SYBR GreenⅠReal-Time Fluorescence Quantitative PCR for Parasitic Boveria labialis Identification from the Sea Cucumber Apostichopus japonicus
Boveria disease affects the aquaculture efficiency of sea cucumbers. Boveria labialis causes Boveria disease in Apostichopus japonicus. Owing to the lack of rapid and accurate detection methods, analyzing the transmission route of the pathogen remains complicated. Based on the partial mitochondrial...
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Science Press, PR China
2025-06-01
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| Series: | Progress in Fishery Sciences |
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| Online Access: | http://journal.yykxjz.cn/yykxjz/ch/reader/view_abstract.aspx?file_no=20240322002 |
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| author | Wenyue HAO Jinjin WANG Jianlong GE Bin LI Yingeng WANG Meijie LIAO Xiaojun RONG Hongjing ZHAO Minqi JIANG Wenguang ZHAO Licheng NIU Jiao PAN |
| author_facet | Wenyue HAO Jinjin WANG Jianlong GE Bin LI Yingeng WANG Meijie LIAO Xiaojun RONG Hongjing ZHAO Minqi JIANG Wenguang ZHAO Licheng NIU Jiao PAN |
| author_sort | Wenyue HAO |
| collection | DOAJ |
| description | Boveria disease affects the aquaculture efficiency of sea cucumbers. Boveria labialis causes Boveria disease in Apostichopus japonicus. Owing to the lack of rapid and accurate detection methods, analyzing the transmission route of the pathogen remains complicated. Based on the partial mitochondrial genome sequence of B. labialis obtained using sequencing, primers were designed to amplify nad10 and a SYBR GreenⅠreal-time fluorescent quantitative PCR detection method was established. The concentration of B. labialis in different culture areas and different culture modes of A. japonicus was detected and analyzed. The standard curve established by the designed primers had a good linear relationship in the range of the plasmid standard of 4.05×101–4.05×109 copies/µL, with a reliability (R2) of 0.997. The melting curve showed a single peak, without primer dimers or nonspecific amplification. The minimum detection limit of sensitivity test was 40.5 copies/µL. The designed primers showed specific amplification for B. labialis only and exhibited no cross reaction to Uronema sp., chaenea sp., Colpoda sp. and Paramecium sp. In the repeatability test, the homogeneity of Ct values in the intra-assay and inter-assay of each concentration was high, the CV values of intra- and inter-assay tests were 0.32%–0.82% and 0.40%–0.88%, respectively, indicating good stability. This method was used to detect environmental samples and feeds in different culture areas of four kinds of A. japonicus culture modes. Combined with the comparative analysis of microscopic examination results, there was a moderate positive correlation (R=0.563) between the DNA load of B. labialis in sea water and the infection degree of B. labialis in A. japonicus. A high positive correlation (R=0.931) was found between the DNA load of B. labialis in sediment and attachment samples and the infection degree of B. labialis in A. japonicus. Fresh sea mud was an important carrier for B. labialis transmission. The relevant research results provide reference for the rapid detection, transmission route analysis, and B. labialis prevention and control. |
| format | Article |
| id | doaj-art-0cc6eb68b0ce4c9782cd3414e7666731 |
| institution | Kabale University |
| issn | 2095-9869 |
| language | English |
| publishDate | 2025-06-01 |
| publisher | Science Press, PR China |
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| series | Progress in Fishery Sciences |
| spelling | doaj-art-0cc6eb68b0ce4c9782cd3414e76667312025-08-20T03:48:28ZengScience Press, PR ChinaProgress in Fishery Sciences2095-98692025-06-0146318319310.19663/j.issn2095-9869.2024032200220240322002SYBR GreenⅠReal-Time Fluorescence Quantitative PCR for Parasitic Boveria labialis Identification from the Sea Cucumber Apostichopus japonicusWenyue HAO0Jinjin WANG1Jianlong GE2Bin LI3Yingeng WANG4Meijie LIAO5Xiaojun RONG6Hongjing ZHAO7Minqi JIANG8Wenguang ZHAO9Licheng NIU10Jiao PAN11College of Fisheries and Life Science, Shanghai Ocean University, Shanghai 201306, ChinaState Key Laboratory of Mariculture Biobreeding and Sustainable Goods, Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Qingdao 266071, ChinaState Key Laboratory of Mariculture Biobreeding and Sustainable Goods, Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Qingdao 266071, ChinaState Key Laboratory of Mariculture Biobreeding and Sustainable Goods, Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Qingdao 266071, ChinaState Key Laboratory of Mariculture Biobreeding and Sustainable Goods, Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Qingdao 266071, ChinaState Key Laboratory of Mariculture Biobreeding and Sustainable Goods, Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Qingdao 266071, ChinaState Key Laboratory of Mariculture Biobreeding and Sustainable Goods, Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Qingdao 266071, ChinaState Key Laboratory of Mariculture Biobreeding and Sustainable Goods, Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Qingdao 266071, ChinaState Key Laboratory of Mariculture Biobreeding and Sustainable Goods, Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Qingdao 266071, ChinaState Key Laboratory of Mariculture Biobreeding and Sustainable Goods, Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Qingdao 266071, ChinaState Key Laboratory of Mariculture Biobreeding and Sustainable Goods, Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Qingdao 266071, ChinaInstitute of Evolution and Marine Biodiversity, Ocean University of China, Qingdao 266003, ChinaBoveria disease affects the aquaculture efficiency of sea cucumbers. Boveria labialis causes Boveria disease in Apostichopus japonicus. Owing to the lack of rapid and accurate detection methods, analyzing the transmission route of the pathogen remains complicated. Based on the partial mitochondrial genome sequence of B. labialis obtained using sequencing, primers were designed to amplify nad10 and a SYBR GreenⅠreal-time fluorescent quantitative PCR detection method was established. The concentration of B. labialis in different culture areas and different culture modes of A. japonicus was detected and analyzed. The standard curve established by the designed primers had a good linear relationship in the range of the plasmid standard of 4.05×101–4.05×109 copies/µL, with a reliability (R2) of 0.997. The melting curve showed a single peak, without primer dimers or nonspecific amplification. The minimum detection limit of sensitivity test was 40.5 copies/µL. The designed primers showed specific amplification for B. labialis only and exhibited no cross reaction to Uronema sp., chaenea sp., Colpoda sp. and Paramecium sp. In the repeatability test, the homogeneity of Ct values in the intra-assay and inter-assay of each concentration was high, the CV values of intra- and inter-assay tests were 0.32%–0.82% and 0.40%–0.88%, respectively, indicating good stability. This method was used to detect environmental samples and feeds in different culture areas of four kinds of A. japonicus culture modes. Combined with the comparative analysis of microscopic examination results, there was a moderate positive correlation (R=0.563) between the DNA load of B. labialis in sea water and the infection degree of B. labialis in A. japonicus. A high positive correlation (R=0.931) was found between the DNA load of B. labialis in sediment and attachment samples and the infection degree of B. labialis in A. japonicus. Fresh sea mud was an important carrier for B. labialis transmission. The relevant research results provide reference for the rapid detection, transmission route analysis, and B. labialis prevention and control.http://journal.yykxjz.cn/yykxjz/ch/reader/view_abstract.aspx?file_no=20240322002apostichopus japonicusparasitic diseaseboveria labialisreal-time fluorescent quantitative pcr detectiontransmission route |
| spellingShingle | Wenyue HAO Jinjin WANG Jianlong GE Bin LI Yingeng WANG Meijie LIAO Xiaojun RONG Hongjing ZHAO Minqi JIANG Wenguang ZHAO Licheng NIU Jiao PAN SYBR GreenⅠReal-Time Fluorescence Quantitative PCR for Parasitic Boveria labialis Identification from the Sea Cucumber Apostichopus japonicus Progress in Fishery Sciences apostichopus japonicus parasitic disease boveria labialis real-time fluorescent quantitative pcr detection transmission route |
| title | SYBR GreenⅠReal-Time Fluorescence Quantitative PCR for Parasitic Boveria labialis Identification from the Sea Cucumber Apostichopus japonicus |
| title_full | SYBR GreenⅠReal-Time Fluorescence Quantitative PCR for Parasitic Boveria labialis Identification from the Sea Cucumber Apostichopus japonicus |
| title_fullStr | SYBR GreenⅠReal-Time Fluorescence Quantitative PCR for Parasitic Boveria labialis Identification from the Sea Cucumber Apostichopus japonicus |
| title_full_unstemmed | SYBR GreenⅠReal-Time Fluorescence Quantitative PCR for Parasitic Boveria labialis Identification from the Sea Cucumber Apostichopus japonicus |
| title_short | SYBR GreenⅠReal-Time Fluorescence Quantitative PCR for Parasitic Boveria labialis Identification from the Sea Cucumber Apostichopus japonicus |
| title_sort | sybr greenireal time fluorescence quantitative pcr for parasitic boveria labialis identification from the sea cucumber apostichopus japonicus |
| topic | apostichopus japonicus parasitic disease boveria labialis real-time fluorescent quantitative pcr detection transmission route |
| url | http://journal.yykxjz.cn/yykxjz/ch/reader/view_abstract.aspx?file_no=20240322002 |
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