Application of loop-mediated isothermal amplification (LAMP) for rapid detection of Atlantic cod (Gadus morhua), Pacific cod (Gadus macrocephalus) and haddock (Melanogrammus aeglefinus). (October 2019)

• Record Type: Journal Article
• Authors: Wang, Yi; Feng, Junli; Tian, Xiaolan
• Is Part Of: Molecular and cellular probes. Volume 47(2019)
• Publication Date: 2019-10
• Subjects: Cod -- Loop-mediated isothermal amplification -- Species identification -- Rapid detection; Molecular probes -- Diagnostic use -- Periodicals; Pathology, Cellular -- Technique -- Periodicals; Cell Biology -- Periodicals; Molecular Biology -- Periodicals; Sondes moléculaires -- Utilisation diagnostique -- Périodiques; Cytopathologie -- Technique -- Périodiques; 572
• DOI: 10.1016/j.mcp.2019.07.003
• Languages: English
• ISSNs: 0890-8508

Abstract

Abstract: Codfish is a commercially important species of sea fish and plays an important role in the world fishery. In our study, two loop-mediated isothermal amplification (LAMP) assays (real-time fluorescence LAMP and visual LAMP) were established for the identification of three cod species in Gadidae ( Gadus morhua, Gadus macrocephalus and Melanogrammus aeglefinus ). 12S rDNA gene was used to design primers to distinguish the Gadidae and non- Gadidae species, and the mitochondrial Cytb gene was selected for discrimination of three cod species. After optimization, the 12S rDNA system and species-specific systems performed well, and target cod DNA could be detected in single or mixed samples. In the species-specific systems, the absolute limit of detection (LODa) of three cod species were 285, 37 and 197 pg/μL, and the relative limit of detection (LODr) reached to 1%, 0.1% and 1%, respectively. In the 12S rDNA system, the LODa of three cod species were 28.5, 37 and 19.7 pg/μL, respectively, and the LODr reached to 0.1%. Through the detection of 13 commercial cod products, the LAMP systems can detect cod contents in raw materials and deep-processed products as well. It indicated that the methods developed in this study have strong practicability and can meet the needs of routine testing. Highlights: Two LAMP systems (real-time fluorescence LAMP and visual LAMP) were established for rapid identification of three cod species in Gadidae. In the 12S rDNA system (universal primer for cod species in Gadidae ), the LODa of three cod species were 28.5, 37 and 19.7 pg/μL, respectively, and the LODr reached to 0.1%. The LAMP systems can detect cod contents in raw materials, mixed and processd cod specimens, and detection time was 60 min or less. The developed assays are suitable for rapid and on-site detection of cod species in Gadidae.