Development of a direct blood-based PCR system to detect BLV provirus using CoCoMo primers.

Abstract:

:Bovine leukemia virus (BLV), the etiologic agent of enzootic bovine leucosis, has caused pandemic outbreaks worldwide. Because transcription of the BLV is quickly blocked after infection, detecting integrated provirus at host genome is an important method of identifying whether an animal is infected. The aim of the present study was to develop a novel direct blood-based PCR system to detect the BLV provirus with high specificity and at low cost. The assay was based on the BLV-CoCoMo degenerate primers, which amplify all known BLV strains. Cattle blood samples (n = 182) were collected from the same BLV-positive farm and subjected to BLV-CoCoMo-direct-PCR to detect the BLV provirus. The proviral load was then estimated. This novel PCR method showed 100 % specificity. The BLV-CoCoMo-direct-PCR can be used in a variety of laboratory situations because it does not require expensive equipment/reagents, DNA purification, or a second round of PCR. Therefore, the method is extremely cost-effective and the risk of a false-positive result due to DNA contamination is very low.

journal_name

Arch Virol

journal_title

Archives of virology

authors

Takeshima SN,Watanuki S,Ishizaki H,Matoba K,Aida Y

doi

10.1007/s00705-016-2806-y

subject

Has Abstract

pub_date

2016-06-01 00:00:00

pages

1539-46

issue

6

eissn

0304-8608

issn

1432-8798

pii

10.1007/s00705-016-2806-y

journal_volume

161

pub_type

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