Comparative Evaluation of Three Commercial Quantitative Real-Time PCRs Used in Japan for Bovine Leukemia Virus
Bovine leukemia virus (BLV) is an oncogenic virus belonging to the genus <i>Deltaretrovirus</i> and is the causative agent of enzootic bovine leukosis. Proviral load (PVL) determined by real-time quantitative PCR (qPCR) is now widely used as an indicator of not only BLV infection, but al...
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| Format: | Article |
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MDPI AG
2022-05-01
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| Series: | Viruses |
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| Online Access: | https://www.mdpi.com/1999-4915/14/6/1182 |
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| author | Syuji Yoneyama Sota Kobayashi Towa Matsunaga Kaoru Tonosaki Dongze Leng Yusuke Sakai Shinji Yamada Atsushi Kimura Toshihiro Ichijo Hirokazu Hikono Kenji Murakami |
| author_facet | Syuji Yoneyama Sota Kobayashi Towa Matsunaga Kaoru Tonosaki Dongze Leng Yusuke Sakai Shinji Yamada Atsushi Kimura Toshihiro Ichijo Hirokazu Hikono Kenji Murakami |
| author_sort | Syuji Yoneyama |
| collection | DOAJ |
| description | Bovine leukemia virus (BLV) is an oncogenic virus belonging to the genus <i>Deltaretrovirus</i> and is the causative agent of enzootic bovine leukosis. Proviral load (PVL) determined by real-time quantitative PCR (qPCR) is now widely used as an indicator of not only BLV infection, but also BLV disease progression. To interpret PVLs determined by different qPCRs used in Japan, we compared a chimeric cycling probe-based qPCR, CY415, targeting the BLV tax region; a TaqMan probe-based qPCR, RC202, targeting the BLV pol region; and a TaqMan probe-based qPCR, CoCoMo, targeting the BLV long terminal repeat (LTR) region. Whole-blood samples collected from 317 naturally BLV-infected cattle (165 Holstein–Friesian and 152 Japanese Black) and tumor tissue samples collected from 32 cattle at a meat inspection center were used. The PVLs determined by each qPCR were strongly correlated. However, the PVL and the proportion of BLV-infected cells determined by RC202 or CoCoMo were significantly higher than those determined by CY415. Genetic analysis of three tumor tissue samples revealed that LTR region mutations or a deletion affected the PVL determined by CoCoMo. These results suggest that the TaqMan-based RC202 or CoCoMo qPCR is better than CY415 for BLV PVL analysis. However, qPCR target region mutations were not rare in tumors and could hamper PVL analysis by using qPCR. |
| first_indexed | 2024-03-09T22:15:10Z |
| format | Article |
| id | doaj.art-41766d57aef64200a158e5376ff1ee28 |
| institution | Directory Open Access Journal |
| issn | 1999-4915 |
| language | English |
| last_indexed | 2024-03-09T22:15:10Z |
| publishDate | 2022-05-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Viruses |
| spelling | doaj.art-41766d57aef64200a158e5376ff1ee282023-11-23T19:24:58ZengMDPI AGViruses1999-49152022-05-01146118210.3390/v14061182Comparative Evaluation of Three Commercial Quantitative Real-Time PCRs Used in Japan for Bovine Leukemia VirusSyuji Yoneyama0Sota Kobayashi1Towa Matsunaga2Kaoru Tonosaki3Dongze Leng4Yusuke Sakai5Shinji Yamada6Atsushi Kimura7Toshihiro Ichijo8Hirokazu Hikono9Kenji Murakami10Graduate School of Veterinary Sciences, Iwate University, Morioka 020-8550, JapanDivision of Zoonosis Research, National Institute of Animal Health, National Agriculture and Food Research Organization, Tsukuba 305-0856, JapanDepartment of Veterinary Sciences, Faculty of Agriculture, Iwate University, Morioka 020-8550, JapanDepartment of Plant Biosciences, Faculty of Agriculture, Iwate University, Morioka 020-8550, JapanGraduate School of Veterinary Sciences, Iwate University, Morioka 020-8550, JapanGraduate School of Veterinary Sciences, Iwate University, Morioka 020-8550, JapanGraduate School of Veterinary Sciences, Iwate University, Morioka 020-8550, JapanDepartment of Veterinary Sciences, Faculty of Agriculture, Iwate University, Morioka 020-8550, JapanDepartment of Veterinary Sciences, Faculty of Agriculture, Iwate University, Morioka 020-8550, JapanDepartment of Animal Sciences, Teikyo University of Science, Tokyo 120-0045, JapanGraduate School of Veterinary Sciences, Iwate University, Morioka 020-8550, JapanBovine leukemia virus (BLV) is an oncogenic virus belonging to the genus <i>Deltaretrovirus</i> and is the causative agent of enzootic bovine leukosis. Proviral load (PVL) determined by real-time quantitative PCR (qPCR) is now widely used as an indicator of not only BLV infection, but also BLV disease progression. To interpret PVLs determined by different qPCRs used in Japan, we compared a chimeric cycling probe-based qPCR, CY415, targeting the BLV tax region; a TaqMan probe-based qPCR, RC202, targeting the BLV pol region; and a TaqMan probe-based qPCR, CoCoMo, targeting the BLV long terminal repeat (LTR) region. Whole-blood samples collected from 317 naturally BLV-infected cattle (165 Holstein–Friesian and 152 Japanese Black) and tumor tissue samples collected from 32 cattle at a meat inspection center were used. The PVLs determined by each qPCR were strongly correlated. However, the PVL and the proportion of BLV-infected cells determined by RC202 or CoCoMo were significantly higher than those determined by CY415. Genetic analysis of three tumor tissue samples revealed that LTR region mutations or a deletion affected the PVL determined by CoCoMo. These results suggest that the TaqMan-based RC202 or CoCoMo qPCR is better than CY415 for BLV PVL analysis. However, qPCR target region mutations were not rare in tumors and could hamper PVL analysis by using qPCR.https://www.mdpi.com/1999-4915/14/6/1182bovine leukemia virusBLVcommercial kitprovirus loadqPCR |
| spellingShingle | Syuji Yoneyama Sota Kobayashi Towa Matsunaga Kaoru Tonosaki Dongze Leng Yusuke Sakai Shinji Yamada Atsushi Kimura Toshihiro Ichijo Hirokazu Hikono Kenji Murakami Comparative Evaluation of Three Commercial Quantitative Real-Time PCRs Used in Japan for Bovine Leukemia Virus Viruses bovine leukemia virus BLV commercial kit provirus load qPCR |
| title | Comparative Evaluation of Three Commercial Quantitative Real-Time PCRs Used in Japan for Bovine Leukemia Virus |
| title_full | Comparative Evaluation of Three Commercial Quantitative Real-Time PCRs Used in Japan for Bovine Leukemia Virus |
| title_fullStr | Comparative Evaluation of Three Commercial Quantitative Real-Time PCRs Used in Japan for Bovine Leukemia Virus |
| title_full_unstemmed | Comparative Evaluation of Three Commercial Quantitative Real-Time PCRs Used in Japan for Bovine Leukemia Virus |
| title_short | Comparative Evaluation of Three Commercial Quantitative Real-Time PCRs Used in Japan for Bovine Leukemia Virus |
| title_sort | comparative evaluation of three commercial quantitative real time pcrs used in japan for bovine leukemia virus |
| topic | bovine leukemia virus BLV commercial kit provirus load qPCR |
| url | https://www.mdpi.com/1999-4915/14/6/1182 |
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