A novel diagnostic approach to detecting porcine epidemic diarrhea virus: The lateral immunochromatography assay

Kim, Yong Kwan; Lim, Suk-Kyung; Cho, In‐Soo; Cheong, Kwang-Myun; Lee, Eun-Jeong; Lee, Sang‐Oh; Kim, Joon-Bae; Kim, Jung-Hwa; Jeong, Dong Seop; An, Byung-Hyun; An, Dong-Jun

doi:10.1016/j.jviromet.2015.08.024

Cited by 20 publications

(18 citation statements)

References 38 publications

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“…Therefore, it is necessary to definitely identify the causative pathogen using confirmatory laboratory tests, which is essential for timely clinical decision-making and management of epidemics of PED [13]. Commonly used laboratory diagnostic methods of PEDV include antigen enzyme-linked immunosorbent assay (ELISA), immunochromatography assay (IC), reverse transcriptase polymerase chain reaction (RT-PCR), reverse transcription loop-mediated isothermal amplification (RT-LAMP), Taq-Man-based real-time RT-PCR and nanoparticle-assisted PCR assay and so forth [14][15][16][17][18][19][20]. Currently, ELISA and IC have been widely applied to detect PEDV in large-scale blood or feces samples.…”

Section: Introductionmentioning

confidence: 99%

Ultrasensitive Detection of Porcine Epidemic Diarrhea Virus from Fecal Samples Using Functionalized Nanoparticles

Xing

Guan

et al. 2016

PLoS ONE

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Porcine epidemic diarrhea virus (PEDV) is the main causative agent of porcine diarrhea, which has resulted in devastating damage to swine industry and become a perplexed global problem. PEDV infection causes lesions and clinical symptoms, and infected pigs often succumb to severe dehydration. If there is not a timely and effective method to control its infection, PEDV will spread rapidly across the whole swine farm. Therefore, preclinical identification of PEDV is of great significance for preventing the outbreak and spread of this disease. In this study, a functionalized nanoparticles-based PCR method (UNDP-PCR) specific for PEDV was developed through systematic optimization of functionalized magnetic beads and gold nanoparticles which were further used to specifically enrich viral RNA from the lysate of PEDV stool samples, forming a MMPs-RNA-AuNPs complex. Then, oligonucleotides specific for PEDV coated on AuNPs were eluted from the complex and were further amplified and characterized by PCR. The detection limitation of the established UNDP-PCR method for PEDV was 25 copies in per gram PEDV stool samples, which is 400-fold more sensitive than conventional RT-PCR for stool samples. The UNDP-PCR for PEDV exhibited reliable reproducibility and high specificity, no cross-reaction was observed with other porcine viruses. In 153 preclinical fecal samples, the positive detection rate of UNDP-PCR specific for PEDV (30.72%) was much higher than that of conventional RT-PCR (5.88%) and SYBR Green real-time RT-PCR. In a word, this study provided a RNA extraction and transcription free, rapid and economical method for preclinical PEDV infection, which showed higher sensitivity, specificity and reproducibility, and exhibited application potency for evaluating viral loads of preclinical samples.

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Section: Introductionmentioning

confidence: 99%

Ultrasensitive Detection of Porcine Epidemic Diarrhea Virus from Fecal Samples Using Functionalized Nanoparticles

Xing

Guan

et al. 2016

PLoS ONE

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“…Therefore, it is of great importance to develop differential laboratory tests [9,10]. To date, several methods including RT-PCR [11], separation identification [12], serological method [13], enzyme-linked immunosorbent assay (ELISA) [14] and colloidal gold method [15][16][17] have been used to detect the PEDV antigen. Although RT-PCR serves as a good standard due to its high sensitivity and accuracy, this method relies heavily on sophisticated equipment and expensive apparatus.…”

Section: Introductionmentioning

confidence: 99%

A new immunochromatographic assay for on-site detection of porcine epidemic diarrhea virus based on monoclonal antibodies prepared by using cell surface fluorescence immunosorbent assay

Bian

Jia

et al. 2019

BMC Vet Res

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BackgroundPorcine epidemic diarrhea virus (PEDV) is a highly effective pathogen that can cause death of new-born piglet, resulting in big economical loss in pig farming industry. For rapid detection of PEDV, a new immunochromatographic assay (ICA) based on monoclonal antibodies (mAbs) was developed in this study.ResultsThe mAbs were prepared by using PEDV positive hybridoma cells that were selected by using cell surface fluorescence immunosorbent assay (CSFIA). Fourteen mAbs against PEDV strain isolated from south of China were prepared. The optimal mAb 4A11 was coated on NC membrane as the capturing reagent and the mAb A11H7 was coupled to gold nanoparticles (AuNPs) as detection reagent for the new ICA. The new ICA was used to measure PEDV in phosphate buffer containing tween-20. Results indicated that the limit of detection (LOD) of the new ICA was 0.47 μg/mL (5.9 × 103 TCID50/mL) and the liner detection range of the ICA was 0.625–10 μg/mL (7.8 × 103–105 TCID50/mL). The specificity analysis results showed that this new ICA had no cross reaction in the presence of other porcine viruses. The ICA was also validated for the detection of PEDV in swine stool samples with little interference from swine stool. To compare its accuracy to other traditional detection methods, 27 swine stool samples from south of China were investigated with the new developed ICA, commercial strip and RT-PCR. Results showed that the new ICA was more comparable to RT-PCR than commercial test strip.ConclusionsA new ICA based on mAbs prepared by CSFIA was developed in this study. It was a sensitive, specific and rapid method that could be used for on-site detection of PEDV and therefore was useful for the diagnosis and prevention of PED.Electronic supplementary materialThe online version of this article (10.1186/s12917-019-1773-4) contains supplementary material, which is available to authorized users.

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“…The methods currently used to diagnose PED in clinical samples are mainly divided into two types: 1) immunological methods including immunochromatography and enzyme-linked immunosorbent assay (ELISA) [8,9], and 2) molecular biology assays including conventional reverse transcription-polymerase chain reaction (RT-PCR), reverse transcription quantitative PCR (RT-qPCR), multiplex PCR, and reverse transcription loop-mediated isothermal amplification (RT-LAMP) [10][11][12][13]. Although immunological methods are generally low cost and easy to perform, they have several disadvantages, including inconclusive results and the long time required to perform the assays.…”

Section: Introductionmentioning

confidence: 99%

Visual detection of porcine epidemic diarrhea virus using a novel reverse transcription polymerase spiral reaction method

Wang

et al. 2019

BMC Vet Res

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Background Porcine epidemic diarrhea virus (PEDV) is a major etiological agent of porcine epidemic diarrhea around the world. Point-of-care testing in the field is lacking owing to the requirement for a simple, robust field applicable test that does not require professional laboratory equipment. The aim of this study was to establish a novel reverse transcription polymerase spiral reaction (RT-PSR) assay for the rapid detection of porcine epidemic diarrhea virus (PEDV). For the assay, a specific RT-PSR primer pair was designed against a conserved region in PEDV ORF3. Results The RT-PSR was optimized, and PEDV could be detected after a 50 min incubation at 62 °C, in addition to the 15 min required for reverse transcription. No cross-reaction with other porcine infectious viruses was observed. This new method for PEDV detection was 10 times more sensitive than the conventional reverse transcription-polymerase chain reaction (RT-PCR) assay. The positive rates for 65 clinical samples using the new RT-PSR assay and the conventional RT-PCR assay were 58.46% (38/65) and 53.84% (35/65), respectively. In the RT-PSR assay, the addition of a mixture of dyes allowed a positive reaction to be directly observed by the naked eye. Conclusions These results indicate that this RT-PSR assay is capable of accurately detecting PEDV, and has the advantages of high specificity and sensitivity for the detection of PEDV.

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A novel diagnostic approach to detecting porcine epidemic diarrhea virus: The lateral immunochromatography assay

Cited by 20 publications

References 38 publications

Ultrasensitive Detection of Porcine Epidemic Diarrhea Virus from Fecal Samples Using Functionalized Nanoparticles

Ultrasensitive Detection of Porcine Epidemic Diarrhea Virus from Fecal Samples Using Functionalized Nanoparticles

A new immunochromatographic assay for on-site detection of porcine epidemic diarrhea virus based on monoclonal antibodies prepared by using cell surface fluorescence immunosorbent assay

Visual detection of porcine epidemic diarrhea virus using a novel reverse transcription polymerase spiral reaction method

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