Cargando…
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 Deltaretrovirus 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 prog...
Autores principales: | , , , , , , , , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MDPI
2022
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9230052/ https://www.ncbi.nlm.nih.gov/pubmed/35746654 http://dx.doi.org/10.3390/v14061182 |
_version_ | 1784734961823121408 |
---|---|
author | Yoneyama, Syuji Kobayashi, Sota Matsunaga, Towa Tonosaki, Kaoru Leng, Dongze Sakai, Yusuke Yamada, Shinji Kimura, Atsushi Ichijo, Toshihiro Hikono, Hirokazu Murakami, Kenji |
author_facet | Yoneyama, Syuji Kobayashi, Sota Matsunaga, Towa Tonosaki, Kaoru Leng, Dongze Sakai, Yusuke Yamada, Shinji Kimura, Atsushi Ichijo, Toshihiro Hikono, Hirokazu Murakami, Kenji |
author_sort | Yoneyama, Syuji |
collection | PubMed |
description | Bovine leukemia virus (BLV) is an oncogenic virus belonging to the genus Deltaretrovirus 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. |
format | Online Article Text |
id | pubmed-9230052 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
spelling | pubmed-92300522022-06-25 Comparative Evaluation of Three Commercial Quantitative Real-Time PCRs Used in Japan for Bovine Leukemia Virus Yoneyama, Syuji Kobayashi, Sota Matsunaga, Towa Tonosaki, Kaoru Leng, Dongze Sakai, Yusuke Yamada, Shinji Kimura, Atsushi Ichijo, Toshihiro Hikono, Hirokazu Murakami, Kenji Viruses Article Bovine leukemia virus (BLV) is an oncogenic virus belonging to the genus Deltaretrovirus 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. MDPI 2022-05-28 /pmc/articles/PMC9230052/ /pubmed/35746654 http://dx.doi.org/10.3390/v14061182 Text en © 2022 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/). |
spellingShingle | Article Yoneyama, Syuji Kobayashi, Sota Matsunaga, Towa Tonosaki, Kaoru Leng, Dongze Sakai, Yusuke Yamada, Shinji Kimura, Atsushi Ichijo, Toshihiro Hikono, Hirokazu Murakami, Kenji Comparative Evaluation of Three Commercial Quantitative Real-Time PCRs Used in Japan for Bovine Leukemia Virus |
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 | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9230052/ https://www.ncbi.nlm.nih.gov/pubmed/35746654 http://dx.doi.org/10.3390/v14061182 |
work_keys_str_mv | AT yoneyamasyuji comparativeevaluationofthreecommercialquantitativerealtimepcrsusedinjapanforbovineleukemiavirus AT kobayashisota comparativeevaluationofthreecommercialquantitativerealtimepcrsusedinjapanforbovineleukemiavirus AT matsunagatowa comparativeevaluationofthreecommercialquantitativerealtimepcrsusedinjapanforbovineleukemiavirus AT tonosakikaoru comparativeevaluationofthreecommercialquantitativerealtimepcrsusedinjapanforbovineleukemiavirus AT lengdongze comparativeevaluationofthreecommercialquantitativerealtimepcrsusedinjapanforbovineleukemiavirus AT sakaiyusuke comparativeevaluationofthreecommercialquantitativerealtimepcrsusedinjapanforbovineleukemiavirus AT yamadashinji comparativeevaluationofthreecommercialquantitativerealtimepcrsusedinjapanforbovineleukemiavirus AT kimuraatsushi comparativeevaluationofthreecommercialquantitativerealtimepcrsusedinjapanforbovineleukemiavirus AT ichijotoshihiro comparativeevaluationofthreecommercialquantitativerealtimepcrsusedinjapanforbovineleukemiavirus AT hikonohirokazu comparativeevaluationofthreecommercialquantitativerealtimepcrsusedinjapanforbovineleukemiavirus AT murakamikenji comparativeevaluationofthreecommercialquantitativerealtimepcrsusedinjapanforbovineleukemiavirus |