Development of a rapid and sensitive europium (III) chelate microparticle-based lateral flow test strip for the detection and epidemiological surveillance of porcine epidemic diarrhea virus

Abstract

Porcine epidemic diarrhea virus (PEDV), a member of the genus Alphacoronavirus, is the predominant cause of severe enteropathogenic diarrhea in swine. A simple, rapid, specific, and sensitive method is critical for monitoring PEDV on pig farms. In this study, a simple and rapid lateral flow immunoassay detection system that integrates europium (Eu) (III) chelate microparticles was developed to identify PEDV in fecal swabs. This newly developed diagnostic sandwich immunoassay utilizes lateral flow test strips (LFTSs). The fluorescence peak heights of the test line (HT) and the control line (HC) were measured using a fluorescence strip reader, and the HT/HC ratio was used for quantitation. The limit of detection of PEDV with this LFTS was ??ten times the median tissue culture infectious dose (TCID 50 ) per mL??. Fecal swab samples were used to determine the cutoff value. Field samples, various PEDV strains and other viruses were used to determine the sensitivity and specificity of the Eu (III) chelate microparticle-based LFTSs, which were 97.8% and 100%, respectively, with a cutoff value of 0.05, as compared with reverse transcription polymerase chain reaction (RT-PCR). In samples from piglets experimentally infected with PEDV, the results were in high agreement with those obtained by RT-PCR. Epidemiological surveillance of PEDV using the LFTSs ??in areas threatened by African swine fever virus?? suggested that the PEDV positive rate on pig farms had significantly decreased, mainly due to the implementation of strict biosecurity measures. The results indicate that the Eu (III) chelate microparticle-based LFTS system is a rapid, sensitive, and reliable method for the identification of PEDV, indicating its suitability for epidemiological surveillance of PEDV infection.Publisher's Note Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Point-of-Care Tests for Rapid Detection of Porcine Epidemic Diarrhea Virus: A Systematic Review and Meta-Analysis

“…Further influential analysis using a bivariate box plot and Cook’s distance revealed an outlier study for ICA-based POCTs and four outliers for NAIA-based POCTs ( Supplementary File Figures S3 and S4 ). After removing these outliers [ 21 , 35 , 36 , 37 , 41 ], the pooled sensitivities of ICA- and NAIA-based POCTs for detecting PEDV showed significantly statistical differences comparing with those of the corresponding overall studies (0.94, 95% CI 0.92–0.97 vs. 0.95, 95% CI 0.93–0.97, p = 0.00 for ICA-based POCTs; 0.98, 95% CI 0.96–1.00 vs. 0.97, 95% CI 0.95–0.99 for NAIA-based POCTs, p = 0.01), indicating that these outliers may also have contributed to heterogeneity among the studies.…”

“…In addition, some ICA-based tests provide qualitative information on the presence or absence of PEDV with the recent development of several instruments developed to measure the intensity of the test line for a more quantitative judgment through attaching to optic or electrochemical detectors. In this systematic review and meta-analysis, we identified 11 articles of ICA-based POCTs published since 2015, and 2 of them used portable strip readers to analyze the detection results of ICA [ 19 , 21 ]. Moreover, a smartphone camera and immunofluorescent analyzer were also applied to quantify the results of ICA for detecting PEDV [ 18 , 22 ].…”

Evolution, Interspecies Transmission, and Zoonotic Significance of Animal Coronaviruses

“…The diagnosis for a newly identified pathogen, porcine delta CoV, was made possible by developing a cost-effective fluorescent microsphere immunoassay as used for PEDV ( 351 ) that could detect antibodies against multiple target antigens of porcine delta CoV for better efficiency and sero-surveillance on a herd level ( 352 ). Recently, a europium (III) chelate microparticle-based lateral flow test strip was developed for identification and epidemiological surveillance of PEDV with high reliability and sensitivity ( 353 ). A novel multiplex PCR-electronic microarray assay for rapid and comprehensive detection of bovine respiratory and enteric pathogens, including BCoV, was also developed ( 354 ).…”

COVID-19: A Mechanistic Approach to Understand the Detrimental Consequences for Vaccine Development and Perspectives

“…2(c,d) corroborates the central dogma and internally oriented genetic material followed by the fragmentation of template copies of ssRNA through the replication. However, this COVID-19 has been referred as SARS-CoV2 based on its detrimental consequences and symptoms [12]. It has been observed that the protein sequence of COVID-19 and SARS-CoV2 are paralogous which are the ancestral gene followed by the duplication for further evolution.…”

“…It is the very specific characteristic property of the COVID-19 virus which reverse invades against the immune system (antibodies). At this stage natural killer (T-cells) will also become neutral and cannot show much activity on virus [11][12][13][14]. Further, Fig.…”