泰国香米质量安全分子指纹检测技术探讨Research on detection technology of Thai fragrant rice quality and safety of molecular fingerprint 摘要:（摘要内容经过系统自动伪原创处理以避免复制，下载原文正常，内容请直接查看目录。） 本研究采取RAPD、多重荧光PCR一毛细管一SSCP、2DE、MALDI一TOF等古代份子生物学技巧，展开泰国茉莉喷鼻米种类判定和纯度检测、喷鼻米新陈度检测、食源性致病菌高通量疾速判定办法研究，并停止办法尺度化研究，目标是增强我国进出口食物平安保证技巧贮备，为港口年夜米的检验供给先辈的迷信手腕，进步我国年夜米质量和平安的监管程度，掩护花费者好处，增进年夜米进出口商业的正常运转。研究共分三个部门，第一部门是泰国茉莉喷鼻米的种类判定和纯度检测。采取RAPD指纹技巧，经由过程对120条随机引物的挑选，搜集泰国茉莉喷鼻米法定种类KDML105和RD15和具有代表性的非茉莉喷鼻米合计32个水稻种类的指纹图谱数据，依据指纹数据停止种类聚类关系剖析，注解RAPD指纹分类与传统分类根本分歧。为了进步办法的可操作性，将RAPD办法部门转化为SCAR，树立了R2一449 RAPD和R5一1107SCAR组成的泰国茉莉喷鼻米种类判定和纯度检测办法。上述两个份子标志双隐性者为泰国茉莉喷鼻米。经由过程屡次试验证明了该办法对泰国茉莉喷鼻米法定种类KDML105和RD15的分辩才能、稳固性和定量精确性。第二部门为泰国喷鼻米新陈度判定办法研究。针对今朝年夜米新陈度脂肪酸检测尺度办法所存在的一些限制，采取2DGE技巧，对6个分歧年份的泰国喷鼻米样品停止卵白质组剖析，并对重要卵白点停止肽指纹图谱剖析，联合水稻卵白质组数据库，对卵白质停止判定。成果显示卵白质指纹图谱出现与喷鼻米年份分歧的变更纪律，泰语论文网站，碱性卵白先降解，谷卵白是变更最快的卵白。2017年和2017年份样品差别显著，解释该办法具有较高的分辩率，别的因为卵白质指纹信息量年夜，有益于对年夜米蕴藏的生物学进程停止剖析，懂得年夜米品德的变更纪律，为食粮的临盆、贮存和流畅进程中各个环节的质量掌握供给迷信指点。第三部门为年夜米产物中食源性致病菌疾速判定办法研究。针对传统的微生物判定办法耗时辛苦，和惯例PCR检测办法须要对特定微生物设计特定引物的情形，运用多重荧光PCR一CE一SSCP技巧高通量、疾速、主动化的特色，树立16srRNA和gyrB基因的多重荧光PCR系统，经由过程毛细管电泳对扩增片断的构象多态性停止剖析，搜集了25个尺度菌株的指纹数据，泰语毕业论文，采取RAPD剖析办法对菌种停止聚类剖析，成果显示SSCP聚类关系和传统分类根本分歧。办法运用份子内标绝对地位来检测旌旗灯号峰，屡次试验证明了SSCP旌旗灯号峰具有很好的重现性。别的，因为最近几年来新涌现阪崎杆菌日趋遭到存眷，例如国际上屡次报导了从婴幼儿米粉中检出该菌。是以研究针对42株从食物基质分别的阪崎杆菌停止了SSCP剖析，为阪崎杆菌检测办法的进一步优化和该菌的菌群特点供给了参考数据。 Abstract: This research take RAPD, multiplex fluorescence PCR capillary SSCP, 2DE and MALDI a TOF ancient molecular biology techniques, the Thai Jasmine nasal spray rice varieties judgment and purity testing, nasal spray rice freshness detection, foodborne pathogenic bacteria high-throughput rapid way to determine research, and stop the way standardization research, the goal is enhanced China's import and export food safety assurance technical reserves, the port of the eve of the meter test supply advanced science and wrist, progress the eve of the rice quality and safe level of supervision, cover cost benefits, enhance the eve of rice import and export business of the normal operation of the. Research is divided into three parts. The first is the identification of species in Thailand Jasmine nasal spray and purity detection meter. Take RAPD fingerprinting techniques, through a process of 120 random primers were selected to collect Thailand Jasmine spray nasal meters legal category KDML105 and RD15 and with representatives of non Jasmine spray nasal meters total 32 rice variety fingerprint data, according to the fingerprint data types of relational clustering analyse, annotation RAPD fingerprint classification and traditional classification of fundamental differences. In order to improve the way of operation, the RAPD Department way into SCAR, and establish a detection method to judge the R2 449 RAPD and R5 1107SCAR of Thailand jasmine rice varieties and the purity of the nasal spray. The two part is the sign of the double recessive Thailand Jasmine spray nasal meters. Through the process repeatedly test, it is proved that the approach can to differentiate the Thai Jasmine spray nasal meters legal category KDML105 and RD15, stable and quantitative accuracy. The second departments of the Thailand nasal spray rice freshness determination methods research. For current Nianye Rice Freshness fatty acid detection scale method exists some limitations, take 2DGE skills, protein group analysis of six years of disagreement of Thailand nasal spray rice samples, and the egg white to stop peptide mass fingerprinting analysis, combined with rice protein group data base, to protein judge. Results show that protein fingerprint and spray nasal meters year differences change discipline, alkaline protein degradation, valley albumen is change the fastest proteins. There are significant differences between 2017 and 2017 Vintage samples, explaining the approach has higher resolution and other because the protein fingerprint information amounts are big, beneficial to stop on the eve of the rice contains biological processes analysis, understand the eve of the moral m change discipline and provide scientific guidance for the food production, storage and circulation process each link quality to grasp. The third sector is the product of rice food borne pathogens rapid determination methods research. 16S rRNA and gyrB multiplex fluorescent PCR system, and establish the characteristics of CE for traditional microbiological determination method time-consuming hard, and conventional PCR detection methods need to design specific primers of specific microbial and application of multiplex fluorescent PCR SSCP technique for high-throughput, rapid, initiative, through process of capillary electrophoresis of amplified fragment conformation polymorphism analysis, collect the fingerprint data of 25 strains of scale by RAPD analysis method to the strain clustering analysis results, it showed that the fundamental differences in SSCP clustering relationships and traditional classification. Way to use molecular scale absolute position to detect the signal peak, repeated test, it is proved that the SSCP signal peak with a good reproducibility. Other, because in recent years, the emergence of new Enterobacter sakazakii are increasingly being of concern, such as international repeatedly reported detection of the bacteria from rice flour for infants and young children. To research for 42 strains from the food matrix respectively of Enterobacter sakazakii stop the SSCP analysis, to provide reference data for the further optimization of Enterobacter sakazakii detection method and the bacteria flora characteristics. 目录: |
