A novel universal probe library quantitative reverse transcription polymerase chain reaction method to profile immunological gene expression in blood of captive beluga whales ( Delphinapterusleucas)
Cytokines are fundamental for a functioning immune system, and thus, potentially serve as important indicators of animal health. Quantitation of mRNA using quantitative reverse transcription polymerase chain reaction (qRT-PCR) is an established immunological technique. It is particularly suitable fo...
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crpeerj:10.7287/peerj.preprints.2735 2024-06-02T08:04:16+00:00 A novel universal probe library quantitative reverse transcription polymerase chain reaction method to profile immunological gene expression in blood of captive beluga whales ( Delphinapterusleucas) Tsai, Ming-An Chen, I-Hua Wang, Jiann-Hsiung Chou, Shih-Jen Li, Tsung-Hsien Leu, Ming-Yih Ho, Hsiao-Kuan Yang, Wei Cheng 2017 http://dx.doi.org/10.7287/peerj.preprints.2735 https://peerj.com/preprints/2735.pdf https://peerj.com/preprints/2735.xml https://peerj.com/preprints/2735.html unknown PeerJ http://creativecommons.org/licenses/by/4.0/ posted-content 2017 crpeerj https://doi.org/10.7287/peerj.preprints.2735 2024-05-07T14:14:40Z Cytokines are fundamental for a functioning immune system, and thus, potentially serve as important indicators of animal health. Quantitation of mRNA using quantitative reverse transcription polymerase chain reaction (qRT-PCR) is an established immunological technique. It is particularly suitable for detecting the expression of proteins against which monoclonal antibodies are not available. In this study, we developed a novel probe-based quantitative gene expression assay for immunological assessment of captive beluga whales ( Delphinapterusleucas ) that is one of the most common cetacean species on display in aquariums worldwide. Six immunologically relevant genes (IL-2Rα, -4, -10, -12, TNFα, and IFNγ) were selected for analysis, and two validated housekeeping genes (PGK1 and RPL4) with stable expression were used as reference genes. Sixteen blood samples were obtained from four animals with different health conditions and stored in RNAlater solution. These samples were used for RNA extraction followed by qRT-PCR analysis. Analysis of gene transcripts was performed by relative quantitation using the comparative Cq method with the integration of amplification efficiency and two reference genes. The expression levels of each gene in the samples from clinically healthy animals were normally distributed. Transcript outliers for IL-2Rα, IL-4, IL-12, TNFα, and IFNγ were noticed in four samples collected from two clinically unhealthy animals. This assay has the potential to identify immune system deviation from normal state, which is caused by health problems. Furthermore, knowing the immune status of captive cetaceans could help both trainers and veterinarians in implementing preventive approaches prior to disease onset. Other/Unknown Material Beluga Beluga* PeerJ Publishing |
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Cytokines are fundamental for a functioning immune system, and thus, potentially serve as important indicators of animal health. Quantitation of mRNA using quantitative reverse transcription polymerase chain reaction (qRT-PCR) is an established immunological technique. It is particularly suitable for detecting the expression of proteins against which monoclonal antibodies are not available. In this study, we developed a novel probe-based quantitative gene expression assay for immunological assessment of captive beluga whales ( Delphinapterusleucas ) that is one of the most common cetacean species on display in aquariums worldwide. Six immunologically relevant genes (IL-2Rα, -4, -10, -12, TNFα, and IFNγ) were selected for analysis, and two validated housekeeping genes (PGK1 and RPL4) with stable expression were used as reference genes. Sixteen blood samples were obtained from four animals with different health conditions and stored in RNAlater solution. These samples were used for RNA extraction followed by qRT-PCR analysis. Analysis of gene transcripts was performed by relative quantitation using the comparative Cq method with the integration of amplification efficiency and two reference genes. The expression levels of each gene in the samples from clinically healthy animals were normally distributed. Transcript outliers for IL-2Rα, IL-4, IL-12, TNFα, and IFNγ were noticed in four samples collected from two clinically unhealthy animals. This assay has the potential to identify immune system deviation from normal state, which is caused by health problems. Furthermore, knowing the immune status of captive cetaceans could help both trainers and veterinarians in implementing preventive approaches prior to disease onset. |
format |
Other/Unknown Material |
author |
Tsai, Ming-An Chen, I-Hua Wang, Jiann-Hsiung Chou, Shih-Jen Li, Tsung-Hsien Leu, Ming-Yih Ho, Hsiao-Kuan Yang, Wei Cheng |
spellingShingle |
Tsai, Ming-An Chen, I-Hua Wang, Jiann-Hsiung Chou, Shih-Jen Li, Tsung-Hsien Leu, Ming-Yih Ho, Hsiao-Kuan Yang, Wei Cheng A novel universal probe library quantitative reverse transcription polymerase chain reaction method to profile immunological gene expression in blood of captive beluga whales ( Delphinapterusleucas) |
author_facet |
Tsai, Ming-An Chen, I-Hua Wang, Jiann-Hsiung Chou, Shih-Jen Li, Tsung-Hsien Leu, Ming-Yih Ho, Hsiao-Kuan Yang, Wei Cheng |
author_sort |
Tsai, Ming-An |
title |
A novel universal probe library quantitative reverse transcription polymerase chain reaction method to profile immunological gene expression in blood of captive beluga whales ( Delphinapterusleucas) |
title_short |
A novel universal probe library quantitative reverse transcription polymerase chain reaction method to profile immunological gene expression in blood of captive beluga whales ( Delphinapterusleucas) |
title_full |
A novel universal probe library quantitative reverse transcription polymerase chain reaction method to profile immunological gene expression in blood of captive beluga whales ( Delphinapterusleucas) |
title_fullStr |
A novel universal probe library quantitative reverse transcription polymerase chain reaction method to profile immunological gene expression in blood of captive beluga whales ( Delphinapterusleucas) |
title_full_unstemmed |
A novel universal probe library quantitative reverse transcription polymerase chain reaction method to profile immunological gene expression in blood of captive beluga whales ( Delphinapterusleucas) |
title_sort |
novel universal probe library quantitative reverse transcription polymerase chain reaction method to profile immunological gene expression in blood of captive beluga whales ( delphinapterusleucas) |
publisher |
PeerJ |
publishDate |
2017 |
url |
http://dx.doi.org/10.7287/peerj.preprints.2735 https://peerj.com/preprints/2735.pdf https://peerj.com/preprints/2735.xml https://peerj.com/preprints/2735.html |
genre |
Beluga Beluga* |
genre_facet |
Beluga Beluga* |
op_rights |
http://creativecommons.org/licenses/by/4.0/ |
op_doi |
https://doi.org/10.7287/peerj.preprints.2735 |
_version_ |
1800748900169023488 |