基于性状鉴别和特异性PCR技术对膨腹海马的鉴别研究
投稿时间:2023-12-04     点此下载全文
引用本文:王琳,魏霞,焦阳,汪冰,解盈盈,尹雪,石丽,陈娟,孙仟,刘丽,张加余,林永强.基于性状鉴别和特异性PCR技术对膨腹海马的鉴别研究[J].中国现代中药,2024,26(9):1470-1477
DOI:10.13313/j.issn.1673-4890.20231204002
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作者中文名作者英文名单位中文名单位英文名E-Mail
王琳 WANG Lin 山东中医药大学 药学院,山东 济南 250355
山东省食品药品检验研究院 国家药品监督管理局胶类产品质量评价重点实验室/山东省中药标准创新与质量评价工程实验室/中药配方颗粒共性技术山东省工程研究中心,山东 济南 250101
School of Pharmacy, Shandong University of Traditional Chinese Medicine, Jinan 250355, China
Shandong Institute for Food and Drug Control/NMPA Key Laboratory for Quality Evaluation of Gelatin Products/Shandong Engineering Laboratory for Standard Innovation and Quality Evaluation of TCM/Shandong Engineering Research Center for Generic Technologies of Traditional Chinese Medicine Formula Granules, Jinan 250101, China
 
魏霞 WEI Xia 山东省食品药品检验研究院 国家药品监督管理局胶类产品质量评价重点实验室/山东省中药标准创新与质量评价工程实验室/中药配方颗粒共性技术山东省工程研究中心,山东 济南 250101 Shandong Institute for Food and Drug Control/NMPA Key Laboratory for Quality Evaluation of Gelatin Products/Shandong Engineering Laboratory for Standard Innovation and Quality Evaluation of TCM/Shandong Engineering Research Center for Generic Technologies of Traditional Chinese Medicine Formula Granules, Jinan 250101, China  
焦阳 JIAO Yang 山东省食品药品检验研究院 国家药品监督管理局胶类产品质量评价重点实验室/山东省中药标准创新与质量评价工程实验室/中药配方颗粒共性技术山东省工程研究中心,山东 济南 250101 Shandong Institute for Food and Drug Control/NMPA Key Laboratory for Quality Evaluation of Gelatin Products/Shandong Engineering Laboratory for Standard Innovation and Quality Evaluation of TCM/Shandong Engineering Research Center for Generic Technologies of Traditional Chinese Medicine Formula Granules, Jinan 250101, China  
汪冰 WANG Bing 山东省食品药品检验研究院 国家药品监督管理局胶类产品质量评价重点实验室/山东省中药标准创新与质量评价工程实验室/中药配方颗粒共性技术山东省工程研究中心,山东 济南 250101 Shandong Institute for Food and Drug Control/NMPA Key Laboratory for Quality Evaluation of Gelatin Products/Shandong Engineering Laboratory for Standard Innovation and Quality Evaluation of TCM/Shandong Engineering Research Center for Generic Technologies of Traditional Chinese Medicine Formula Granules, Jinan 250101, China  
解盈盈 XIE Ying-ying 山东省食品药品检验研究院 国家药品监督管理局胶类产品质量评价重点实验室/山东省中药标准创新与质量评价工程实验室/中药配方颗粒共性技术山东省工程研究中心,山东 济南 250101 Shandong Institute for Food and Drug Control/NMPA Key Laboratory for Quality Evaluation of Gelatin Products/Shandong Engineering Laboratory for Standard Innovation and Quality Evaluation of TCM/Shandong Engineering Research Center for Generic Technologies of Traditional Chinese Medicine Formula Granules, Jinan 250101, China  
尹雪 YIN Xue 山东省食品药品检验研究院 国家药品监督管理局胶类产品质量评价重点实验室/山东省中药标准创新与质量评价工程实验室/中药配方颗粒共性技术山东省工程研究中心,山东 济南 250101 Shandong Institute for Food and Drug Control/NMPA Key Laboratory for Quality Evaluation of Gelatin Products/Shandong Engineering Laboratory for Standard Innovation and Quality Evaluation of TCM/Shandong Engineering Research Center for Generic Technologies of Traditional Chinese Medicine Formula Granules, Jinan 250101, China  
石丽 SHI Li 山东省食品药品检验研究院 国家药品监督管理局胶类产品质量评价重点实验室/山东省中药标准创新与质量评价工程实验室/中药配方颗粒共性技术山东省工程研究中心,山东 济南 250101 Shandong Institute for Food and Drug Control/NMPA Key Laboratory for Quality Evaluation of Gelatin Products/Shandong Engineering Laboratory for Standard Innovation and Quality Evaluation of TCM/Shandong Engineering Research Center for Generic Technologies of Traditional Chinese Medicine Formula Granules, Jinan 250101, China  
陈娟 CHEN Juan 山东中医药大学 药学院,山东 济南 250355
山东省食品药品检验研究院 国家药品监督管理局胶类产品质量评价重点实验室/山东省中药标准创新与质量评价工程实验室/中药配方颗粒共性技术山东省工程研究中心,山东 济南 250101
School of Pharmacy, Shandong University of Traditional Chinese Medicine, Jinan 250355, China
Shandong Institute for Food and Drug Control/NMPA Key Laboratory for Quality Evaluation of Gelatin Products/Shandong Engineering Laboratory for Standard Innovation and Quality Evaluation of TCM/Shandong Engineering Research Center for Generic Technologies of Traditional Chinese Medicine Formula Granules, Jinan 250101, China
 
孙仟 SUN Qian 山东省食品药品检验研究院 国家药品监督管理局胶类产品质量评价重点实验室/山东省中药标准创新与质量评价工程实验室/中药配方颗粒共性技术山东省工程研究中心,山东 济南 250101 Shandong Institute for Food and Drug Control/NMPA Key Laboratory for Quality Evaluation of Gelatin Products/Shandong Engineering Laboratory for Standard Innovation and Quality Evaluation of TCM/Shandong Engineering Research Center for Generic Technologies of Traditional Chinese Medicine Formula Granules, Jinan 250101, China  
刘丽 LIU Li 山东中医药大学 药学院,山东 济南 250355
山东省食品药品检验研究院 国家药品监督管理局胶类产品质量评价重点实验室/山东省中药标准创新与质量评价工程实验室/中药配方颗粒共性技术山东省工程研究中心,山东 济南 250101
School of Pharmacy, Shandong University of Traditional Chinese Medicine, Jinan 250355, China
Shandong Institute for Food and Drug Control/NMPA Key Laboratory for Quality Evaluation of Gelatin Products/Shandong Engineering Laboratory for Standard Innovation and Quality Evaluation of TCM/Shandong Engineering Research Center for Generic Technologies of Traditional Chinese Medicine Formula Granules, Jinan 250101, China
 
张加余* ZHANG Jia-yu 山东中医药大学 药学院,山东 济南 250355
滨州医学院 中医学院,山东 烟台 264003
School of Pharmacy, Shandong University of Traditional Chinese Medicine, Jinan 250355, China
School of Traditional Chinese Medicine, Binzhou Medical University, Yantai 264003, China
 
林永强* LIN Yong-qiang 山东省食品药品检验研究院 国家药品监督管理局胶类产品质量评价重点实验室/山东省中药标准创新与质量评价工程实验室/中药配方颗粒共性技术山东省工程研究中心,山东 济南 250101 Shandong Institute for Food and Drug Control/NMPA Key Laboratory for Quality Evaluation of Gelatin Products/Shandong Engineering Laboratory for Standard Innovation and Quality Evaluation of TCM/Shandong Engineering Research Center for Generic Technologies of Traditional Chinese Medicine Formula Granules, Jinan 250101, China  
基金项目:国家重点研发计划项目(2023YFC3504102);山东省重点研发计划(重大科技创新工程)项目(2021CXGC010511);泉城产业领军人才支持计划创新团队项目
中文摘要:目的 采用性状鉴别和聚合酶链式反应(PCR)技术对我国主要的海马养殖品膨腹海马Hippocampus abdominalis Leeson进行研究,为该品种鉴别技术开发提供数据支持。方法 通过查阅文献资料并结合样品统计观察,总结膨腹海马的性状鉴别特征;根据膨腹海马及其他海马的细胞色素C氧化酶亚基Ⅰ(COⅠ)基因序列,筛选出膨腹海马的特异性单核苷酸多态性(SNP)位点,设计专属膨腹海马的引物PFHM-Primer-F和PFHM-Primer-R。通过优化PCR体系反应条件,考察最佳的PCR反应体系。结果 膨腹海马的核心性状特征主要包括有较多的体环、尾环和背鳍条数,背鳍覆盖4个体环+1个尾环;眼周和鳃盖部具有明显的大而黑的斑点等。基于COⅠ序列展开的PCR鉴别方法研究表明,当退火温度为65 ℃,循环30次时,膨腹海马样品在100~250 bp呈现单一明亮条带,而其他品种均无此条带。结论 建立的性状鉴别和特异性PCR鉴别方法可快速鉴别膨腹海马,为我国海马新资源的鉴别和后续开发提供参考。
中文关键词:膨腹海马  细胞色素C氧化酶亚基Ⅰ  特异性聚合酶链式反应  性状鉴别
 
Identification of Hippocampus abdominalis Based on the Combination of the Morphological Characteristics and Specific PCR
Abstract:Objective Morphological characteristics and polymerase chain reaction (PCR) techniques were used to investigate Hippocampus abdominalis Leeson, the main cultivated species of seahorse in China, to provide data support for the development of identification techniques for this species.Methods Through literature review and statistical observation of samples, morphological identification characteristics of H. abdominalis were summarized. According to the cytochrome C oxidase subunit Ⅰ (COⅠ) gene sequences of the H. abdominalis and other seahorses, specific single nucleotide polymorphism (SNP) sites were screened, and the specific primers PFHM-Primer-F and PFHM-Primer-R were designed. By optimizing the reaction conditions of the PCR system, the optimal PCR reaction system was determined.Results The core morphological characteristics of the H. abdominalis mainly include more body rings, tail rings, and dorsal fins, with the dorsal fin covering 4 body rings +1 tail ring, and noticeable large and black spots around periocular and operculum areas, etc. The PCR identification method based on the COⅠ sequence showed that when the annealing temperature was 65 ℃ and cycled 30 times, H. abdominalis samples displayed a single bright band in the 100-250 bp, while other species showed no such band.Conclusion The morphological identification and specific PCR identification methods established in this study can quickly identify H. abdominalis, providing a reference for the identification and subsequent development of new seahorse resources in China.
keywords:Hippocampus abdominalis Leeson  COⅠ  specific PCR  character identification
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