Development of droplet digital PCR for quantification of bovine leukemia virus proviral load using unpurified genomic DNA.
J Virol Methods
; 315: 114706, 2023 05.
Article
en En
| MEDLINE
| ID: mdl-36849053
ABSTRACT
Bovine leukemia virus (BLV) is the causative agent of a B-cell tumor called enzootic bovine leukosis. Preventing BLV spreading is required to reduce economic loss related to BLV infection of livestock. To quantify proviral load (PVL) more easily and rapidly, we developed a quantification system of PVL using droplet digital PCR (ddPCR). This method uses a multiplex TaqMan assay of the BLV provirus and housekeeping gene RPP30 for the quantification of BLV in BLV-infected cells. Furthermore, we combined ddPCR with DNA purification-free sample preparation (unpurified genomic DNA). The percentage of BLV-infected cells based on unpurified genomic DNA was highly correlated with that based on purified genomic DNA (correlation coefficient 0.906). Thus, this new technique is a suitable method to quantify PVL of BLV-infected cattle in a large sample number.
Palabras clave
Texto completo:
1
Banco de datos:
MEDLINE
Asunto principal:
Leucosis Bovina Enzoótica
/
Virus de la Leucemia Bovina
Tipo de estudio:
Diagnostic_studies
Límite:
Animals
Idioma:
En
Año:
2023
Tipo del documento:
Article