基于PCR技术的鹿源性产品成分分子检测研究进展
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摘要
鹿产品多种多样且价格昂贵,其中仅鹿科动物梅花鹿和马鹿的产品被视为正品。由于产品加工过程中其形态结构及理化性质均有不同程度的改变,传统方法难以对产品成分进行准确的定性分析,而分子检测技术反应灵敏、准确性高,被越来越多地应用于鹿源性产品成分的检测中。本文基于mtDNA、SRY基因、microsatellite DNA和SNP在分子检测中的应用,对近年来鹿源性产品成分的分子检测方法进行了综述,以期为后续鹿源性产品成分的分子检测研究提供参考。
Deer products are diverse and expensive, only Cervus nippon and Cervus elaphus are considered as certified product. As the morphological structure and physico-chemical properties of the products are changed to different degrees in the processing process of products,it is difficult for the traditional methods to conduct accurate qualitative analysis of the product ingredients. Molecular detection techniques are sensitive and highly accurate,so they are increasingly applied for the detection of deer-derived ingredients. Based on the application of mtDNA, SRY gene, microsatellite DNA and SNP in molecular detection, the methods for molecular detection of the deer-derived ingredients in products in recent years were reviewed in this paper, so as to provide reference for the further research on molecular detection of the deerderived ingredients in products.
Deer products are diverse and expensive, only Cervus nippon and Cervus elaphus are considered as certified product. As the morphological structure and physico-chemical properties of the products are changed to different degrees in the processing process of products,it is difficult for the traditional methods to conduct accurate qualitative analysis of the product ingredients. Molecular detection techniques are sensitive and highly accurate,so they are increasingly applied for the detection of deer-derived ingredients. Based on the application of mtDNA, SRY gene, microsatellite DNA and SNP in molecular detection, the methods for molecular detection of the deer-derived ingredients in products in recent years were reviewed in this paper, so as to provide reference for the further research on molecular detection of the deerderived ingredients in products.
引文
[1]李欢霞,邢秀梅,杨福合,等.鹿产品检测的研究进展[J]中国畜牧兽医,2012,(5):164-169.
[2]Niedzia kowska m, J drzejewska b, Honnen a c. Molecular biogeography of red deer Cervus elaphus from eastern Europe:Insights from mitochondrial DNA sequences[J].Acta theriologica,2011,56(1):1-12.
[3]Sofronova j k., Ilinsky y y., Orishchenko k e.Detection of mutations in mitochondrial DNA by droplet digital PCR[J].Biochemistry,2016,81(10):1031-1037.
[4]González S, Maldonado j e, Jorge o, et al. Identification of the endangered small red brocket deer(Mazama bororo)using noninvasive genetic techniques(Mammalia; Cervidae)[J].Molecular ecology resources, 2010, 9(3):754-758.
[5]Ewart k m,Frankham g j, Mcewing r, et al. An internationally standardized species identification test for use on suspected seized rhinoceros horn in the illegal wildlife trade[J].Forensic Sci Int Genet, 2018, 32:33-39.
[6]Ramírez j d, Duque m c. Guhl F.Phylogenetic reconstruction based on Cytochrome b(Cytb)gene sequences reveals distinct genotypes within Colombian Trypanosoma cruzi I populations[J].Acta tropica, 2011, 119(1):60-65.
[7]Liu j, Wang c, Fu d,et al. Phylogeography of Nanorana parkeri(Anura:Ranidae)and multiple refugia on the Tibetan Plateau revealed by mitochondrial and nuclear DNA[J]. Scientific Reports, 2015,(5):9857.
[8]谷玉娟,张丽华,傅桂莲.鹿茸线粒体DNA的指纹鉴定研究[J].中国药学杂志,2013,48(3):170-173.
[9]孙景昱,张丽华,傅桂莲,等.聚合酶链式反应偶联单链构象多态性分析(PCR-SSCP)技术用于鹿鞭线粒体DNA指纹的研究[J].中国药学杂志,2014,49(9):721-725.
[10]王自强,吕晶,邵泓,等.利用限制性片段长度多态性技术鉴别梅花鹿骨种属来源的研究[J].中国药师,2017,(5):813-816.
[11]王凤霞,陈媛媛,任贵奇,等.鹿血的PCR-RFLP鉴定研究[J].中草药,2018,49(8):1914-1918.
[12]马伊萨兰,吕明星,唐俊妮,等.用Cytb基因序列分析鉴定肉制品掺假[J].中国食品卫生杂志,2016,28(1)48-51.
[13]冯海永,韩建林.羊肉产品中若干动物源性成分的七重PCR检测技术应用研究[J].中国畜牧兽医,2010,37(9):85-90.
[14]施大卫,王利,娄绘芳,等.青石斑鱼细胞色素C氧化酶亚基Ⅰ(COXⅠ)基因的克隆鉴定及表达分析[J].中国水产科学,2009,16(4):467.
[15]陈利伟,赵志亮,邱德义,等.DNA条形码技术在兰州中川机场鼠类鉴定中的应用[J].中国国境卫生检疫杂志,2018,(2):102-105.
[16]张蓉,刘春生,黄璐琦,等.鹿茸饮片的DNA条形码鉴别研究[J].中国药学杂志,2011,(4):263-266.
[17]崔丽娜,杜鹤,刘新成,等.基于COI条形码序列的鹿茸及其混伪品的DNA分子鉴定[J].吉林中医药, 2012,32(4):384-387.
[18]刘冬,钱齐妮,张红印,等.基于COI条形码的鹿类中药材DNA条形码分子鉴定[J].世界科学技术:中医药现代化,2014,(2):274-278.
[19]高晓晨,刘冬,崔丽娜,等.市售鹿茸饮片及鹿茸粉的DNA条形码鉴定[J].吉林中医药,2016,36(7):706-708.
[20]贾静,石林春,徐志超,等.市售鹿茸粉药材的DNA条形码鉴定[J].药学学报,2015,(10):1356-1361.
[21]Haider n, Nabulsi i. AL-SAFADI B.Identification of meat species by PCR-RFLP of the mitochondrial COI, gene[J].Meat science,2012,90(2):490-493.
[22]王爽,李永波,马超峰,等.DNA条形码COI序列在常见肉类鉴别中的应用研究[J].现代食品科技,2016,(1):188-193.
[23]Wang q, Zhang x, Zhang h y, et al. Identification of 12 animal species meat by T-RFLP on the 12S r RNA gene.[J]. Meat science,2010,85(2):265-269.
[24]Siddappa c m, Saini m, Das a, et al. Sequence characterization of mitochondrial 12S r RNA gene in mouse deer(Moschiola indica)for PCR-RFLP based species identification[J]. Molecular Biology International,2013,(5):783925.
[25]白根本,张林源,刘春生,等.鹿源类中药材DNA序列分析及马鹿和梅花鹿的PCR鉴定[J].中草药, 2006,37(10):1566-1569.
[26]苏莹,邵元臣,张然然,等.利用Y染色体SRY基因对马鹿的遗传多样性进行研究[C]//.中国遗传学会.遗传多样性:前沿与挑战-中国的遗传学研究(2013-2015)--2015中国遗传学会大会论文摘要汇编.北京:中国遗传学会,2015.
[27]黄璐琦,袁媛,蒋超,等.基于COI与SRY序列建立鉴别梅花鹿、马鹿或其杂交鹿的方法[J].中国中药杂志, 2017,(23):4588-4592.
[28]王月月,刘雪雪,董坤哲,等.7种家养动物全基因组微卫星分布的差异研究[J].中国畜牧兽医,2015,42(9):2418-2426.
[29]Jiang l w, Nie h t, Li c, et al. The genetic diversity of wild and cultivated Manila clam(Ruditapes philippinarum)revealed by 29 novel microsatellite markers[J]. Electronic Journal of Biotechnology, 2018,34:17-21.
[30]张相伦,游伟,靳青,等.4个肉牛品种微卫星多态性分析[J].中国畜牧兽医,2018,45(1):162-170.
[31]欧阳依娜,江炎庭,孙利民,等.10个云南地方猪种微卫星遗传多样性分析[J].中国畜牧兽医,2018,45(4):992-1001.
[32]Karsli t, Balcioglu m s. Genetic characterization and population structure of six brown layer pure lines by using microsatellite markers[J].Asian Australasian journal of animal sciences,2018,32(1):49-57.
[33]Rania j, Salwa z, Najib h, et al. Microsatellite polymorphism in Tunisian pomegranates(Punica granatum L.):Cultivar genotyping and identification[J].Biochemical Systematics&Ecology,2012,44(10):27-35.
[34]Shangguan j b, Li z b. Development of novel microsatellite markers for Holothurian scabra(Holothuriidae), Apostichopus japonicas(Stichopodidae)and cross-species testing in other sea cucumbers[J].Chinese Journal of Oceanology and Limnology, 2017,(4):1-9.
[35]张正义,邢秀梅,胡鹏飞,等.微卫星标记及其在动物亲缘关系鉴定中的应用[J].基因组学与应用生物学, 2018,37(4):1406-1412.
[36]Frank e z, Alain c f, Ralph k. Genetic structure and effective population sizes in European red deer(Cervus elaphus)at a continental scale:Insights from microsatellite DNA[J]. The Journal of heredity,2016,107(4):318-326.
[37]Norman a j, Street n r, Spong g. De novo SNP discovery in the Scandinavian brown bear(Ursus arctos)[J].PLoS One,2013,8(11):81012.
[38]Ramos a m, Crooijmans r p m a, Affara n a, et al. Design of a high density SNP genotyping assay in the pig using SNPs identified and characterized by next generation sequencing technology[J].PLoS One,2009,4(8):6524.
[39]Amish s j, Hohenlohe p a, Painter s, et al. RAD sequencing yields a high success rate for westslope cutthroat and rainbow trout species-diagnostic SNP assays[J]. Molecular Ecology Resources,2012,12(4):653-660.
[40]Ba h x, Li z p, Yang y f, et al. Development of diagnostic SNP markers to monitor hybridization between sika deer(Cervus nippon)and wapiti(Cervus elaphus)[J]. G3(Bethesda),2018,8(7):2173-2179.
[41]Ba h, Jia b, Wang g, et al. Genome-wide SNP discovery and analysis of genetic diversity in farmed sika deer(Cervus nippon)in Northeast China using double-digest restriction siteassociated dna sequencing[J]. G3(Bethesda), 2017,7(9):3169-3176.
[42]王淑娟,刘梅芬,闫若潜,等.猪瘟病毒实时荧光定量PCR检测方法的建立与应用[J].中国畜牧兽医, 2017,44(7):2112-2118.
[43]杨帆,石顺利,徐娜,等.牛副流感病毒3型3种基因型多重R T-PCR检测方法的建立[J].中国畜牧兽医,2018,45(1):32-38.
[44]梁琳,逄春华,罗亚坤,等.PCV2和PCV3双重纳米PCR方法的建立及初步应用[J].中国畜牧兽医,2017,(12):57-62.
[45]李健友,李家伟,王超,等.牛副流感3型病毒Nano-PCR、LAMP方法的建立及初步应用[J].中国畜牧兽医, 2017,45(10):2837-2844.
[46]朱云飞,初正云,李洪江.SDS-PAGE凝胶电泳法对不同种鹿茸蛋白质差异化研究[J].辽宁中医药大学学报,2017,(6):48-51.
[2]Niedzia kowska m, J drzejewska b, Honnen a c. Molecular biogeography of red deer Cervus elaphus from eastern Europe:Insights from mitochondrial DNA sequences[J].Acta theriologica,2011,56(1):1-12.
[3]Sofronova j k., Ilinsky y y., Orishchenko k e.Detection of mutations in mitochondrial DNA by droplet digital PCR[J].Biochemistry,2016,81(10):1031-1037.
[4]González S, Maldonado j e, Jorge o, et al. Identification of the endangered small red brocket deer(Mazama bororo)using noninvasive genetic techniques(Mammalia; Cervidae)[J].Molecular ecology resources, 2010, 9(3):754-758.
[5]Ewart k m,Frankham g j, Mcewing r, et al. An internationally standardized species identification test for use on suspected seized rhinoceros horn in the illegal wildlife trade[J].Forensic Sci Int Genet, 2018, 32:33-39.
[6]Ramírez j d, Duque m c. Guhl F.Phylogenetic reconstruction based on Cytochrome b(Cytb)gene sequences reveals distinct genotypes within Colombian Trypanosoma cruzi I populations[J].Acta tropica, 2011, 119(1):60-65.
[7]Liu j, Wang c, Fu d,et al. Phylogeography of Nanorana parkeri(Anura:Ranidae)and multiple refugia on the Tibetan Plateau revealed by mitochondrial and nuclear DNA[J]. Scientific Reports, 2015,(5):9857.
[8]谷玉娟,张丽华,傅桂莲.鹿茸线粒体DNA的指纹鉴定研究[J].中国药学杂志,2013,48(3):170-173.
[9]孙景昱,张丽华,傅桂莲,等.聚合酶链式反应偶联单链构象多态性分析(PCR-SSCP)技术用于鹿鞭线粒体DNA指纹的研究[J].中国药学杂志,2014,49(9):721-725.
[10]王自强,吕晶,邵泓,等.利用限制性片段长度多态性技术鉴别梅花鹿骨种属来源的研究[J].中国药师,2017,(5):813-816.
[11]王凤霞,陈媛媛,任贵奇,等.鹿血的PCR-RFLP鉴定研究[J].中草药,2018,49(8):1914-1918.
[12]马伊萨兰,吕明星,唐俊妮,等.用Cytb基因序列分析鉴定肉制品掺假[J].中国食品卫生杂志,2016,28(1)48-51.
[13]冯海永,韩建林.羊肉产品中若干动物源性成分的七重PCR检测技术应用研究[J].中国畜牧兽医,2010,37(9):85-90.
[14]施大卫,王利,娄绘芳,等.青石斑鱼细胞色素C氧化酶亚基Ⅰ(COXⅠ)基因的克隆鉴定及表达分析[J].中国水产科学,2009,16(4):467.
[15]陈利伟,赵志亮,邱德义,等.DNA条形码技术在兰州中川机场鼠类鉴定中的应用[J].中国国境卫生检疫杂志,2018,(2):102-105.
[16]张蓉,刘春生,黄璐琦,等.鹿茸饮片的DNA条形码鉴别研究[J].中国药学杂志,2011,(4):263-266.
[17]崔丽娜,杜鹤,刘新成,等.基于COI条形码序列的鹿茸及其混伪品的DNA分子鉴定[J].吉林中医药, 2012,32(4):384-387.
[18]刘冬,钱齐妮,张红印,等.基于COI条形码的鹿类中药材DNA条形码分子鉴定[J].世界科学技术:中医药现代化,2014,(2):274-278.
[19]高晓晨,刘冬,崔丽娜,等.市售鹿茸饮片及鹿茸粉的DNA条形码鉴定[J].吉林中医药,2016,36(7):706-708.
[20]贾静,石林春,徐志超,等.市售鹿茸粉药材的DNA条形码鉴定[J].药学学报,2015,(10):1356-1361.
[21]Haider n, Nabulsi i. AL-SAFADI B.Identification of meat species by PCR-RFLP of the mitochondrial COI, gene[J].Meat science,2012,90(2):490-493.
[22]王爽,李永波,马超峰,等.DNA条形码COI序列在常见肉类鉴别中的应用研究[J].现代食品科技,2016,(1):188-193.
[23]Wang q, Zhang x, Zhang h y, et al. Identification of 12 animal species meat by T-RFLP on the 12S r RNA gene.[J]. Meat science,2010,85(2):265-269.
[24]Siddappa c m, Saini m, Das a, et al. Sequence characterization of mitochondrial 12S r RNA gene in mouse deer(Moschiola indica)for PCR-RFLP based species identification[J]. Molecular Biology International,2013,(5):783925.
[25]白根本,张林源,刘春生,等.鹿源类中药材DNA序列分析及马鹿和梅花鹿的PCR鉴定[J].中草药, 2006,37(10):1566-1569.
[26]苏莹,邵元臣,张然然,等.利用Y染色体SRY基因对马鹿的遗传多样性进行研究[C]//.中国遗传学会.遗传多样性:前沿与挑战-中国的遗传学研究(2013-2015)--2015中国遗传学会大会论文摘要汇编.北京:中国遗传学会,2015.
[27]黄璐琦,袁媛,蒋超,等.基于COI与SRY序列建立鉴别梅花鹿、马鹿或其杂交鹿的方法[J].中国中药杂志, 2017,(23):4588-4592.
[28]王月月,刘雪雪,董坤哲,等.7种家养动物全基因组微卫星分布的差异研究[J].中国畜牧兽医,2015,42(9):2418-2426.
[29]Jiang l w, Nie h t, Li c, et al. The genetic diversity of wild and cultivated Manila clam(Ruditapes philippinarum)revealed by 29 novel microsatellite markers[J]. Electronic Journal of Biotechnology, 2018,34:17-21.
[30]张相伦,游伟,靳青,等.4个肉牛品种微卫星多态性分析[J].中国畜牧兽医,2018,45(1):162-170.
[31]欧阳依娜,江炎庭,孙利民,等.10个云南地方猪种微卫星遗传多样性分析[J].中国畜牧兽医,2018,45(4):992-1001.
[32]Karsli t, Balcioglu m s. Genetic characterization and population structure of six brown layer pure lines by using microsatellite markers[J].Asian Australasian journal of animal sciences,2018,32(1):49-57.
[33]Rania j, Salwa z, Najib h, et al. Microsatellite polymorphism in Tunisian pomegranates(Punica granatum L.):Cultivar genotyping and identification[J].Biochemical Systematics&Ecology,2012,44(10):27-35.
[34]Shangguan j b, Li z b. Development of novel microsatellite markers for Holothurian scabra(Holothuriidae), Apostichopus japonicas(Stichopodidae)and cross-species testing in other sea cucumbers[J].Chinese Journal of Oceanology and Limnology, 2017,(4):1-9.
[35]张正义,邢秀梅,胡鹏飞,等.微卫星标记及其在动物亲缘关系鉴定中的应用[J].基因组学与应用生物学, 2018,37(4):1406-1412.
[36]Frank e z, Alain c f, Ralph k. Genetic structure and effective population sizes in European red deer(Cervus elaphus)at a continental scale:Insights from microsatellite DNA[J]. The Journal of heredity,2016,107(4):318-326.
[37]Norman a j, Street n r, Spong g. De novo SNP discovery in the Scandinavian brown bear(Ursus arctos)[J].PLoS One,2013,8(11):81012.
[38]Ramos a m, Crooijmans r p m a, Affara n a, et al. Design of a high density SNP genotyping assay in the pig using SNPs identified and characterized by next generation sequencing technology[J].PLoS One,2009,4(8):6524.
[39]Amish s j, Hohenlohe p a, Painter s, et al. RAD sequencing yields a high success rate for westslope cutthroat and rainbow trout species-diagnostic SNP assays[J]. Molecular Ecology Resources,2012,12(4):653-660.
[40]Ba h x, Li z p, Yang y f, et al. Development of diagnostic SNP markers to monitor hybridization between sika deer(Cervus nippon)and wapiti(Cervus elaphus)[J]. G3(Bethesda),2018,8(7):2173-2179.
[41]Ba h, Jia b, Wang g, et al. Genome-wide SNP discovery and analysis of genetic diversity in farmed sika deer(Cervus nippon)in Northeast China using double-digest restriction siteassociated dna sequencing[J]. G3(Bethesda), 2017,7(9):3169-3176.
[42]王淑娟,刘梅芬,闫若潜,等.猪瘟病毒实时荧光定量PCR检测方法的建立与应用[J].中国畜牧兽医, 2017,44(7):2112-2118.
[43]杨帆,石顺利,徐娜,等.牛副流感病毒3型3种基因型多重R T-PCR检测方法的建立[J].中国畜牧兽医,2018,45(1):32-38.
[44]梁琳,逄春华,罗亚坤,等.PCV2和PCV3双重纳米PCR方法的建立及初步应用[J].中国畜牧兽医,2017,(12):57-62.
[45]李健友,李家伟,王超,等.牛副流感3型病毒Nano-PCR、LAMP方法的建立及初步应用[J].中国畜牧兽医, 2017,45(10):2837-2844.
[46]朱云飞,初正云,李洪江.SDS-PAGE凝胶电泳法对不同种鹿茸蛋白质差异化研究[J].辽宁中医药大学学报,2017,(6):48-51.