Multiply-primed rolling-circle amplification (MPRCA) of PCV2 genomes: Applications on detection, sequencing and virus isolation

Dezen, Diogenes; Rijsewijk, F.A.M.; Teixeira, Thaís Fumaco; Holz, Carine Lidiane; Cibulski, Samuel Paulo; Franco, Ana Cláudia; Dellagostin, Odir A.; Roehe, Paulo Michel

doi:10.1016/j.rvsc.2009.10.006

Cited by 17 publications

(12 citation statements)

References 25 publications

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“…Multiply primed rolling-circle amplification (RCA) was performed as previously described [ 10 , 13 ]. Briefly, 100 ng of total DNA (2 μL of 50 ng/μL solution) from papillomatous tissue was denatured at 95°C for 5 minutes and immediately cooled on ice.…”

Section: Methodsmentioning

confidence: 99%

“…The multiply primed rolling-circle amplification (RCA) strategy has been successfully used for the identification of the circular genomes of a number of viruses [ 8 ], e.g., anelloviruses [ 9 ], circoviruses [ 10 ] and papillomaviruses [ 11 ]. The method utilizes bacteriophage ϕ29 DNA polymerase for the selective amplification of circular DNA [ 8 ].…”

Section: Introductionmentioning

confidence: 99%

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Novel Bovine Papillomavirus Type Discovered by Rolling-Circle Amplification Coupled with Next-Generation Sequencing

et al. 2016

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Currently, fifteen bovine papillomavirus (BPV) types have been identified and classified into four genera: Deltapapillomavirus, Epsilonpapillomavirus, Dyoxipapillomavirus, and Xipapillomavirus. Here, the complete genome sequence of a new BPV type (BPV 04AC14) recovered from a papillomatous lesion is reported. The genome is 7,282 bp in length and exhibits the classic genetic organization and motifs of the members of Papillomaviridae. Maximum likelihood phylogenetic analyses revealed that BPV 04AC14 clusters with members of the Xipapillomavirus genus. The nucleotide sequence of the L1 capsid protein of the novel BPV is closely related to its counterpart, BPV3, with which it shares 79% similarity. These findings suggest that this virus is a new BPV type of the Xipapillomavirus genus.

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Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Novel Bovine Papillomavirus Type Discovered by Rolling-Circle Amplification Coupled with Next-Generation Sequencing

et al. 2016

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“…A rolling circle amplification (RCA) was applied to 100 ng of total DNA isolated from a papilloma lesion as previously described ( Dezen et al 2010 , Rijsewijk et al 2011 ). Neoplastic tissue was comprised by exophytic papillomatous, epithelium proliferation, and well-differentiated cells, marked acanthosis, koilocytes, increased amounts of granules in the granular layer, and keratohyalin granules.…”

mentioning

confidence: 99%

Complete genome sequence of Deltapapillomavirus 4 (bovine papillomavirus 2) from a bovine papillomavirus lesion in Amazon Region, Brazil

Daudt

Silva

Cibulski

et al. 2016

Mem. Inst. Oswaldo Cruz

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The complete genome sequence of bovine papillomavirus 2 (BPV2) from Brazilian Amazon Region was determined using multiple-primed rolling circle amplification followed by Illumina sequencing. The genome is 7,947 bp long, with 45.9% GC content. It encodes seven early (E1, E2,E4, E5, E6,E7, and E8) and two late (L1 and L2) genes. The complete genome of a BPV2 can help in future studies since this BPV type is highly reported worldwide although the lack of complete genome sequences available.

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“…In this study, the full-length of PCV2 from co-infected samples were amplified by PCR using a pair of primers, which could avoid artificial recombination events generating. In addition, nt sequencing of complete PCV2 genome obtained through rolling-circle amplification assay could overcome this problem [24,25]. …”

Section: Resultsmentioning

confidence: 99%

Co-existence of multiple strains of porcine circovirus type 2 in the same pig from China

Zhai

Chen

Wei

et al. 2011

Virol J

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Pigs are often co-infected by different viral strains from the same virus. Up to now, there are few reports about co-existence of different porcine circovirus type 2 (PCV2) strains in China. The aim of this study was to evaluate it in Chinese swine herds. 118 PCV2 positive DNAs isolated from diseased pigs identified by classic PCR were re-detected using a modified differential PCR assay. The results indicated that co-existence rates of PCV2 were 32.2% (38/118) in diseased pigs and 0% (0/41) in asymptomatic pigs. Four PCV2 complete genomes were cloned from two co-infected samples and their nucleotide (nt) identities were 95%-97.3%. The phylogenetic analysis showed that four PCV2 strains were divided into different genotypes, PCV2a, PCV2b, PCV2d and PCV2e, respectively. In addition, co-existence were not detected in 41 serum samples from healthy pigs but PCV2 single infection (31.7%, 13/41) existed. These data revealed that the co-existence of different strains of PCV2 might contribute to the development of more severe clinical symptoms for pigs. This is the first report confirming the co-existence of different PCV2 strains in Chinese swine herds. Meanwhile, this study could help us to understand new infection and prevalence forms of PCV2 clinically.

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Multiply-primed rolling-circle amplification (MPRCA) of PCV2 genomes: Applications on detection, sequencing and virus isolation

Cited by 17 publications

References 25 publications

Novel Bovine Papillomavirus Type Discovered by Rolling-Circle Amplification Coupled with Next-Generation Sequencing

Novel Bovine Papillomavirus Type Discovered by Rolling-Circle Amplification Coupled with Next-Generation Sequencing

Complete genome sequence of Deltapapillomavirus 4 (bovine papillomavirus 2) from a bovine papillomavirus lesion in Amazon Region, Brazil

Co-existence of multiple strains of porcine circovirus type 2 in the same pig from China

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