Illumina MiSeq高通量测序技术研究原料乳及环境中细菌多样性和致病菌的分布情况
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摘要
利用Illumina MiSeq高通量测序方法测定原料乳及其环境中的细菌微生物的多样性及致病菌的分布情况。提取DNA和PCR扩增后测序,主要测定稀释性曲线、OTU相对丰度曲线以及菌群分布图。结果表明:菌群具有多样性,在输乳管路外侧和盛装原料奶的空贮罐中均检测出金黄色葡萄球菌。应严格监控奶牛生活环境及储奶设施的卫生情况,从而保障原料乳质量安全。
The Illumina MiSeq high-throughput sequencing method was used to determine the bacterial diversity of raw milk and its environment as well as the distribution of pathogenic bacteria and diversity of bacterial microorganisms.This approach is mainly to determine the rarefraction curve, the curve analysis of relative abundance and the flora distribution after DNA extraction and PCR amplification. The results showed that the diversity of the flora, and there are Staphylococcus aureus in the outside of lactiferous pipeline and empty tank with raw milk. So we should strictly control the health situation of the cow's living environment and the milk storage facilities, to ensure the safety of raw milk quality.
The Illumina MiSeq high-throughput sequencing method was used to determine the bacterial diversity of raw milk and its environment as well as the distribution of pathogenic bacteria and diversity of bacterial microorganisms.This approach is mainly to determine the rarefraction curve, the curve analysis of relative abundance and the flora distribution after DNA extraction and PCR amplification. The results showed that the diversity of the flora, and there are Staphylococcus aureus in the outside of lactiferous pipeline and empty tank with raw milk. So we should strictly control the health situation of the cow's living environment and the milk storage facilities, to ensure the safety of raw milk quality.
引文
[1]孙超,吕彬,余传奇,等.牛乳的污染及酶法保鲜方法概述[J].中国牛业科学, 2009, 35(2):34-36.
[2]吕琦,赵凤,毕宇涵,等.原料乳中多种致病菌的快速过滤富集及多重PCR检测[J].食品与发酵工业,2008, 34(12):155-159.
[3]曾冰冰,肖凯军,石磊,等. LAMP方法在食品微生物检测中的应用[J].今日药学, 2007, 17(1):22-25.
[4]王绍祥,杨洲祥,孙真,等.高通量测序技术在水环境微生物群落多样性中的应用[J].化学通报,2014, 77(3):196-203.
[5]田地,马欣,李玉娥,等.利用高通量测序对封存CO2泄漏情景下土壤细菌的研究[J].环境科学,2013, 34(10):401-410.
[6] RUTVISUTTINUNT W, CHINNAWIROTPISAN P,SIMASATHIEN S, et al. Simultaneous and complete genome sequencing of influenza A and B with high coverage by Illumina MiSeq Platform[J]. J Virol Methods, 2013, 193(2):394-404.
[7] WILLIAMS S, FOSTER P, LITTEWOOD D. The complete mitochondrial genome of a turbinid vetigastropod from MiSeq Illumina sequencing of genomic DNA and steps towards a resolved gastropod phylogeny[J]. Gene, 2014, 533(1):38-47.
[8]蔡元锋,贾仲君.基于新一代高通量测序的环境微生物转录组学研究进展[J].生物多样性, 2013, 21(4):401-410.
[9]王兴春,杨致荣,王敏,等.高通量测序技术及其应用[J].中国生物工程杂志, 2012, 32(1):109-114.
[10]段曌,肖炜,王永霞,等. 454测序技术在微生物生态学研究中的应用[J].微生物学杂志, 2011, 31(5):76-81.
[11]徐晓宇,刘和. 454测序法在环境微生物生态研究中的应用[J].生物技术通报, 2010(1):73-77.
[12]周林文. MiSeq:新一代个人化测序仪[J].生物技术世界, 2011(5):14-16.
[13]于国萍,陈媛,姚宇秀,等.利用Illumina MiSeq高通量测序技术分析原料乳的菌群分布[J].食品科学, 2018, 39(16):186-191.
[14]娄勇军,肖小春,陈伟鸿,等.基于ITS2序列和特定引物区分铁皮石斛和伪品的PCR鉴别方法[J].中医药导报, 2018, 24(15):49-52.
[15] BATES S T, CLEMENTE J C, FLORES G E, et al. Global biogeography of highly diverse protis tan communities in soil[J]. The ISME Journal, 2013, 7(3):652-659.
[2]吕琦,赵凤,毕宇涵,等.原料乳中多种致病菌的快速过滤富集及多重PCR检测[J].食品与发酵工业,2008, 34(12):155-159.
[3]曾冰冰,肖凯军,石磊,等. LAMP方法在食品微生物检测中的应用[J].今日药学, 2007, 17(1):22-25.
[4]王绍祥,杨洲祥,孙真,等.高通量测序技术在水环境微生物群落多样性中的应用[J].化学通报,2014, 77(3):196-203.
[5]田地,马欣,李玉娥,等.利用高通量测序对封存CO2泄漏情景下土壤细菌的研究[J].环境科学,2013, 34(10):401-410.
[6] RUTVISUTTINUNT W, CHINNAWIROTPISAN P,SIMASATHIEN S, et al. Simultaneous and complete genome sequencing of influenza A and B with high coverage by Illumina MiSeq Platform[J]. J Virol Methods, 2013, 193(2):394-404.
[7] WILLIAMS S, FOSTER P, LITTEWOOD D. The complete mitochondrial genome of a turbinid vetigastropod from MiSeq Illumina sequencing of genomic DNA and steps towards a resolved gastropod phylogeny[J]. Gene, 2014, 533(1):38-47.
[8]蔡元锋,贾仲君.基于新一代高通量测序的环境微生物转录组学研究进展[J].生物多样性, 2013, 21(4):401-410.
[9]王兴春,杨致荣,王敏,等.高通量测序技术及其应用[J].中国生物工程杂志, 2012, 32(1):109-114.
[10]段曌,肖炜,王永霞,等. 454测序技术在微生物生态学研究中的应用[J].微生物学杂志, 2011, 31(5):76-81.
[11]徐晓宇,刘和. 454测序法在环境微生物生态研究中的应用[J].生物技术通报, 2010(1):73-77.
[12]周林文. MiSeq:新一代个人化测序仪[J].生物技术世界, 2011(5):14-16.
[13]于国萍,陈媛,姚宇秀,等.利用Illumina MiSeq高通量测序技术分析原料乳的菌群分布[J].食品科学, 2018, 39(16):186-191.
[14]娄勇军,肖小春,陈伟鸿,等.基于ITS2序列和特定引物区分铁皮石斛和伪品的PCR鉴别方法[J].中医药导报, 2018, 24(15):49-52.
[15] BATES S T, CLEMENTE J C, FLORES G E, et al. Global biogeography of highly diverse protis tan communities in soil[J]. The ISME Journal, 2013, 7(3):652-659.