Establishment of a simplified inverse polymerase chain reaction method for diagnosis of enzootic bovine leukosis

Enzootic bovine leukosis (EBL) is a malignant B-cell lymphoma of cattle caused by infection with bovine leukemia virus (BLV). It is defined by clonal and neoplastic expansion of BLV-infected B cells. Currently, multiple examinations are able to comprehensively diagnose this condition. Inverse polyme...

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Bibliographic Details
Published in:Archives of virology 2021-03, Vol.166 (3), p.841-851
Main Authors: Nishimori, Asami, Andoh, Kiyohiko, Matsuura, Yuichi, Kumagai, Asuka, Hatama, Shinichi
Format: Article
Language:English
Subjects:
Animals
B-cell lymphoma
B-Lymphocytes - pathology
B-Lymphocytes - virology
Biomedical and Life Sciences
Biomedicine
Bovine leukosis
Cattle
Cell Line, Tumor
Diagnosis
Enzootic Bovine Leukosis - diagnosis
Enzootic Bovine Leukosis - virology
Infectious Diseases
Integration
Leukemia
Leukemia Virus, Bovine - genetics
Leukosis
Lymphocytes B
Lymphoma, B-Cell - veterinary
Medical Microbiology
Original Article
Polymerase chain reaction
Polymerase Chain Reaction - methods
Proviruses - genetics
Reproducibility of Results
Sensitivity and Specificity
Virology
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Summary:Enzootic bovine leukosis (EBL) is a malignant B-cell lymphoma of cattle caused by infection with bovine leukemia virus (BLV). It is defined by clonal and neoplastic expansion of BLV-infected B cells. Currently, multiple examinations are able to comprehensively diagnose this condition. Inverse polymerase chain reaction (PCR) is a useful method to determine retrovirus integration sites. Here, we established a simplified inverse PCR method, involving the evaluation of clonality and similarity of BLV integration sites, to clinically diagnose EBL, and we also assessed its reliability. We found that the novel BLV inverse PCR could detect clonal expansion of infected cells even if they constituted only 5% of the total number of cells, while not amplifying any fragments from BLV-uninfected cells, thus confirming its sufficient sensitivity and specificity for use in EBL diagnosis. Furthermore, 50 clinical cases of bovine leukemia were analyzed using BLV inverse PCR and other PCR-based methods, wherein our method most efficiently determined virus-dependent bovine leukemia, including unidentified clinical cases observed in a previous report. Following further clinical investigations to enhance its reliability, the proposed BLV inverse PCR method has the potential to be applied to EBL diagnosis.
ISSN:0304-8608
1432-8798
DOI:10.1007/s00705-020-04945-4