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A droplet digital PCR method for the detection of scale drop disease virus in yellowfin seabream (Acanthopagrus latus).

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Název: A droplet digital PCR method for the detection of scale drop disease virus in yellowfin seabream (Acanthopagrus latus).
Autoři: Bin Yin, Can Mao, Fangzhao Yu, Wangdong Li, Runhong Pan, Wei Feng, Yong Li
Zdroj: Frontiers in Microbiology; 2024, p1-9, 9p
Témata: VIRUS diseases, DETECTION limit, NUCLEIC acids, STANDARD deviations, SPLEEN
Abstrakt: In this study, a ddPCR method for the detection of scale drop disease virus (SDDV) in yellowfin seabream (Acanthopagrus latus) was established based on Real-time fluorescence quantitative PCR detection methods and principles. The reaction conditions were optimized, and the sensitivity, specificity, accuracy, and reproducibility were assessed. The results showed that threshold line position was determined to be 1900 by the ddPCR method; the optimum annealing temperature for SDDV detection by the ddPCR method was 60°C; the limit of detection was 1.4-1.7 copies/µL; the results of specific detection of other common viruses, except for SDDV specific amplification, were all negative; and the relative standard deviation (RSD) for the reproducibility validation was 0.77%. The samples of yellowfin seabream (Acanthopagrus latus) liver, spleen, kidney, heart, intestine, brain, blood, muscle, skin and ascites with three replicates, respectively, were tested using the ddPCR method, and the results were consistent with clinical findings. The ddPCR method established in this study has the advantages of high sensitivity, high specificity, good reproducibility and simple steps for the quantitative detection of SDDV, which could be used for the nucleic acid detection of clinical SDDV samples, and provided a new quantitative method for the diagnosis of yellowfin seabream SDDV in the early stage of pathogenesis. [ABSTRACT FROM AUTHOR]
Copyright of Frontiers in Microbiology is the property of Frontiers Media S.A. and its content may not be copied or emailed to multiple sites without the copyright holder's express written permission. Additionally, content may not be used with any artificial intelligence tools or machine learning technologies. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)
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Items – Name: Title
  Label: Title
  Group: Ti
  Data: A droplet digital PCR method for the detection of scale drop disease virus in yellowfin seabream (Acanthopagrus latus).
– Name: Author
  Label: Authors
  Group: Au
  Data: <searchLink fieldCode="AR" term="%22Bin+Yin%22">Bin Yin</searchLink><br /><searchLink fieldCode="AR" term="%22Can+Mao%22">Can Mao</searchLink><br /><searchLink fieldCode="AR" term="%22Fangzhao+Yu%22">Fangzhao Yu</searchLink><br /><searchLink fieldCode="AR" term="%22Wangdong+Li%22">Wangdong Li</searchLink><br /><searchLink fieldCode="AR" term="%22Runhong+Pan%22">Runhong Pan</searchLink><br /><searchLink fieldCode="AR" term="%22Wei+Feng%22">Wei Feng</searchLink><br /><searchLink fieldCode="AR" term="%22Yong+Li%22">Yong Li</searchLink>
– Name: TitleSource
  Label: Source
  Group: Src
  Data: Frontiers in Microbiology; 2024, p1-9, 9p
– Name: Subject
  Label: Subject Terms
  Group: Su
  Data: <searchLink fieldCode="DE" term="%22VIRUS+diseases%22">VIRUS diseases</searchLink><br /><searchLink fieldCode="DE" term="%22DETECTION+limit%22">DETECTION limit</searchLink><br /><searchLink fieldCode="DE" term="%22NUCLEIC+acids%22">NUCLEIC acids</searchLink><br /><searchLink fieldCode="DE" term="%22STANDARD+deviations%22">STANDARD deviations</searchLink><br /><searchLink fieldCode="DE" term="%22SPLEEN%22">SPLEEN</searchLink>
– Name: Abstract
  Label: Abstract
  Group: Ab
  Data: In this study, a ddPCR method for the detection of scale drop disease virus (SDDV) in yellowfin seabream (Acanthopagrus latus) was established based on Real-time fluorescence quantitative PCR detection methods and principles. The reaction conditions were optimized, and the sensitivity, specificity, accuracy, and reproducibility were assessed. The results showed that threshold line position was determined to be 1900 by the ddPCR method; the optimum annealing temperature for SDDV detection by the ddPCR method was 60°C; the limit of detection was 1.4-1.7 copies/µL; the results of specific detection of other common viruses, except for SDDV specific amplification, were all negative; and the relative standard deviation (RSD) for the reproducibility validation was 0.77%. The samples of yellowfin seabream (Acanthopagrus latus) liver, spleen, kidney, heart, intestine, brain, blood, muscle, skin and ascites with three replicates, respectively, were tested using the ddPCR method, and the results were consistent with clinical findings. The ddPCR method established in this study has the advantages of high sensitivity, high specificity, good reproducibility and simple steps for the quantitative detection of SDDV, which could be used for the nucleic acid detection of clinical SDDV samples, and provided a new quantitative method for the diagnosis of yellowfin seabream SDDV in the early stage of pathogenesis. [ABSTRACT FROM AUTHOR]
– Name: Abstract
  Label:
  Group: Ab
  Data: <i>Copyright of Frontiers in Microbiology is the property of Frontiers Media S.A. and its content may not be copied or emailed to multiple sites without the copyright holder's express written permission. Additionally, content may not be used with any artificial intelligence tools or machine learning technologies. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract.</i> (Copyright applies to all Abstracts.)
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RecordInfo BibRecord:
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      – Type: doi
        Value: 10.3389/fmicb.2024.1444235
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        Text: English
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        Type: general
      – SubjectFull: DETECTION limit
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      – SubjectFull: NUCLEIC acids
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      – SubjectFull: STANDARD deviations
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      – TitleFull: A droplet digital PCR method for the detection of scale drop disease virus in yellowfin seabream (Acanthopagrus latus).
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            NameFull: Bin Yin
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            NameFull: Can Mao
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            NameFull: Wei Feng
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            – D: 01
              M: 11
              Text: 2024
              Type: published
              Y: 2024
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