in real samples, and the results indicates good consistency with that of high-performance liquid chromatography (HPLC). The correlation coefficient is found to be greater than 0.99. The proposed test strip facilitates on-site, cost-effective, and sensitive monitoring of AFB1 in distillers’ grains.

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