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Background <p>Early and specific detection of the invasive hydroid <i>Cordylophora caspia</i> is crucial for management of its ecological and infrastructural impacts. Here we present the development and laboratory validation of a species-specific Loop-Mediated Isothermal Amplification assay (Caspia-LAMP) targeting repetitive genomic sequences unique to <i>C. caspia</i>.</p> Methods and results <p>Draft genome data (unpublished) were screened for high-copy repetitive elements and candidate regions were selected after BLAST searches to avoid homology with non-target taxa. Caspia-LAMP primers were designed and optimized; reactions used Bst DNA Polymerase under isothermal conditions (65&#xa0;°C). Limit of detection assays (serial dilutions from 10 to 10⁻⁸ ng) and minimum reaction time trials (30–120&#xa0;min) were performed. Cross-reactivity tests included DNA from available cnidarians and several invasive bivalves co-occurring with <i>C. caspia</i>. Caspia-LAMP detected <i>C. caspia</i> DNA down to 10⁻⁷ ng and produced detectable results within 30&#xa0;min; no amplification was observed for the non-target DNA samples tested.</p> Conclusions <p>Caspia-LAMP is a rapid, sensitive, and specific laboratory assay for detection of <i>C. caspia</i> DNA and a promising tool for early surveillance and eDNA applications. We acknowledge limitations related to the panel of cnidarians tested and proposed further validation with additional taxa. The method and primer sequences are provided to facilitate independent validation and field adaptation.</p>

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Caspia-LAMP – a preliminary species-specific LAMP-based tool for the molecular detection of Cordylophora caspia (Pallas 1771) (Cnidaria, Hydrozoa)

  • 期刊论文

  • 发表日期：

    2025年12月15日

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  • Rayan Silva de Paula,
  • Júlia Meireles Nogueira,
  • Amanda Maria Siqueira Moreira,
  • Rubens Lima do Monte-Neto,
  • Gabriel da Luz Wallau,
  • Elverson Soares de Melo,
  • Erika Cristina Jorge,
  • Lucília Souza Miranda

摘要

Background

Early and specific detection of the invasive hydroid Cordylophora caspia is crucial for management of its ecological and infrastructural impacts. Here we present the development and laboratory validation of a species-specific Loop-Mediated Isothermal Amplification assay (Caspia-LAMP) targeting repetitive genomic sequences unique to C. caspia.

Methods and results

Draft genome data (unpublished) were screened for high-copy repetitive elements and candidate regions were selected after BLAST searches to avoid homology with non-target taxa. Caspia-LAMP primers were designed and optimized; reactions used Bst DNA Polymerase under isothermal conditions (65 °C). Limit of detection assays (serial dilutions from 10 to 10⁻⁸ ng) and minimum reaction time trials (30–120 min) were performed. Cross-reactivity tests included DNA from available cnidarians and several invasive bivalves co-occurring with C. caspia. Caspia-LAMP detected C. caspia DNA down to 10⁻⁷ ng and produced detectable results within 30 min; no amplification was observed for the non-target DNA samples tested.

Conclusions

Caspia-LAMP is a rapid, sensitive, and specific laboratory assay for detection of C. caspia DNA and a promising tool for early surveillance and eDNA applications. We acknowledge limitations related to the panel of cnidarians tested and proposed further validation with additional taxa. The method and primer sequences are provided to facilitate independent validation and field adaptation.