A nested PCR approach for unambiguous typing of pestiviruses infecting cattle

Decaro, Nicola; Sciarretta, Rossana; Lucente, Maria Stella; Mari, Viviana; Amorisco, Francesca; Colaianni, Maria Loredana; Cordioli, Paolo; Parisi, Antonio; Lelli, Rossella; Buonavoglia, Canio

doi:10.1016/j.mcp.2011.11.003

Cited by 29 publications

(43 citation statements)

References 36 publications

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“…Detection of pestiviruses in biological fluids and tissue samples from the dead calf was carried out with a real-time reverse transcription (RT)-PCR assay specific for Hobi-like pestivirus (20) and a nested PCR protocol for simultaneous detection and characterization of all of the pestiviruses that infect cattle (21).…”

Section: Methodsmentioning

confidence: 99%

Mucosal Disease-Like Syndrome in a Calf Persistently Infected by Hobi-Like Pestivirus

et al. 2014

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A calf persistently infected with Hobi-like pestivirus displayed severe clinical signs and subsequently died. Gross lesions and histopathological changes were suggestive of hemorrhagic and necrotic inflammation involving several tissues. A Hobi-like pestivirus pair was isolated from the dead calf, i.e., cytopathogenic (CP) and noncytopathogenic (NCP) strains strictly related to each other and to Italian prototype isolates at the genetic level. Two biotype-specific real-time reverse transcription-PCR assays determined the time of the emergence of the CP virus as 1 month before the calf's death. This highest RNA titers were reached in lymphoid and nervous system tissues, whereas only traces of CP viral RNA were found in blood. In contrast, great NCP virus loads were present in all tissues and biological fluids. The present report provides new insights into the pathogenesis and molecular mechanisms of this emerging group of pestiviruses.

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Section: Methodsmentioning

confidence: 99%

Mucosal Disease-Like Syndrome in a Calf Persistently Infected by Hobi-Like Pestivirus

et al. 2014

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“…The farmer was convinced to carry out an eradication programme aimed to detect and slaughter PI animals. For this purpose, all the 752 animals of the herd were bled by the farm personnel and EDTA blood samples were submitted to nested‐PCR (Decaro et al., ) and real‐time RT‐PCR assays (Losurdo et al., ; Mari et al., ) for detection and characterization of ruminant pestiviruses. A total of 9 PI animals were detected that were immediately slaughtered.…”

Section: Case Reportmentioning

confidence: 99%

HoBi-Like Pestivirus and Its Impact on Cattle Productivity

Decaro

Lucente

Losurdo

et al. 2016

Transbound Emerg Dis

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The clinical features and economic impact of the infection caused by an emerging group of pestiviruses, namely HoBi-like pestivirus, in a cattle herd of southern Italy are reported. In 2011, the virus was first associated with respiratory disease, causing an abortion storm after 1 year and apparently disappearing for the following 3 years after persistently infected calves were slaughtered. However, in 2014, reproductive failures and acute gastroenteritis were observed in the same herd, leading to a marked decrease of productivity. A HoBi-like strain closely related to that responsible for previous outbreaks was detected in several animals. Application of an intensive eradication programme, based on the detection and slaughtering of HoBi-like pestivirus persistently infected animals, resulted in a marked improvement of the productive performances.

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“…(1), and bovid herpesvirus 1 (26) and by RT-PCR for bluetongue virus (3,8). Conversely, the pestivirus genome was detected with two different RT-PCR protocols (7,24), and the virus was characterized as Hobi-like by a species-specific nested PCR (7) (Fig. 1A).…”

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Hobi-Like Pestivirus in Aborted Bovine Fetuses

Decaro¹,

Lucente²,

Mari³

et al. 2012

J Clin Microbiol

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An outbreak of abortion affecting multiparous cows was associated with Hobi-like pestivirus infection. Viral RNA and antigens were detected in the tissues of two aborted fetuses. Molecular assays for other common abortogenic agents tested negative. At the genetic level, the Hobi-like pestivirus displayed the closest relatedness to Italian, Australian, and South American viruses, whereas it diverged from the prototype Thai isolate. These findings may have important implications for the pestivirus control/ eradication programs in cattle herds. Bovine viral diarrhea viruses (BVDVs) are members of the genus Pestivirus (family Flaviviridae), responsible for a number of clinical signs, including subclinical infections, immunosuppression, acute diarrhea, respiratory disease, reproductive failures, and mucosal disease in persistently infected calves. Reproductive disorders caused by BVDVs vary according to the fetal age and include embryo death, abortion, mummification, congenital abnormalities, or stillbirths (2). Thus far, two different BVDV species have been recognized, BVDV-1 and BVDV-2, that cocirculate in cattle herds worldwide (20). In 2004, an atypical pestivirus, strain D32/00_Hobi, was isolated from a contaminated batch of fetal calf serum (FCS) (19). The virus was distantly related to BVDV-1/BVDV-2, and it was proposed as a prototype of a new pestivirus species (14, 15). Hobi-like sequences have been repeatedly detected in commercial FCS batches (17,22,23), whereas there are few reports on natural infections (4, 5, 21, 23) and clinical outbreaks (5). The virus has been recently detected in aborted bovine fetuses in Brazil, thus suggesting direct clinical implications (4).Here, we report the isolation and genetic characterization of a Hobi-like strain detected from aborted fetuses in southern Italy. The abortion outbreak occurred in June 2011 in a cattle herd where a Hobi-like pestivirus-associated respiratory disease had been recently described (5). Abortion was observed in eight multiparous cows in a group of 270 lactating Holstein cows and occurred between the fourth and sixth months of pregnancy. The animals neither showed prodromal signs nor presented postabortion complications. Two aborted fetuses (280/11-A, 280/11-B) were sent to our laboratory, and tissue samples were collected from lungs, spleens, livers, kidneys, and placentas for diagnostic investigations. Nucleic acids were purified using the DNeasy tissue kit (Qiagen) and QIAamp RNeasy minikit (Qiagen). Reverse transcription (RT)-PCR and PCR assays were performed using SuperScript one-step RT-PCR for long templates (Life Technologies) and LA PCR kit version 2.1 (TaKaRa Bio Inc.), respectively. Positive and negative controls were processed in parallel to the screened samples. The samples tested negative by PCR for Chlamydophila spp. 3, 8). Conversely, the pestivirus genome was detected with two different 24), and the virus was characterized as Hobi-like by a species-specific nested PCR (7) (Fig. 1A). Viral titers, quantified by a TaqMan-based real...

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A nested PCR approach for unambiguous typing of pestiviruses infecting cattle

Cited by 29 publications

References 36 publications

Mucosal Disease-Like Syndrome in a Calf Persistently Infected by Hobi-Like Pestivirus

Mucosal Disease-Like Syndrome in a Calf Persistently Infected by Hobi-Like Pestivirus

HoBi-Like Pestivirus and Its Impact on Cattle Productivity

Hobi-Like Pestivirus in Aborted Bovine Fetuses

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