Establishment of a novel diagnostic test for Bovine leukaemia virus infection using direct filter PCR

Abstract

Enzootic bovine leucosis (EBL) is a neoplastic disease caused by Bovine leukaemia virus (BLV) (Bartlett et al., 2014). BLV infection occurs in three possible stages: the aleukaemic stage, persistent lymphocytosis (PL) stage and EBL stage (Dequiedt et al., 1999). It is estimated that approximately 30% of BLV-infected cattle are in the PL stage, and fewer than 5% of those develop fatal B-cell lymphosarcoma (Rodríguez et al., 2011). This disease has an economic impact because it causes direct economic losses by reducing milk production and fertility, and through cattle condemnation and death (Kale et al., 2007). A great amount of efforts was directed to BLV elimination during the period 1993-2010 (EFSA Panel on Animal Health and Welfare (AHAW) et al., 2017) in the European Union (EU). As a result, most of its member states have officially declared EBL-free. In Japan, on the other hand, the nationwide survey of BLV infection from 2009 to 2011 showed that the prevalence of BLV infection in animal level was 40.9%

Dried Blood Spots technology for veterinary applications and biological investigations: technical aspects, retrospective analysis, ongoing status and future perspectives

“…Summarising literature data of the reported effectiveness of FTA cards for storage and shipment of viral RNA, Cardona-Ospina et al ( 2019 ) concluded that the cards could be a suitable option for these purposes in areas where proper conditions for RNA preservation are difficult to achieve. Different DNA viruses usually demonstrated good storage stability in dried samples, for example, avian Marek’s disease virus (Cortes et al 2009 ), duck hepatitis B virus (Wang et al 2002 ) and fowl adenovirus I (Moscoso et al 2007 ), bovine herpesvirus-1 (Sarangi et al 2018 ) and bovine leucosis virus (Saushkin et al 2019 ; El Daous et al 2020 ), African swine fever virus (Michaud et al 2007 ; Uttenthal et al 2013 ; Randriamparany et al 2014 ). Common filter papers are cheap and could be as good as more expensive commercial FTA cards for virus storage, like it was shown for African swine fever virus (Uttenthal et al 2013 ; Randriamparany et al 2014 ), infectious bursal disease virus (Moscoso et al 2006 ; Maw et al 2006 ) and Marek’s disease virus (Cortes et al 2009 ; Wannaratana et al 2021 ).…”

Bovine Leukaemia Virus: Current Epidemiological Circumstance and Future Prospective

“…This method is of lower sensitivity than nested PCR but indicates higher specificity and reproducibility, and is cost-effective as it is neither labour-intensive nor time-consuming [11]. Further, a direct filter PCR test has been established recently as a novel, fast, smooth, reliable, and practical diagnostic test that directly detects the BLV proviral DNA in clinical blood samples without DNA extraction while offering simple collection, transportation and storage procedures for clinical blood specimens [184].…”

Improving of the Nested PCR for Detection of Bovine Leukemia Virus

“…In the last years topicality of the BLV is increasing in view of new information related to human breast cancer [21]. It leads to new research in this area and the development of new methods of BLV identification in cattle to eradicate infected animals [17,18,[22][23][24][25][26].…”