Development of a nanoparticle-assisted PCR assay for detection of porcine epidemic diarrhea virus

Yuan, Wanzhe; Li, Yanan; Li, Peng; Song, Qinye; Liu, Limin; Sun, Jianhe

doi:10.1016/j.jviromet.2015.04.008

Cited by 32 publications

(20 citation statements)

References 17 publications

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“…The traditional identification methods may not provide accurate distinction of the attenuated vaccine strain for the field strain. The nanoparticle-assisted RT-PCR assay has been used for rapid and accurate detection of other pig viruses [24][25][26]. The present study demonstrated that an effective nanoparticle-assisted RT-PCR assay targeting the ORF3 gene of PEDV was able to distinguish field strains from attenuated vaccine strains.…”

Section: Discussionmentioning

confidence: 65%

Establishment of a nanoparticle-assisted RT-PCR assay to distinguish field strains and attenuated strains of porcine epidemic diarrhea virus

Zhu

Wang²,

Cui

et al. 2016

Arch Virol

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Porcine epidemic diarrhea virus (PEDV) can cause serious disease and even death in neonatal piglets, resulting in serious damage to the swine industry worldwide. Open reading frame 3 (ORF3) is the only accessory gene in the PEDV genome. Previous studies have indicated that PEDV vaccine strains have a partial deletion in ORF3. In this study, a nanoparticle-assisted polymerase chain reaction (nanoparticle-assisted RT-PCR) assay targeting the ORF3 of PEDV was developed to distinguish PEDV field strains from attenuated strains by using a specific pair of primers. The PCR products of field strains and attenuated strains were 264 bp and 215 bp in length, respectively. The sensitivity and specificity of this assay were also assessed. The nanoparticle-assisted RT-PCR assay was 10-100 times more sensitive than the conventional RT-PCR assay, with no cross-reactions when amplifying porcine pseudorabies virus (PRV), porcine circovirus type 2 (PCV2), classical swine fever virus (CSFV), porcine parvovirus (PPV), porcine reproductive and respiratory syndrome virus (PRRSV), porcine rotavirus (RV), and porcine transmissible gastroenteritis virus (TGEV). The nanoparticle-assisted RT-PCR assay we describe here can be used to distinguish field strains from vaccine strains of PEDV, and it shows promise for reducing economic loss due to PEDV infection.

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Section: Discussionmentioning

confidence: 65%

Establishment of a nanoparticle-assisted RT-PCR assay to distinguish field strains and attenuated strains of porcine epidemic diarrhea virus

Zhu

Wang²,

Cui

et al. 2016

Arch Virol

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“…To date, several approaches have been developed for the laboratory diagnosis of PEDV, including immunohistochemistry [46], enzymelinked immunosorbent assay (ELISA) [47], RT-PCR [48], reverse transcription loop-mediated isothermal amplification (RT-LAMP) [49], a nanoparticle-assisted PCR assay [50,51], and real-time RT-PCR [35,36]. Similarly, immunohistochemical detection, PCR, and qPCR have all been developed for the detection of PCV3 [20,33,34,37].…”

Section: Discussionmentioning

confidence: 99%

Development of a SYBR green I-based duplex real-time fluorescence quantitative PCR assay for the simultaneous detection of porcine epidemic diarrhea virus and porcine circovirus 3

Han

Zheng

Zhao

et al. 2019

Molecular and Cellular Probes

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A B S T R A C TThe development of a rapid, specific, and sensitive SYBR Green I-based duplex real-time quantitative PCR assay is described for the simultaneous detection of porcine epidemic diarrhea virus (PEDV) and porcine circovirus type 3 (PCV3). The assay specifically detected PEDV and PCV3, with no fluorescence detected for other nontargeted pig pathogens. The assay showed a good linear relationship, and the limits of detection for this assay were 34.6 copies/μL and 61.2 copies/μL for PEDV and PCV3, respectively. The assay exhibited high repeatability and reproducibility, with intra-assay and inter-assay variation coefficients less than 2.0%. A clinical evaluation using intestinal tissue and fecal samples from piglets suffering from diarrhea at different pig farms in China revealed that the singular infection rates of PEDV and PCV3 were 43.94% (29/66) and 16.67% (11/66), respectively, while the co-infection rate of PCV3 with PEDV was 27.27% (18/66). The results indicate this assay is a rapid and reliable diagnostic tool for PEDV and PCV3 monitoring and surveillance in the field, and provides technical support for the quantitative detection of clinical samples infected or co-infected with PEDV and PCV3.

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“…Currently, there are several ways to detect PEDV, such as molecular biological detection methods including nucleic acid probes, polymerase chain reaction (PCR), and loop-mediated isothermal amplification among others. These methods have both high precision and specificity [9][10][11][12][13][14]. However, they require precise experimental instruments and trained experimenters, and have certain restrictions on promoting grassroots production.…”

Section: Introductionmentioning

confidence: 99%

EuNPs-mAb fluorescent probe based immunochromatographic strip for rapid and sensitive detection of porcine epidemic diarrhea virus

Jin

Zou

et al. 2020

Talanta

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A B S T R A C TPorcine epidemic diarrhea (PED), induced by porcine epidemic diarrhea virus (PEDV) causes acute diarrhea, vomiting, dehydration and high mortality in neonatal piglets, resulting in significant economic losses in the pig industries. In this study, an immunochromatographic assay (ICA) based on a EuNPs-mAb fluorescent probe was developed and optimized for rapid detection of PEDV. The limit of detection (LOD) of the ICA was 0.218 μg/mL (2.725 × 10 3 TCID 50 /mL) and its linear detection range was 0.03125-8 μg/mL (3.91 × 10 2 -10 5 TCID 50 /mL). The ICA was also validated for the detection of PEDV in swine stool samples. 60 swine stool samples from southern China were analyzed by the ICA and RT-PCR, and the results showed that the coincidence rate of the ICA to RT-PCR was 86.67%, which was significantly higher than that of AuNPs based ICA. The ICA is sensitive and specific and can achieve on-site rapid detection of swine stool samples. Therefore, the ICA has a great potential for PED diagnosis and prevention.

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Development of a nanoparticle-assisted PCR assay for detection of porcine epidemic diarrhea virus

Cited by 32 publications

References 17 publications

Establishment of a nanoparticle-assisted RT-PCR assay to distinguish field strains and attenuated strains of porcine epidemic diarrhea virus

Establishment of a nanoparticle-assisted RT-PCR assay to distinguish field strains and attenuated strains of porcine epidemic diarrhea virus

Development of a SYBR green I-based duplex real-time fluorescence quantitative PCR assay for the simultaneous detection of porcine epidemic diarrhea virus and porcine circovirus 3

EuNPs-mAb fluorescent probe based immunochromatographic strip for rapid and sensitive detection of porcine epidemic diarrhea virus

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