PCR-based methods for detection of JSRV in experimentally and naturally infected sheep

• Autorzy: Kycko A. , Reichert M.
• Języki publikacji: EN

Abstrakty

EN

Application of real-time PCR using Taqman probe was tested for jaagsiekte sheep retrovirus (JSRV) detection. Sensitivity of real-time PCR and hemi-nested PCR methods was compared using plasmid DNA. The methods, along with RT PCR and real-time RT PCR, were tested for the possibility of JSRV genome (LTR region) detection in biological material from experimentally and naturally infected sheep. The experimental group of eight animals was used, including five lambs infected with JSRV by intratracheal inoculation at the age of 2 weeks. The samples collected from the animals ante-mortem included blood and respiratory tract fluid. Lung tissue, mediastinal lymph nodes, spleen, and liver were collected post-mortem. The field studies included blood samples collected from sheep from Polish flocks and lung samples obtained from slaughterhouse. In addition, DNA samples isolated from blood of sheep from the abroad located flocks with history of ovine pulmonary adenomatosis (OPA) were also included. Lung samples were examined histologically for the presence of pulmonary adenocarcinoma. The sensitivity of PCR, hemi-nested PCR, and real-time PCR using Taqman probe was evaluated as 10³, 10², and 10² viral copies, respectively. Both viral RNA and DNA were detected in the lung fluid taken from JSRV infected sheep showing clinical sings of OPA and in all neoplastic tissues. Proviral DNA was found in mediastinal lymph node of one experimental sheep. Five of the 66 DNA samples from the abroad located farms were positive for the presence of JSRV LTR. All blood and lung samples collected from Polish sheep were negative for the presence of JSRV LTR. The characteristic adenocarcinoma lesions were found in all lung sections of experimentally infected sheep. Implementation of the real-time PCR method is a good alternative to traditional PCR and hnPCR in JSRV detection and, apart from histopathological and immunohistochemical examinations, may be used as a confirmatory test in clinically suspected cases, or as a screening method in control or eradication scheme.

Słowa kluczowe

EN

polymerase chain reaction; detection; jaagsiekte sheep retrovirus; animal infection; naturally infected sheep; sheep; real-time polymerase chain reaction; adenocarcinoma

• Wydawca: -
• Czasopismo: Bulletin of the Veterinary Institute in Puławy
• Rocznik: 2010
• Tom: 54
• Numer: 4
• Opis fizyczny: p.445-450,fig.,ref.

Twórcy

autor

Kycko A.

• Department of Pathology, National Veterinary Research Institute, 24-100 Pulawy, Poland

autor

Reichert M.

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