Development of a duplex SYBR GreenⅠ based real-time PCR assay for detection of porcine epidemic diarrhea virus and porcine bocavirus3/4/5

Zheng, Lan Lan; Cui, Jian; Han, Hao Ying; Hou, Hua Lin; Wang, Leyi; Liu, Fang; Chen, Hong Ying

doi:10.1016/j.mcp.2020.101544

Cited by 10 publications

(4 citation statements)

References 23 publications

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“…quantification of PBoV and pseudorabies virus (Luo et al 2015). SYBR Green-based duplex real-time PCR has been used for detection of porcine epidemic diarrhea virus (PEDV) as well as PBoV genotypes (3,4,5), with detection limits of 10 copies/lL for both viruses (Zheng et al 2020). Sequence-independent single primer amplification (SISPA) is a primer-initiated method that uses a single primer to amplify nucleic acids of an unknown sequence by sequenceindependent PCR.…”

Section: Detection Methods Of Pbovmentioning

confidence: 99%

“…However, the co-occurrence of this virus with circulating viruses has been reported since its discovery (Table 1) (Zhou et al 2017;. The most common circulating viruses are PEDV (Zhang et al 2013;Zheng et al 2020), GARV (Zhang et al 2013) (Zhang et al 2011;Zhou et al 2018), porcine parvovirus , porcine pseudorabies virus (Luo et al 2015) and porcine kobuvirus (Zhang et al 2013). Clinically, the incidence of coinfection of PBoV with PCV-2 has been reported to be 83.8%, suggesting that PCV-2 enhances the infectivity of PBoV ).…”

Section: Coinfection Of Pbov With Typical Swine Enteric Virusesmentioning

confidence: 99%

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Porcine Bocavirus: A 10-Year History since Its Discovery

Aryal

Liu

2021

Virol. Sin.

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Porcine bocavirus (PBoV) is a single-stranded DNA virus, belongs to the genus Bocaparvovirus of family Parvoviridae. It was discovered along with porcine circovirus 2 (PCV 2) and torque tenovirus (TTV) in the lymph nodes of pigs suffering from postweaning multisystemic wasting syndrome (PMWS) in Sweden in 2009. PBoV has been reported throughout the world, mostly in weaning piglets, and has a broad range of tissue tropism. Since PBoV is prevalent in healthy as well as clinically infected pigs and is mostly associated with coinfection with other viruses, the pathogenic nature of PBoV is still unclear. Currently, there are no cell lines available for the study of PBoV, and animal model experiments have not been described. This review summarizes the current state of knowledge about PBoV, including the epidemiology, evolution analysis, detection methods, pathogenesis and public health concerns.

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Section: Detection Methods Of Pbovmentioning

confidence: 99%

Section: Coinfection Of Pbov With Typical Swine Enteric Virusesmentioning

confidence: 99%

Porcine Bocavirus: A 10-Year History since Its Discovery

Aryal

Liu

2021

Virol. Sin.

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“…In addition to PCR, quantitative real-time PCR (qRT-PCR), a more sensitive and rapid assay, had also been widely used. Specific primers targeting the N gene sequence of PEDV have been used for real-time PCR assays based using SYBR Green І detection chemistry ( Zheng et al, 2020 ). In general, the specificity and sensitivity of TaqMan probe real-time PCR methods are considered better than intercalating dye-based real-time PCR.…”

Section: Introductionmentioning

confidence: 99%

Clustered Regularly Interspaced Short Palindromic Repeat/Cas12a Mediated Multiplexable and Portable Detection Platform for GII Genotype Porcine Epidemic Diarrhoea Virus Rapid Diagnosis

et al. 2022

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Porcine epidemic diarrhoea virus (PEDV) is a member of the genus Alphacoronavirus in the family Coronaviridae. It causes acute watery diarrhoea and vomiting in piglets with high a mortality rate. Currently, the GII genotype, PEDV, possesses a high separation rate in wild strains and is usually reported in immunity failure cases, which indicates a need for a portable and sensitive detection method. Here, reverse transcription–recombinase aided amplification (RT-RAA) was combined with the Clustered Regularly Interspaced Short Palindromic Repeat (CRISPR)/Cas12a system to establish a multiplexable, rapid and portable detection platform for PEDV. The CRISPR RNA (crRNA) against Spike (S) gene of GII PEDV specifically were added into the protocol. This system is suitable for different experimental conditions, including ultra-sensitive fluorescence, visual, UV light, or flow strip detection. Moreover, it exhibits high sensitivity and specificity and can detect at least 100 copies of the target gene in each reaction. The CRISPR/Cas12a detection platform requires less time and represents a rapid, reliable and practical tool for the rapid diagnosis of GII genotype PEDV.

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“…(A) Single LAMP–LFD sensitivity testing, (B) duplex LAMP–LFD sensitivity testing; Table S1: Sequences of qPCR primers and probes for detecting PEDV, PoRV and PBoV; Table S2: Informations of 125 animal feces samples; Table S3: Comparison of molecular methods for the detection of porcine diarrhea virus. References [ 25 , 50 , 54 , 55 , 56 , 57 , 58 , 59 , 60 , 61 , 62 , 63 , 64 , 65 , 66 , 67 , 68 , 69 , 70 , 71 ] are cited in the supplementary materials.…”

mentioning

confidence: 99%

Triplex-Loop-Mediated Isothermal Amplification Combined with a Lateral Flow Immunoassay for the Simultaneous Detection of Three Pathogens of Porcine Viral Diarrhea Syndrome in Swine

Yang

Li³

et al. 2023

Animals

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Porcine epidemic diarrhea virus (PEDV), porcine bocavirus (PBoV), and porcine rotavirus (PoRV) are associated with porcine viral diarrhea. In this study, triplex loop-mediated isothermal amplification (LAMP) combined with a lateral flow dipstick (LFD) was established for the simultaneous detection of PEDV, PoRV, and PBoV. The PEDV-gp6, PoRV-vp6, and PBoV-vp1 genes were selected to design LAMP primers. The amplification could be carried out at 64 °C using a miniature metal bath within 30 min. The triplex LAMP–LFD assay exhibited no cross-reactions with other porcine pathogens. The limits of detection (LODs) of PEDV, PoRV, and PBoV were 2.40 × 101 copies/μL, 2.89 × 101 copies/μL, and 2.52 × 101 copies/μL, respectively. The consistency between rt-qPCR and the triplex LAMP–LFD was over 99% in field samples testing. In general, the triplex LAMP–LFD assay was suitable for the rapid and simultaneous detection of the three viruses in the field.

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Development of a duplex SYBR GreenⅠ based real-time PCR assay for detection of porcine epidemic diarrhea virus and porcine bocavirus3/4/5

Cited by 10 publications

References 23 publications

Porcine Bocavirus: A 10-Year History since Its Discovery

Porcine Bocavirus: A 10-Year History since Its Discovery

Clustered Regularly Interspaced Short Palindromic Repeat/Cas12a Mediated Multiplexable and Portable Detection Platform for GII Genotype Porcine Epidemic Diarrhoea Virus Rapid Diagnosis

Triplex-Loop-Mediated Isothermal Amplification Combined with a Lateral Flow Immunoassay for the Simultaneous Detection of Three Pathogens of Porcine Viral Diarrhea Syndrome in Swine

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