实时荧光PCR法鉴别玛咖及其掺假物芜菁
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摘要
为建立玛咖原料及其制品中玛咖及其掺假物芜菁物种鉴别方法,根据defensin基因序列,分别设计了玛咖、芜菁的特异引物探针,建立了两种植物源性成分的实时荧光PCR检测方法。以玛咖、芜菁等33份样品DNA为模板,经实时荧光PCR扩增,结果表明:已建立方法分别能特异扩增玛咖、芜菁成分;灵敏度测试结果表明:本方法检测玛咖、芜菁成分的绝对灵敏度均达0.01 ng/μL,相对灵敏度均达0.1%(w/w),且能准确鉴别市售玛咖制品中的玛咖与芜菁成分。由此说明所建立方法可作为检测玛咖原料及其制品中玛咖与掺假物芜菁成分的特异性鉴别方法。
By the defensin gene,specific primers and probes of maca and rappini were designed,and the real-time fluorescence PCR methods for detection of both botanical components were established.The specificity of both methods were verified by using 33 samples DNA,including maca,rappini,and so on.Sensitivity test results showed that the absolute sensitivity for detecting both components reached 0.01 ng/L,the relative sensitivity was 0.1% (w/w),and the methods could accurately identify maca and rappini components in commercial maca products. Above all,the established methods could be used for the specific identification of maca and its adulterated component rappini in maca raw materials and its products.
By the defensin gene,specific primers and probes of maca and rappini were designed,and the real-time fluorescence PCR methods for detection of both botanical components were established.The specificity of both methods were verified by using 33 samples DNA,including maca,rappini,and so on.Sensitivity test results showed that the absolute sensitivity for detecting both components reached 0.01 ng/L,the relative sensitivity was 0.1% (w/w),and the methods could accurately identify maca and rappini components in commercial maca products. Above all,the established methods could be used for the specific identification of maca and its adulterated component rappini in maca raw materials and its products.
引文
[1]张志鹏,万文婷,谭晓蕾,等.云南玛卡主产区栽培技术与生产现状调查[J].中国现代中药,2015,17(12):1305-1307.
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[5]李学芳,邱斌,蒲星宇.玛卡与混淆品蔓菁的比较鉴别[J].中药材,2015,38(8):1643-1645.
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[7]王元忠,赵艳丽,张霁,等.红外光谱结合统计分析对不同产地玛咖的鉴别分类[J].食品科学,2016,37(4):169-175.
[8]刘兴勇,邵金良,陈兴连,等.基于高效液相色谱指纹图谱的玛咖及其制品真实性识别[J].农业工程学报,2016,32(6):302-307.
[9]党艳婷,苑鹏,夏凯,等.基于气味指纹图谱的玛咖品质快速鉴定方法[J].食品科学,2018,39(6):291-297.
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[11]郭燕华,王德莲,刘冬虹,等.重组聚合酶等温扩增技术快速鉴别玛卡及其制品的研究[J].粮食与油脂,2017,30(7):51-54.
[12]谭晓蕾,易帆,谭芳,等.不同产地玛卡遗传多样性的ISSR分析[J].中国现代中药,2016,18(11):1443-1447.
[13]王江瑞,陈文仿,许莉,等.基于ITS2条形码的玛咖药材鉴别及9种元素的测定[J].药物分析杂志,2017(4):677-687.
[14]Zhao J,Wang M,Khan I A.Detection of adulteration in dietary supplements:Maca(Lepidium meyenii)case study by NMRapproach[J].Planta Medica,2015,81(11):12.
[15]Alkahtani H A,Ismail E A,Asif A M.Pork detection in binary meat mixtures and some commercial food products using conventional and real-time PCR techniques[J].Food Chemistry,2017,219:54-60.
[16]Meira L,Costa J,Villa C,et al.EvaGreen real-time PCR to determine horse meat adulteration in processed foods[J].LWT-Food Science and Technology,2017,75:408-416.
[17]Carloni E,Amagliani G,Omiccioli E,et al.Validation and application of a quantitative real-time PCR assay to detect common wheat adulteration of durum wheat for pasta production[J].Food Chemistry,2016,224:86-91.
[18]韩建勋,吴亚君,王斌,等.实时荧光PCR法快速检测食用淀粉中的木薯成分[J].检验检疫学刊,2015(6):4-8.
[19]nirc M,Fekete T,L’ubomír Bele,et al.Detection of ovine milk adulteration using Taqman real-time PCR assay[J].Potravinarstvo,2017,11(1):338-343.
[20]SN/T 3729.6-2013出口食品及饮料中常见水果品种的鉴定方法,第6部分:苹果成分检测,实时荧光PCR法[S].
[21]Dobnik D,Spilsberg B,Bogo6alec A,et al.Multiplex quantification of twelve EU authorized GM maize lines with droplet digital PCR[J].Analytical Chemistry,2015,87(16):8218-8226.
[22]Ren J,Deng T,Huang W,et al.A digital PCR method for identifying and quantifying adulteration of meat species in raw and processed food[J].Plos One,2017,12(3):1-17.
[23]Bucher T B,K9ppel R.Duplex digital droplet PCR for the determination of non-Basmati rice in Basmati rice(Oryza sativa)on the base of a deletion in the fragrant gene[J].European Food Research&Technology,2016,242(6):927-934.
[2]ValentováK,Frcˇek J.Yacon(Smallanthus sonchifolius)and Maca(Lepidium meyenii),Traditional andean crops as new functional foods on the European market[J].Chemicke Listy,2001,95(10):594-901.
[3]Zheng B L,He K,Kim C H,et al.Effect of a lipidic extract from Lepidium meyenii on sexual behavior in mice and rats[J].Urology,2000,55(4):598-602.
[4]郭承刚,薛润光,和寿星,等.不同产地玛咖挥发油成分的GC-MS分析[J].贵州农业科学,2016,44(12):129-132.
[5]李学芳,邱斌,蒲星宇.玛卡与混淆品蔓菁的比较鉴别[J].中药材,2015,38(8):1643-1645.
[6]刘宇,陈文仿,王江瑞,等.红外光谱结合化学计量分析对不同产地玛咖的鉴别[J].中药材,2016,39(12):2733-2737.
[7]王元忠,赵艳丽,张霁,等.红外光谱结合统计分析对不同产地玛咖的鉴别分类[J].食品科学,2016,37(4):169-175.
[8]刘兴勇,邵金良,陈兴连,等.基于高效液相色谱指纹图谱的玛咖及其制品真实性识别[J].农业工程学报,2016,32(6):302-307.
[9]党艳婷,苑鹏,夏凯,等.基于气味指纹图谱的玛咖品质快速鉴定方法[J].食品科学,2018,39(6):291-297.
[10]刘冬虹,王德莲,郭燕华,等.玛卡及其制品的PCR方法鉴别研究[J].食品工业科技,2017,38(1):319-333.
[11]郭燕华,王德莲,刘冬虹,等.重组聚合酶等温扩增技术快速鉴别玛卡及其制品的研究[J].粮食与油脂,2017,30(7):51-54.
[12]谭晓蕾,易帆,谭芳,等.不同产地玛卡遗传多样性的ISSR分析[J].中国现代中药,2016,18(11):1443-1447.
[13]王江瑞,陈文仿,许莉,等.基于ITS2条形码的玛咖药材鉴别及9种元素的测定[J].药物分析杂志,2017(4):677-687.
[14]Zhao J,Wang M,Khan I A.Detection of adulteration in dietary supplements:Maca(Lepidium meyenii)case study by NMRapproach[J].Planta Medica,2015,81(11):12.
[15]Alkahtani H A,Ismail E A,Asif A M.Pork detection in binary meat mixtures and some commercial food products using conventional and real-time PCR techniques[J].Food Chemistry,2017,219:54-60.
[16]Meira L,Costa J,Villa C,et al.EvaGreen real-time PCR to determine horse meat adulteration in processed foods[J].LWT-Food Science and Technology,2017,75:408-416.
[17]Carloni E,Amagliani G,Omiccioli E,et al.Validation and application of a quantitative real-time PCR assay to detect common wheat adulteration of durum wheat for pasta production[J].Food Chemistry,2016,224:86-91.
[18]韩建勋,吴亚君,王斌,等.实时荧光PCR法快速检测食用淀粉中的木薯成分[J].检验检疫学刊,2015(6):4-8.
[19]nirc M,Fekete T,L’ubomír Bele,et al.Detection of ovine milk adulteration using Taqman real-time PCR assay[J].Potravinarstvo,2017,11(1):338-343.
[20]SN/T 3729.6-2013出口食品及饮料中常见水果品种的鉴定方法,第6部分:苹果成分检测,实时荧光PCR法[S].
[21]Dobnik D,Spilsberg B,Bogo6alec A,et al.Multiplex quantification of twelve EU authorized GM maize lines with droplet digital PCR[J].Analytical Chemistry,2015,87(16):8218-8226.
[22]Ren J,Deng T,Huang W,et al.A digital PCR method for identifying and quantifying adulteration of meat species in raw and processed food[J].Plos One,2017,12(3):1-17.
[23]Bucher T B,K9ppel R.Duplex digital droplet PCR for the determination of non-Basmati rice in Basmati rice(Oryza sativa)on the base of a deletion in the fragrant gene[J].European Food Research&Technology,2016,242(6):927-934.
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