Concurrent infections of pseudorabies virus and porcine bocavirus in China detected by duplex nanoPCR

Luo, Yakun; Liang, Lin; Zhou, Ling; Zhao, Kai; Cui, Shangjin

doi:10.1016/j.jviromet.2015.03.016

Cited by 14 publications

(10 citation statements)

References 17 publications

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“…The high co-infection rate of PCV2 and PBoV in this study demonstrated that the higher PBoV prevalence in samples from pigs with PWMS, which were similar to results reported previously (Blomstrom et al, 2010), suggested that PBoV might play a role in the development of PMWS. In addition, the co-infection of PBoV with other pathogens was common, such as PRRSV, CSFV (Blomstrom et al, 2010;Meng, 2012;McMenamy et al, 2013;Luo et al, 2015). Pfankuche et al (Pfankuche et al, 2016) reported that PBoV should be considered as a pathogen that triggers encephalomyelitis.…”

Section: Resultsmentioning

confidence: 99%

“…Co-infection of PBoV with other porcine viruses has also been reported, such as porcine epidemic diarrhea virus (PEDV), porcine circovirus type 2 (PCV2), pseudorabies virus (PRV), porcine reproductive and respiratory syndrome virus (PRRSV), porcine torque teno virus (PTTV) and classic swine fever virus (CSFV) (Blomstrom et al, 2010;McMenamy et al, 2013;Zhang et al, 2014;Huang et al, 2014;Zhou et al, 2014;Luo et al, 2015). PBoV was detected from livers of healthy pigs with prevalence of 6% in Brazil, and the co-infection of PBoV with torque teno sus virus 1(TTSuV1), TTSuVk2 and porcine parvovirus (PPV) was also found in these healthy pigs (Silva et al, 2020).…”

Section: Introductionmentioning

confidence: 99%

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Detection and genetic characteristic of porcine bocavirus in central China

Zheng¹,

Cui²,

Han³

et al. 2020

Preprint

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To investigate the epidemic profile and genetic diversity of porcine bocavirus (PBoV), 281 clinical samples including 236 intestinal tissue samples and 45 fecal samples were collected from diarrheal piglets in 37 different pig farms of central China, and two SYBR Green I-based quantitative PCR assays were developed to detect PBoV1/2 and PBoV3/4/5 respectively. The results showed the detection limits of two assays were 1.66 × 101 genome copies/μl of PBoV1/2 and 3.3 × 101 copies/μL of PBoV 3/4/5. 148 (52.67%) of the 281 clinical samples were positive for PBoV1/2, 117 (41.63%) were positive for PBoV3/4/5, 55 (19.57%) were positive for both PBoV1/2 and PBoV3/4/5, and 86.49% (32/37) of the pig farms were positive for PBoV. Subsequently, complete genomic sequences of two PBoV strains (designated CH/HNZM and PBoV-TY) from two different farms were sequenced. The phylogenetic analysis demonstrated that the two PBoV strains obtained in this study belonged to the PBoV2 group and had a close relationship with other 12 PBoV2 strains, but differed genetically from PBoV1, PBoV3/4/5 and 7 other bocaviruses. CH/HNZM and PBoV-TY were closely related to the PBoV strain GD18 (KJ755666) which may be derived from PBoV strains 0912/2012 (MH558677) and 57AT-HU (KF206160) through the recombination analysis. Compared with reference strain ZJD (HM053694)-China, a higher amino acid variation was found in the NS1 protein of CH/HNZM and PBoV-TY. These results extend our understanding of the molecular epidemiology and evolution of PBoV.

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Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Detection and genetic characteristic of porcine bocavirus in central China

Zheng¹,

Cui²,

Han³

et al. 2020

Preprint

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“…Since reagents are limited in the multiple PCR amplification system, each pair of primers compete for the reagents in the reaction. To enhance amplification, the volume and ratios of the three pairs of primers need to be optimized (Luo et al, 2015;Wang et al, 2015). The volumes of all the primers (10 µM) used ranged from 0.1 to 1.0 µL in increments of 0.1 µL.…”

Section: Optimization Of the Conditions For Dpo-nanopcr Assaymentioning

confidence: 99%

Simultaneous Detection of Bovine Rotavirus, Bovine Parvovirus, and Bovine Viral Diarrhea Virus Using a Gold Nanoparticle-Assisted PCR Assay With a Dual-Priming Oligonucleotide System

et al. 2019

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Bovine rotavirus (BRV), bovine parvovirus (BPV), and bovine viral diarrhea virus (BVDV) are the pathogens that cause diarrhea primarily in newborn calves. A mixed infection of BRV, BPV, and BVDV makes clinical diagnosis difficult. In this study, we designed dual-priming oligonucleotide (DPO) primers the VP6 gene of BRV, VP2 gene of BPV, and 5 UTR gene of BVDV and synthesized gold nanoparticles (GNPs) with an average diameter of 10 nm. We combined the DPOs with the GNPs to develop a DPO-nanoPCR assay for detecting BRV, BPV, and BVDV. The annealing temperature, primer concentration, and GNP concentration were optimized for this assay. Compared to a conventional PCR assay, the DPO-nanoPCR assay allowed the use of a wider range of annealing temperatures (41-65 • C) to effectively amplify target genes. PCR amplification was the most efficient at 56.2 • C using conventional primers. The optimal volume of all the primers (10 µM) was 1.0 µL. The optimal volume of GNPs (10 nM) for all the reactions was 0.5 µL.

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“…To date, pigs infected with PBoV have been reported in 20 provinces or regions in China with the prevalence between 7.3% and 64% (Wang et al, 2014;Zhang et al, 2015;Zhou et al, 2018). Co-infection of PBoV with other porcine viruses has also been reported, such as porcine epidemic diarrhea virus (PEDV), porcine circovirus type 2 (PCV2), pseudorabies virus (PRV), porcine reproductive and respiratory syndrome virus (PRRSV), porcine torque teno virus (PTTV) and classic swine fever virus (CSFV) (Blomstrom et al, 2010;McMenamy et al, 2013;Zhang et al, 2014;Huang et al, 2014;Zhou et al, 2014;Luo et al, 2015). In addition, PBoV has a significantly higher infection rate in diseased pigs than in healthy pigs, and the coinfection rate of PEDV and PBoV was higher in samples of diarrheal pigs than that of healthy pigs, suggesting that the PBoV might play an important role in causing diarrhea in piglets (Zhai et al, 2010).…”

Section: Introductionmentioning

confidence: 99%

Molecular detection, phylogenetic and evolutionary analysis of porcine bocavirus in central China

Zheng¹,

Cui²,

Qiao³

et al. 2020

Preprint

View full text Add to dashboard Cite

To investigate the phylogenetic and evolutionary analysis of porcine bocavirus (PBoV), 281 clinical samples including 236 intestinal tissue samples and 45 fecal samples were collected from diarrheal piglets in 27 different pig farms of central China, and SYBR Green I-based qPCR assays were developed to detect PBoVs. Among the 281 samples, 148 (52.67%) were positive for PBoV1/2, 117 (41.63%) were positive for PBoV3/4/5, and 55 (19.57%) were positive for both PBoV1/2 and PBoV3/4/5. Subsequently, two complete genome sequences of PBoV strains CH/HNZM (accession numbers KX017193) and PBoV-TY (accession numbers MH454686) were cloned in this study. A multiple genomic sequence alignment showed that the pairwise similarity of the two PBoV strains was 94.8%, and they had 44.5%-95.8% genomic nucleotide sequence identity with 35 reference strains. The phylogenetic analysis demonstrated that the two PBoV strains CH/HNZM and PBoV-TY clustered into the PBoV2 group, and PBoV-TY was closely related to the PBoV strain GD18 strain (KJ755666) which may be derived from PBoV strains 0912/2012 (MH558677) and 57AT-HU (KF206160) through the recombination analysis. Compared with reference strain ZJD (HM053694)-China, a higher variation in the NS1 amino acids of PBoVs was found for CH/HNZM and PBoV-TY. These results had provided further information of the PBoV prevalence in central China, and they extend our understanding of the molecular epidemiology and evolution of PBoV.

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Concurrent infections of pseudorabies virus and porcine bocavirus in China detected by duplex nanoPCR

Cited by 14 publications

References 17 publications

Detection and genetic characteristic of porcine bocavirus in central China

Detection and genetic characteristic of porcine bocavirus in central China

Simultaneous Detection of Bovine Rotavirus, Bovine Parvovirus, and Bovine Viral Diarrhea Virus Using a Gold Nanoparticle-Assisted PCR Assay With a Dual-Priming Oligonucleotide System

Molecular detection, phylogenetic and evolutionary analysis of porcine bocavirus in central China

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