Nested-multiplex PCR detection of Orthopoxvirus and Parapoxvirus directly from exanthematic clinical samples

Abrahão, Jônatas Santos; Lima, Luciano Soares de; Assis, Felipe Lopes de; Alves, Pedro Augusto; Silva‐Fernandes, André Tavares da; Cota, Marcela Menezes Gomes; Ferreira, Vinícius V. M.; Campos, Rafael K.; Mazur, Carlos; Lobato, Zélia Inês Portela; Trindade, Giliane S.; Kroon, Erna Geessien

doi:10.1186/1743-422x-6-140

Cited by 40 publications

(41 citation statements)

References 19 publications

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“…Nucleic acids were isolated with a commercial kit (High Pure Viral Nucleic Acid Extraction Kit, Roche, Germany). A nested polymerase chain reaction (PCR) protocol was used for detection of parapoxvirus DNA in samples [8]. The PCR products were purified by using EZ-10 Spin Column Gel Extraction kit (Bio Basic, Ontario, Canada) and sequenced bi-directionally on ABI-PRISM 310 Genetic Analyzer, using BigDye chemistry (Applied Biosystems, CA, USA).…”

Section: Outbreak Investigation and Resultsmentioning

confidence: 99%

Nosocomial outbreak of disseminated orf infection in a burn unit, Gaziantep, Turkey, October to December 2012

et al. 2013

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We report the first outbreak of nosocomial orf infection in a hospital burn unit in Gaziantep, Turkey. The outbreak lasted from October to December 2012 and involved a total of thirteen cases. It demonstrates the risk of introduction of orf virus to a burn unit, and the potential for extensive transmission among patients with compromised skin integrity. The importance of hygiene measures and infection control are highlighted and possible transmission routes of the virus discussed.

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Section: Outbreak Investigation and Resultsmentioning

confidence: 99%

Nosocomial outbreak of disseminated orf infection in a burn unit, Gaziantep, Turkey, October to December 2012

et al. 2013

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“…13 Total DNA extracted from baby hamster kidney (BHK)-21 cells infected with the ORFV strain IA-82 9 was used as a positive control. 3 As the set of primers was expected to amplify all PPVs, 13 the amplicons were sequenced to identify the specific PPV. The PCR products were purified using a commercial kit a according to the manufacturer's instructions and sequenced.…”

mentioning

confidence: 99%

An outbreak of pseudocowpox in fattening calves in southern Brazil

et al. 2012

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Pseudocowpox virus is a parapoxvirus frequently associated with papulovesicular and scabby lesions on the teats and udders of milking cows and is often transmitted to human beings. An unusual outbreak of skin disease in fattening calves in southern Brazil is described. Fourteen of 17 male cattle (82%), aged 6–48 months, feeding on grass pastures were affected. Animals developed papules, vesicles, and scabby proliferative lesions on the muzzle in a clinical course of approximately 10–15 days. The scabby lesions often presented with exudation and bleeding. Histological examination of mucocutaneous tissue in detached scabs revealed acanthosis with thickening of the corneal layer and premature keratinization (parakeratotic hyperkeratosis). The dermis had multifocal lymphoplasmacytic infiltrates. Electron microscopic examination of scab specimens revealed typical parapoxvirus particles: oval shaped (260 nm × 160 nm), enveloped, and covered with a helical layer. Polymerase chain reaction using a set of pan-parapoxvirus primers for the B2L gene amplified a 590-bp product out of DNA extracted from scabs. Nucleotide sequencing of the amplicons revealed a nucleotide homology of 97% with Pseudocowpox virus and lower homology with other parapoxviruses: Bovine papular stomatitis virus (84%) and Orf virus (94%). A phylogenetic tree based on the B2L sequence was constructed, showing that the virus clustered with Pseudocowpox virus isolates.

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“…The scabs were macerated using a homogenizer (Politron, Littau, Switzerland) in PBS, which contained 200 U of penicillin, 4 lg of amphotericin B, and 100 lg of gentamicin per mL (0.1 g scab/0.9 mL PBS), and clarified by centrifugation at 20009g for 3 min [13]. The supernatants from scabs and diluted sera were used for diagnostic purposes, viral isolation, and molecular assays.…”

Section: Molecular Diagnosismentioning

confidence: 99%

Horizontal study of vaccinia virus infections in an endemic area: epidemiologic, phylogenetic and economic aspects

Assis

Franco-Luiz

Paim³

et al. 2015

Arch Virol

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Vaccinia virus (VACV), the etiological agent of bovine vaccinia (BV), is widespread in Brazil and present in most of the milk-producing regions. We conducted a horizontal study of BV in Bahia, a state of Brazil in which the production of milk is increasing. During 2011, human and bovine clinical samples were collected during outbreaks for BV diagnosis, virus isolation and molecular analysis. We collected data for epidemiological inferences. Vaccinia virus was detected in 87.7% of the analyzed outbreaks, highlighting the effective circulation of VACV in Bahia. The molecular data showed the spreading of group 1 Brazilian VACV to Bahia. We observed a seasonal profile of BV, with its peak in the drier and cooler season. Manual milking was observed in 96 % of the visited properties, showing its importance to viral spread in herds. Under-notification of BV, ineffective animal trade surveillance, and bad milking practices have contributed to the spread of VACV in Brazil.

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Nested-multiplex PCR detection of Orthopoxvirus and Parapoxvirus directly from exanthematic clinical samples

Cited by 40 publications

References 19 publications

Nosocomial outbreak of disseminated orf infection in a burn unit, Gaziantep, Turkey, October to December 2012

Nosocomial outbreak of disseminated orf infection in a burn unit, Gaziantep, Turkey, October to December 2012

An outbreak of pseudocowpox in fattening calves in southern Brazil

Horizontal study of vaccinia virus infections in an endemic area: epidemiologic, phylogenetic and economic aspects

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