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Dr. Hala Gamal Ali Ali El Daous :: Publications: |
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| Title: | Development of droplet digital PCR for quantification of bovine leukemia virus proviral load using unpurified genomic DNA |
| Authors: | Xinyue Wu a, Kousuke Notsu a, Yuichi Matsuura b, Shuya Mitoma c, Hala El Daous d, Junzo Noriminee,f, Satoshi Sekiguchie,f,* |
| Year: | 2023 |
| Keywords: | Bovine leukemia virus Diagnostic test Droplet digital PCR Enzootic bovine leukosis Quantification Unpurified genomic DNA |
| Journal: | Journal of Virological Methods |
| Volume: | Not Available |
| Issue: | Not Available |
| Pages: | Not Available |
| Publisher: | Elsevier |
| Local/International: | International |
| Paper Link: | Not Available |
| Full paper | Hala Gamal Ali Ali El Daous_1-s2.0-S0166093423000319-main.pdf |
| Supplementary materials | Not Available |
| 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 quan- tification 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. |
