Development and Visualization Improvement for the Rapid Detection of Decapod Iridescent Virus 1 (DIV1) in <i<Penaeus vannamei</i< Based on an Isothermal Recombinase Polymerase Amplification Assay
Viral diseases have seriously restricted the healthy development of aquaculture, and decapod iridescent virus 1 (DIV1) has led to heavy losses in the global shrimp aquaculture industry. Due to the lack of effective treatment, early detection and regular monitoring are the most effective ways to avoi...
Ausführliche Beschreibung
Autor*in: |
Yajin Xu [verfasserIn] Yan Wang [verfasserIn] Jingjie Hu [verfasserIn] Zhenmin Bao [verfasserIn] Mengqiang Wang [verfasserIn] |
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Format: |
E-Artikel |
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Sprache: |
Englisch |
Erschienen: |
2022 |
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Schlagwörter: |
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Übergeordnetes Werk: |
In: Viruses - MDPI AG, 2009, 14(2022), 12, p 2752 |
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Übergeordnetes Werk: |
volume:14 ; year:2022 ; number:12, p 2752 |
Links: |
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DOI / URN: |
10.3390/v14122752 |
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Katalog-ID: |
DOAJ082954917 |
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10.3390/v14122752 doi (DE-627)DOAJ082954917 (DE-599)DOAJfc8fe03f1b43447d849f801abe2780a3 DE-627 ger DE-627 rakwb eng QR1-502 Yajin Xu verfasserin aut Development and Visualization Improvement for the Rapid Detection of Decapod Iridescent Virus 1 (DIV1) in <i<Penaeus vannamei</i< Based on an Isothermal Recombinase Polymerase Amplification Assay 2022 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier Viral diseases have seriously restricted the healthy development of aquaculture, and decapod iridescent virus 1 (DIV1) has led to heavy losses in the global shrimp aquaculture industry. Due to the lack of effective treatment, early detection and regular monitoring are the most effective ways to avoid infection with DIV1. In this study, a novel real-time quantitative recombinase polymerase amplification (qRPA) assay and its instrument-free visualization improvement were described for the rapid detection of DIV1. Optimum primer pairs, suitable reaction temperatures, and probe concentrations of a DIV1-qRPA assay were screened to determine optimal reaction conditions. Then, its ability to detect DIV1 was evaluated and compared with real-time quantitative polymerase chain reactions (qPCRs). The sensitivity tests demonstrated that the limit of detection (LOD) of the DIV1-qRPA assay was 1.0 copies μL<sup<−1</sup<. Additionally, the presentation of the detection results was improved with SYBR Green I, and the LOD of the DIV1-RPA-SYBR Green I assay was 1.0 × 10<sup<3</sup< copies μL<sup<−1</sup<. Both the DIV1-qRPA and DIV1-RPA-SYBR Green I assays could be performed at 42 °C within 20 min and without cross-reactivity with the following: white spot syndrome virus (WSSV), <i<Vibrio parahaemolyticus</i< associated with acute hepatopancreatic necrosis disease (<i<V<sub<p</sub<</i<<sub<AHPND</sub<), <i<Enterocytozoon hepatopenaei</i< (EHP), and infectious hypodermal and hematopoietic necrosis virus (IHHNV). In conclusion, this approach yields rapid, straightforward, and simple DIV1 diagnoses, making it potentially valuable as a reliable tool for the detection and prevention of DIV1, especially where there is a paucity of laboratory equipment. decapod iridescent virus 1 recombinase polymerase amplification <i<Penaeus vannamei</i< white leg shrimp Microbiology Yan Wang verfasserin aut Jingjie Hu verfasserin aut Zhenmin Bao verfasserin aut Mengqiang Wang verfasserin aut In Viruses MDPI AG, 2009 14(2022), 12, p 2752 (DE-627)609775871 (DE-600)2516098-9 19994915 nnns volume:14 year:2022 number:12, p 2752 https://doi.org/10.3390/v14122752 kostenfrei https://doaj.org/article/fc8fe03f1b43447d849f801abe2780a3 kostenfrei https://www.mdpi.com/1999-4915/14/12/2752 kostenfrei https://doaj.org/toc/1999-4915 Journal toc kostenfrei GBV_USEFLAG_A SYSFLAG_A GBV_DOAJ GBV_ILN_11 GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_39 GBV_ILN_40 GBV_ILN_60 GBV_ILN_62 GBV_ILN_63 GBV_ILN_65 GBV_ILN_69 GBV_ILN_70 GBV_ILN_73 GBV_ILN_74 GBV_ILN_95 GBV_ILN_105 GBV_ILN_110 GBV_ILN_151 GBV_ILN_161 GBV_ILN_170 GBV_ILN_206 GBV_ILN_213 GBV_ILN_224 GBV_ILN_230 GBV_ILN_285 GBV_ILN_293 GBV_ILN_370 GBV_ILN_602 GBV_ILN_2005 GBV_ILN_2009 GBV_ILN_2011 GBV_ILN_2014 GBV_ILN_2055 GBV_ILN_2111 GBV_ILN_4012 GBV_ILN_4037 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4249 GBV_ILN_4305 GBV_ILN_4306 GBV_ILN_4307 GBV_ILN_4313 GBV_ILN_4322 GBV_ILN_4323 GBV_ILN_4324 GBV_ILN_4325 GBV_ILN_4338 GBV_ILN_4367 GBV_ILN_4700 AR 14 2022 12, p 2752 |
spelling |
10.3390/v14122752 doi (DE-627)DOAJ082954917 (DE-599)DOAJfc8fe03f1b43447d849f801abe2780a3 DE-627 ger DE-627 rakwb eng QR1-502 Yajin Xu verfasserin aut Development and Visualization Improvement for the Rapid Detection of Decapod Iridescent Virus 1 (DIV1) in <i<Penaeus vannamei</i< Based on an Isothermal Recombinase Polymerase Amplification Assay 2022 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier Viral diseases have seriously restricted the healthy development of aquaculture, and decapod iridescent virus 1 (DIV1) has led to heavy losses in the global shrimp aquaculture industry. Due to the lack of effective treatment, early detection and regular monitoring are the most effective ways to avoid infection with DIV1. In this study, a novel real-time quantitative recombinase polymerase amplification (qRPA) assay and its instrument-free visualization improvement were described for the rapid detection of DIV1. Optimum primer pairs, suitable reaction temperatures, and probe concentrations of a DIV1-qRPA assay were screened to determine optimal reaction conditions. Then, its ability to detect DIV1 was evaluated and compared with real-time quantitative polymerase chain reactions (qPCRs). The sensitivity tests demonstrated that the limit of detection (LOD) of the DIV1-qRPA assay was 1.0 copies μL<sup<−1</sup<. Additionally, the presentation of the detection results was improved with SYBR Green I, and the LOD of the DIV1-RPA-SYBR Green I assay was 1.0 × 10<sup<3</sup< copies μL<sup<−1</sup<. Both the DIV1-qRPA and DIV1-RPA-SYBR Green I assays could be performed at 42 °C within 20 min and without cross-reactivity with the following: white spot syndrome virus (WSSV), <i<Vibrio parahaemolyticus</i< associated with acute hepatopancreatic necrosis disease (<i<V<sub<p</sub<</i<<sub<AHPND</sub<), <i<Enterocytozoon hepatopenaei</i< (EHP), and infectious hypodermal and hematopoietic necrosis virus (IHHNV). In conclusion, this approach yields rapid, straightforward, and simple DIV1 diagnoses, making it potentially valuable as a reliable tool for the detection and prevention of DIV1, especially where there is a paucity of laboratory equipment. decapod iridescent virus 1 recombinase polymerase amplification <i<Penaeus vannamei</i< white leg shrimp Microbiology Yan Wang verfasserin aut Jingjie Hu verfasserin aut Zhenmin Bao verfasserin aut Mengqiang Wang verfasserin aut In Viruses MDPI AG, 2009 14(2022), 12, p 2752 (DE-627)609775871 (DE-600)2516098-9 19994915 nnns volume:14 year:2022 number:12, p 2752 https://doi.org/10.3390/v14122752 kostenfrei https://doaj.org/article/fc8fe03f1b43447d849f801abe2780a3 kostenfrei https://www.mdpi.com/1999-4915/14/12/2752 kostenfrei https://doaj.org/toc/1999-4915 Journal toc kostenfrei GBV_USEFLAG_A SYSFLAG_A GBV_DOAJ GBV_ILN_11 GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_39 GBV_ILN_40 GBV_ILN_60 GBV_ILN_62 GBV_ILN_63 GBV_ILN_65 GBV_ILN_69 GBV_ILN_70 GBV_ILN_73 GBV_ILN_74 GBV_ILN_95 GBV_ILN_105 GBV_ILN_110 GBV_ILN_151 GBV_ILN_161 GBV_ILN_170 GBV_ILN_206 GBV_ILN_213 GBV_ILN_224 GBV_ILN_230 GBV_ILN_285 GBV_ILN_293 GBV_ILN_370 GBV_ILN_602 GBV_ILN_2005 GBV_ILN_2009 GBV_ILN_2011 GBV_ILN_2014 GBV_ILN_2055 GBV_ILN_2111 GBV_ILN_4012 GBV_ILN_4037 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4249 GBV_ILN_4305 GBV_ILN_4306 GBV_ILN_4307 GBV_ILN_4313 GBV_ILN_4322 GBV_ILN_4323 GBV_ILN_4324 GBV_ILN_4325 GBV_ILN_4338 GBV_ILN_4367 GBV_ILN_4700 AR 14 2022 12, p 2752 |
allfields_unstemmed |
10.3390/v14122752 doi (DE-627)DOAJ082954917 (DE-599)DOAJfc8fe03f1b43447d849f801abe2780a3 DE-627 ger DE-627 rakwb eng QR1-502 Yajin Xu verfasserin aut Development and Visualization Improvement for the Rapid Detection of Decapod Iridescent Virus 1 (DIV1) in <i<Penaeus vannamei</i< Based on an Isothermal Recombinase Polymerase Amplification Assay 2022 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier Viral diseases have seriously restricted the healthy development of aquaculture, and decapod iridescent virus 1 (DIV1) has led to heavy losses in the global shrimp aquaculture industry. Due to the lack of effective treatment, early detection and regular monitoring are the most effective ways to avoid infection with DIV1. In this study, a novel real-time quantitative recombinase polymerase amplification (qRPA) assay and its instrument-free visualization improvement were described for the rapid detection of DIV1. Optimum primer pairs, suitable reaction temperatures, and probe concentrations of a DIV1-qRPA assay were screened to determine optimal reaction conditions. Then, its ability to detect DIV1 was evaluated and compared with real-time quantitative polymerase chain reactions (qPCRs). The sensitivity tests demonstrated that the limit of detection (LOD) of the DIV1-qRPA assay was 1.0 copies μL<sup<−1</sup<. Additionally, the presentation of the detection results was improved with SYBR Green I, and the LOD of the DIV1-RPA-SYBR Green I assay was 1.0 × 10<sup<3</sup< copies μL<sup<−1</sup<. Both the DIV1-qRPA and DIV1-RPA-SYBR Green I assays could be performed at 42 °C within 20 min and without cross-reactivity with the following: white spot syndrome virus (WSSV), <i<Vibrio parahaemolyticus</i< associated with acute hepatopancreatic necrosis disease (<i<V<sub<p</sub<</i<<sub<AHPND</sub<), <i<Enterocytozoon hepatopenaei</i< (EHP), and infectious hypodermal and hematopoietic necrosis virus (IHHNV). In conclusion, this approach yields rapid, straightforward, and simple DIV1 diagnoses, making it potentially valuable as a reliable tool for the detection and prevention of DIV1, especially where there is a paucity of laboratory equipment. decapod iridescent virus 1 recombinase polymerase amplification <i<Penaeus vannamei</i< white leg shrimp Microbiology Yan Wang verfasserin aut Jingjie Hu verfasserin aut Zhenmin Bao verfasserin aut Mengqiang Wang verfasserin aut In Viruses MDPI AG, 2009 14(2022), 12, p 2752 (DE-627)609775871 (DE-600)2516098-9 19994915 nnns volume:14 year:2022 number:12, p 2752 https://doi.org/10.3390/v14122752 kostenfrei https://doaj.org/article/fc8fe03f1b43447d849f801abe2780a3 kostenfrei https://www.mdpi.com/1999-4915/14/12/2752 kostenfrei https://doaj.org/toc/1999-4915 Journal toc kostenfrei GBV_USEFLAG_A SYSFLAG_A GBV_DOAJ GBV_ILN_11 GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_39 GBV_ILN_40 GBV_ILN_60 GBV_ILN_62 GBV_ILN_63 GBV_ILN_65 GBV_ILN_69 GBV_ILN_70 GBV_ILN_73 GBV_ILN_74 GBV_ILN_95 GBV_ILN_105 GBV_ILN_110 GBV_ILN_151 GBV_ILN_161 GBV_ILN_170 GBV_ILN_206 GBV_ILN_213 GBV_ILN_224 GBV_ILN_230 GBV_ILN_285 GBV_ILN_293 GBV_ILN_370 GBV_ILN_602 GBV_ILN_2005 GBV_ILN_2009 GBV_ILN_2011 GBV_ILN_2014 GBV_ILN_2055 GBV_ILN_2111 GBV_ILN_4012 GBV_ILN_4037 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4249 GBV_ILN_4305 GBV_ILN_4306 GBV_ILN_4307 GBV_ILN_4313 GBV_ILN_4322 GBV_ILN_4323 GBV_ILN_4324 GBV_ILN_4325 GBV_ILN_4338 GBV_ILN_4367 GBV_ILN_4700 AR 14 2022 12, p 2752 |
allfieldsGer |
10.3390/v14122752 doi (DE-627)DOAJ082954917 (DE-599)DOAJfc8fe03f1b43447d849f801abe2780a3 DE-627 ger DE-627 rakwb eng QR1-502 Yajin Xu verfasserin aut Development and Visualization Improvement for the Rapid Detection of Decapod Iridescent Virus 1 (DIV1) in <i<Penaeus vannamei</i< Based on an Isothermal Recombinase Polymerase Amplification Assay 2022 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier Viral diseases have seriously restricted the healthy development of aquaculture, and decapod iridescent virus 1 (DIV1) has led to heavy losses in the global shrimp aquaculture industry. Due to the lack of effective treatment, early detection and regular monitoring are the most effective ways to avoid infection with DIV1. In this study, a novel real-time quantitative recombinase polymerase amplification (qRPA) assay and its instrument-free visualization improvement were described for the rapid detection of DIV1. Optimum primer pairs, suitable reaction temperatures, and probe concentrations of a DIV1-qRPA assay were screened to determine optimal reaction conditions. Then, its ability to detect DIV1 was evaluated and compared with real-time quantitative polymerase chain reactions (qPCRs). The sensitivity tests demonstrated that the limit of detection (LOD) of the DIV1-qRPA assay was 1.0 copies μL<sup<−1</sup<. Additionally, the presentation of the detection results was improved with SYBR Green I, and the LOD of the DIV1-RPA-SYBR Green I assay was 1.0 × 10<sup<3</sup< copies μL<sup<−1</sup<. Both the DIV1-qRPA and DIV1-RPA-SYBR Green I assays could be performed at 42 °C within 20 min and without cross-reactivity with the following: white spot syndrome virus (WSSV), <i<Vibrio parahaemolyticus</i< associated with acute hepatopancreatic necrosis disease (<i<V<sub<p</sub<</i<<sub<AHPND</sub<), <i<Enterocytozoon hepatopenaei</i< (EHP), and infectious hypodermal and hematopoietic necrosis virus (IHHNV). In conclusion, this approach yields rapid, straightforward, and simple DIV1 diagnoses, making it potentially valuable as a reliable tool for the detection and prevention of DIV1, especially where there is a paucity of laboratory equipment. decapod iridescent virus 1 recombinase polymerase amplification <i<Penaeus vannamei</i< white leg shrimp Microbiology Yan Wang verfasserin aut Jingjie Hu verfasserin aut Zhenmin Bao verfasserin aut Mengqiang Wang verfasserin aut In Viruses MDPI AG, 2009 14(2022), 12, p 2752 (DE-627)609775871 (DE-600)2516098-9 19994915 nnns volume:14 year:2022 number:12, p 2752 https://doi.org/10.3390/v14122752 kostenfrei https://doaj.org/article/fc8fe03f1b43447d849f801abe2780a3 kostenfrei https://www.mdpi.com/1999-4915/14/12/2752 kostenfrei https://doaj.org/toc/1999-4915 Journal toc kostenfrei GBV_USEFLAG_A SYSFLAG_A GBV_DOAJ GBV_ILN_11 GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_39 GBV_ILN_40 GBV_ILN_60 GBV_ILN_62 GBV_ILN_63 GBV_ILN_65 GBV_ILN_69 GBV_ILN_70 GBV_ILN_73 GBV_ILN_74 GBV_ILN_95 GBV_ILN_105 GBV_ILN_110 GBV_ILN_151 GBV_ILN_161 GBV_ILN_170 GBV_ILN_206 GBV_ILN_213 GBV_ILN_224 GBV_ILN_230 GBV_ILN_285 GBV_ILN_293 GBV_ILN_370 GBV_ILN_602 GBV_ILN_2005 GBV_ILN_2009 GBV_ILN_2011 GBV_ILN_2014 GBV_ILN_2055 GBV_ILN_2111 GBV_ILN_4012 GBV_ILN_4037 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4249 GBV_ILN_4305 GBV_ILN_4306 GBV_ILN_4307 GBV_ILN_4313 GBV_ILN_4322 GBV_ILN_4323 GBV_ILN_4324 GBV_ILN_4325 GBV_ILN_4338 GBV_ILN_4367 GBV_ILN_4700 AR 14 2022 12, p 2752 |
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10.3390/v14122752 doi (DE-627)DOAJ082954917 (DE-599)DOAJfc8fe03f1b43447d849f801abe2780a3 DE-627 ger DE-627 rakwb eng QR1-502 Yajin Xu verfasserin aut Development and Visualization Improvement for the Rapid Detection of Decapod Iridescent Virus 1 (DIV1) in <i<Penaeus vannamei</i< Based on an Isothermal Recombinase Polymerase Amplification Assay 2022 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier Viral diseases have seriously restricted the healthy development of aquaculture, and decapod iridescent virus 1 (DIV1) has led to heavy losses in the global shrimp aquaculture industry. Due to the lack of effective treatment, early detection and regular monitoring are the most effective ways to avoid infection with DIV1. In this study, a novel real-time quantitative recombinase polymerase amplification (qRPA) assay and its instrument-free visualization improvement were described for the rapid detection of DIV1. Optimum primer pairs, suitable reaction temperatures, and probe concentrations of a DIV1-qRPA assay were screened to determine optimal reaction conditions. Then, its ability to detect DIV1 was evaluated and compared with real-time quantitative polymerase chain reactions (qPCRs). The sensitivity tests demonstrated that the limit of detection (LOD) of the DIV1-qRPA assay was 1.0 copies μL<sup<−1</sup<. Additionally, the presentation of the detection results was improved with SYBR Green I, and the LOD of the DIV1-RPA-SYBR Green I assay was 1.0 × 10<sup<3</sup< copies μL<sup<−1</sup<. Both the DIV1-qRPA and DIV1-RPA-SYBR Green I assays could be performed at 42 °C within 20 min and without cross-reactivity with the following: white spot syndrome virus (WSSV), <i<Vibrio parahaemolyticus</i< associated with acute hepatopancreatic necrosis disease (<i<V<sub<p</sub<</i<<sub<AHPND</sub<), <i<Enterocytozoon hepatopenaei</i< (EHP), and infectious hypodermal and hematopoietic necrosis virus (IHHNV). In conclusion, this approach yields rapid, straightforward, and simple DIV1 diagnoses, making it potentially valuable as a reliable tool for the detection and prevention of DIV1, especially where there is a paucity of laboratory equipment. decapod iridescent virus 1 recombinase polymerase amplification <i<Penaeus vannamei</i< white leg shrimp Microbiology Yan Wang verfasserin aut Jingjie Hu verfasserin aut Zhenmin Bao verfasserin aut Mengqiang Wang verfasserin aut In Viruses MDPI AG, 2009 14(2022), 12, p 2752 (DE-627)609775871 (DE-600)2516098-9 19994915 nnns volume:14 year:2022 number:12, p 2752 https://doi.org/10.3390/v14122752 kostenfrei https://doaj.org/article/fc8fe03f1b43447d849f801abe2780a3 kostenfrei https://www.mdpi.com/1999-4915/14/12/2752 kostenfrei https://doaj.org/toc/1999-4915 Journal toc kostenfrei GBV_USEFLAG_A SYSFLAG_A GBV_DOAJ GBV_ILN_11 GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_39 GBV_ILN_40 GBV_ILN_60 GBV_ILN_62 GBV_ILN_63 GBV_ILN_65 GBV_ILN_69 GBV_ILN_70 GBV_ILN_73 GBV_ILN_74 GBV_ILN_95 GBV_ILN_105 GBV_ILN_110 GBV_ILN_151 GBV_ILN_161 GBV_ILN_170 GBV_ILN_206 GBV_ILN_213 GBV_ILN_224 GBV_ILN_230 GBV_ILN_285 GBV_ILN_293 GBV_ILN_370 GBV_ILN_602 GBV_ILN_2005 GBV_ILN_2009 GBV_ILN_2011 GBV_ILN_2014 GBV_ILN_2055 GBV_ILN_2111 GBV_ILN_4012 GBV_ILN_4037 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4249 GBV_ILN_4305 GBV_ILN_4306 GBV_ILN_4307 GBV_ILN_4313 GBV_ILN_4322 GBV_ILN_4323 GBV_ILN_4324 GBV_ILN_4325 GBV_ILN_4338 GBV_ILN_4367 GBV_ILN_4700 AR 14 2022 12, p 2752 |
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Development and Visualization Improvement for the Rapid Detection of Decapod Iridescent Virus 1 (DIV1) in <i<Penaeus vannamei</i< Based on an Isothermal Recombinase Polymerase Amplification Assay |
abstract |
Viral diseases have seriously restricted the healthy development of aquaculture, and decapod iridescent virus 1 (DIV1) has led to heavy losses in the global shrimp aquaculture industry. Due to the lack of effective treatment, early detection and regular monitoring are the most effective ways to avoid infection with DIV1. In this study, a novel real-time quantitative recombinase polymerase amplification (qRPA) assay and its instrument-free visualization improvement were described for the rapid detection of DIV1. Optimum primer pairs, suitable reaction temperatures, and probe concentrations of a DIV1-qRPA assay were screened to determine optimal reaction conditions. Then, its ability to detect DIV1 was evaluated and compared with real-time quantitative polymerase chain reactions (qPCRs). The sensitivity tests demonstrated that the limit of detection (LOD) of the DIV1-qRPA assay was 1.0 copies μL<sup<−1</sup<. Additionally, the presentation of the detection results was improved with SYBR Green I, and the LOD of the DIV1-RPA-SYBR Green I assay was 1.0 × 10<sup<3</sup< copies μL<sup<−1</sup<. Both the DIV1-qRPA and DIV1-RPA-SYBR Green I assays could be performed at 42 °C within 20 min and without cross-reactivity with the following: white spot syndrome virus (WSSV), <i<Vibrio parahaemolyticus</i< associated with acute hepatopancreatic necrosis disease (<i<V<sub<p</sub<</i<<sub<AHPND</sub<), <i<Enterocytozoon hepatopenaei</i< (EHP), and infectious hypodermal and hematopoietic necrosis virus (IHHNV). In conclusion, this approach yields rapid, straightforward, and simple DIV1 diagnoses, making it potentially valuable as a reliable tool for the detection and prevention of DIV1, especially where there is a paucity of laboratory equipment. |
abstractGer |
Viral diseases have seriously restricted the healthy development of aquaculture, and decapod iridescent virus 1 (DIV1) has led to heavy losses in the global shrimp aquaculture industry. Due to the lack of effective treatment, early detection and regular monitoring are the most effective ways to avoid infection with DIV1. In this study, a novel real-time quantitative recombinase polymerase amplification (qRPA) assay and its instrument-free visualization improvement were described for the rapid detection of DIV1. Optimum primer pairs, suitable reaction temperatures, and probe concentrations of a DIV1-qRPA assay were screened to determine optimal reaction conditions. Then, its ability to detect DIV1 was evaluated and compared with real-time quantitative polymerase chain reactions (qPCRs). The sensitivity tests demonstrated that the limit of detection (LOD) of the DIV1-qRPA assay was 1.0 copies μL<sup<−1</sup<. Additionally, the presentation of the detection results was improved with SYBR Green I, and the LOD of the DIV1-RPA-SYBR Green I assay was 1.0 × 10<sup<3</sup< copies μL<sup<−1</sup<. Both the DIV1-qRPA and DIV1-RPA-SYBR Green I assays could be performed at 42 °C within 20 min and without cross-reactivity with the following: white spot syndrome virus (WSSV), <i<Vibrio parahaemolyticus</i< associated with acute hepatopancreatic necrosis disease (<i<V<sub<p</sub<</i<<sub<AHPND</sub<), <i<Enterocytozoon hepatopenaei</i< (EHP), and infectious hypodermal and hematopoietic necrosis virus (IHHNV). In conclusion, this approach yields rapid, straightforward, and simple DIV1 diagnoses, making it potentially valuable as a reliable tool for the detection and prevention of DIV1, especially where there is a paucity of laboratory equipment. |
abstract_unstemmed |
Viral diseases have seriously restricted the healthy development of aquaculture, and decapod iridescent virus 1 (DIV1) has led to heavy losses in the global shrimp aquaculture industry. Due to the lack of effective treatment, early detection and regular monitoring are the most effective ways to avoid infection with DIV1. In this study, a novel real-time quantitative recombinase polymerase amplification (qRPA) assay and its instrument-free visualization improvement were described for the rapid detection of DIV1. Optimum primer pairs, suitable reaction temperatures, and probe concentrations of a DIV1-qRPA assay were screened to determine optimal reaction conditions. Then, its ability to detect DIV1 was evaluated and compared with real-time quantitative polymerase chain reactions (qPCRs). The sensitivity tests demonstrated that the limit of detection (LOD) of the DIV1-qRPA assay was 1.0 copies μL<sup<−1</sup<. Additionally, the presentation of the detection results was improved with SYBR Green I, and the LOD of the DIV1-RPA-SYBR Green I assay was 1.0 × 10<sup<3</sup< copies μL<sup<−1</sup<. Both the DIV1-qRPA and DIV1-RPA-SYBR Green I assays could be performed at 42 °C within 20 min and without cross-reactivity with the following: white spot syndrome virus (WSSV), <i<Vibrio parahaemolyticus</i< associated with acute hepatopancreatic necrosis disease (<i<V<sub<p</sub<</i<<sub<AHPND</sub<), <i<Enterocytozoon hepatopenaei</i< (EHP), and infectious hypodermal and hematopoietic necrosis virus (IHHNV). In conclusion, this approach yields rapid, straightforward, and simple DIV1 diagnoses, making it potentially valuable as a reliable tool for the detection and prevention of DIV1, especially where there is a paucity of laboratory equipment. |
collection_details |
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container_issue |
12, p 2752 |
title_short |
Development and Visualization Improvement for the Rapid Detection of Decapod Iridescent Virus 1 (DIV1) in <i<Penaeus vannamei</i< Based on an Isothermal Recombinase Polymerase Amplification Assay |
url |
https://doi.org/10.3390/v14122752 https://doaj.org/article/fc8fe03f1b43447d849f801abe2780a3 https://www.mdpi.com/1999-4915/14/12/2752 https://doaj.org/toc/1999-4915 |
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author2 |
Yan Wang Jingjie Hu Zhenmin Bao Mengqiang Wang |
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up_date |
2024-07-03T14:43:50.726Z |
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