The aim is to develop a liquid chip technique to detect Taura syndrome virus( TSV) and yellow head disease virus( YHDV) on Penaeus orientalis simultaneously. The CP2 gene of TSV and N gene of YHDV in Gen Bank was anal...The aim is to develop a liquid chip technique to detect Taura syndrome virus( TSV) and yellow head disease virus( YHDV) on Penaeus orientalis simultaneously. The CP2 gene of TSV and N gene of YHDV in Gen Bank was analysed by using the software DNAStar 7. 0 to design the TSV-and YHDV-specific primers. The primers were labeled with biotin and subjected to amination modification. They were then coupled with fluorescence-coded microspheres and then used for hybridization with RT- PCR products of TSV and YHDV. The liquid chip detection technique for detection of TSV and YHDV was established by using BD FACSArray to detect fluorescence signal in the reaction system. This assay system had a high sensitivity to TSV and YHDV,with the detection of limit of 100 pg. Moreover,the assay was specific for the detection of TSV,YHDV and was not susceptible to cross with other viruses,including white spot syndrome virus( WSSV),spring viremia of carp virus( SVCV),infectious haematopoietic necrosis virus( IHNV). In conclusion,the liquid chip assay technique established in this study is highly sensitive and specific to TSV and YHDV detection. Moreover,it provides a novel,convenient and rapid approach for the detection of TSV and YHDV.展开更多
Genotyping platforms,as critical supports for genomics,genetics,and molecular breeding,have been well implemented at national institutions/universities in developed countries and multinational seed companies that poss...Genotyping platforms,as critical supports for genomics,genetics,and molecular breeding,have been well implemented at national institutions/universities in developed countries and multinational seed companies that possess high-throughput,automatic,large-scale,and shared facilities.In this study,we integrated an improved genotyping by target sequencing(GBTS)system with capture-in-solution(liquid chip)technology to develop a multiple single-nucleotide polymorphism(mSNP)approach in which mSNPs can be captured from a single amplicon.From one 40K maize mSNP panel,we developed three types of markers(40K mSNPs,251K SNPs,and 690K haplotypes),and generated multiple panels with various marker densities(1K–40K mSNPs)by sequencing at different depths.Comparative genetic diversity analysis was performed with genic versus intergenic markers and di-allelic SNPs versus non-typical SNPs.Compared with the one-amplicon-one-SNP system,mSNPs and within-mSNP haplotypes are more powerful for genetic diversity detection,linkage disequilibrium decay analysis,and genome-wide association studies.The technologies,protocols,and application scenarios developed for maize in this study will serve as a model for the development of mSNP arrays and highly efficient GBTS systems in animals,plants,and microorganisms.展开更多
本研究根据公开发表的烟草K326基因组和烟草430K SNP固相芯片检测数据,以7份种质两两组合之间每条染色体上20个多态标记为目标,基于多重PCR扩增的精准定位测序分型技术(mGPS,Genotyping by Pinpoint Sequencing of multiplex PCR produc...本研究根据公开发表的烟草K326基因组和烟草430K SNP固相芯片检测数据,以7份种质两两组合之间每条染色体上20个多态标记为目标,基于多重PCR扩增的精准定位测序分型技术(mGPS,Genotyping by Pinpoint Sequencing of multiplex PCR products)开发出烟草1.8K育种液相芯片(YT1.8K.1)。利用该芯片对上述7份种质两两之间杂交的21个杂交组合进行基因分型检测,每个杂交组合之间的平均差异位点数为650个,能同时满足每个组合定向改良筛选高遗传背景回复率单株的需要。利用该芯片对23个烟草品种进行基因型分型检测和聚类分析,聚类分类结果与品种系谱基本吻合;利用该芯片从367个BC2F1群体中筛选出5个背景回复率高于94.96%的单株,高于理论均值87.5%,表明该育种芯片可应用于烟草种质资源聚类分析、定向改良育种的遗传背景筛选。展开更多
目的:通过液相芯片技术评价消积饮联合化疗对晚期结直肠癌患者血液细胞因子表达谱的影响。方法:回顾性分析2018年1月1日至2018年12月31日来自广东省中医院大学城分院肿瘤科的14例符合纳入标准的晚期结直肠癌患者,根据治疗分为化疗组(n=7...目的:通过液相芯片技术评价消积饮联合化疗对晚期结直肠癌患者血液细胞因子表达谱的影响。方法:回顾性分析2018年1月1日至2018年12月31日来自广东省中医院大学城分院肿瘤科的14例符合纳入标准的晚期结直肠癌患者,根据治疗分为化疗组(n=7)和联合治疗组(n=7),化疗组给予5-氟尿嘧啶+亚叶酸钙+奥沙利铂(FOLFOX)治疗,联合治疗组给予消积饮+FOLFOX治疗,评价两组患者治疗6程后疗效,并在每2个疗程后采用液相芯片技术检测患者外周静脉血清中细胞因子表达谱。结果:14例患者共接受了84程治疗,生存分析显示联合治疗组和化疗组中患者的PFS、OS因样本量不足不能进行对比,但联合治疗组较化疗组的PFS及OS均有延长的趋势(PFS:10 vs 6个月,OS:17 vs 12个月);不良反应方面,两组出现白细胞减少、腹泻、恶心、末梢神经炎及脱发等不良反应发生例数相当,但联合治疗组较化疗组不良反应程度略轻。化疗组患者血清中IL-2和脑源性神经营养血液细胞因子(Brain derived neurotrophic blood cytokines,BDNF)的浓度高于联合治疗组(P<0.05)。比较治疗前后不同采血点细胞因子浓度,患者治疗2程后化疗组的IL-2浓度高于联合治疗组(P<0.05)。在不同用药周期,共有19个细胞因子出现联合治疗组高于化疗组的趋势。结论:消积饮联合FOLFOX方案可能是晚期结直肠癌的一种值得探索的治疗方案,液相芯片分析其机制可能与降低患者血清中细胞因子IL-2和BDNF水平有关。展开更多
基金Supported by Science and Technology Project of General Administration of Quality Supervision,Inspection and Quarantine of the People's Republic of China(2012IK018)Special Fund for Scientific Research in the Public Welfare(201210055-4)
文摘The aim is to develop a liquid chip technique to detect Taura syndrome virus( TSV) and yellow head disease virus( YHDV) on Penaeus orientalis simultaneously. The CP2 gene of TSV and N gene of YHDV in Gen Bank was analysed by using the software DNAStar 7. 0 to design the TSV-and YHDV-specific primers. The primers were labeled with biotin and subjected to amination modification. They were then coupled with fluorescence-coded microspheres and then used for hybridization with RT- PCR products of TSV and YHDV. The liquid chip detection technique for detection of TSV and YHDV was established by using BD FACSArray to detect fluorescence signal in the reaction system. This assay system had a high sensitivity to TSV and YHDV,with the detection of limit of 100 pg. Moreover,the assay was specific for the detection of TSV,YHDV and was not susceptible to cross with other viruses,including white spot syndrome virus( WSSV),spring viremia of carp virus( SVCV),infectious haematopoietic necrosis virus( IHNV). In conclusion,the liquid chip assay technique established in this study is highly sensitive and specific to TSV and YHDV detection. Moreover,it provides a novel,convenient and rapid approach for the detection of TSV and YHDV.
基金This research is supported by the National Key Research and Development Program of China(2016YFD0101803 and 2017YFD0101201)the Central Public-interest Scientific Institution Basal Research Fund(Y2020PT20)+4 种基金the Agricultural Science and Technology Innovation Program(ASTIP)of the Chinese Academy of Agricultural Sciences(CAAS)(CAAS-XTCX2016009)the Key Research Area and Development Program of Guangdong Province(2018B020202008)the Shijiazhuang Science and Technology Incubation Program(191540089A)the Hebei Innovation Capability Enhancement Project(19962911D)Research activities at CIMMYT were supported by the Bill and Melinda Gates Foundation and the CGIAR Research Program MAIZE.
文摘Genotyping platforms,as critical supports for genomics,genetics,and molecular breeding,have been well implemented at national institutions/universities in developed countries and multinational seed companies that possess high-throughput,automatic,large-scale,and shared facilities.In this study,we integrated an improved genotyping by target sequencing(GBTS)system with capture-in-solution(liquid chip)technology to develop a multiple single-nucleotide polymorphism(mSNP)approach in which mSNPs can be captured from a single amplicon.From one 40K maize mSNP panel,we developed three types of markers(40K mSNPs,251K SNPs,and 690K haplotypes),and generated multiple panels with various marker densities(1K–40K mSNPs)by sequencing at different depths.Comparative genetic diversity analysis was performed with genic versus intergenic markers and di-allelic SNPs versus non-typical SNPs.Compared with the one-amplicon-one-SNP system,mSNPs and within-mSNP haplotypes are more powerful for genetic diversity detection,linkage disequilibrium decay analysis,and genome-wide association studies.The technologies,protocols,and application scenarios developed for maize in this study will serve as a model for the development of mSNP arrays and highly efficient GBTS systems in animals,plants,and microorganisms.
文摘本研究根据公开发表的烟草K326基因组和烟草430K SNP固相芯片检测数据,以7份种质两两组合之间每条染色体上20个多态标记为目标,基于多重PCR扩增的精准定位测序分型技术(mGPS,Genotyping by Pinpoint Sequencing of multiplex PCR products)开发出烟草1.8K育种液相芯片(YT1.8K.1)。利用该芯片对上述7份种质两两之间杂交的21个杂交组合进行基因分型检测,每个杂交组合之间的平均差异位点数为650个,能同时满足每个组合定向改良筛选高遗传背景回复率单株的需要。利用该芯片对23个烟草品种进行基因型分型检测和聚类分析,聚类分类结果与品种系谱基本吻合;利用该芯片从367个BC2F1群体中筛选出5个背景回复率高于94.96%的单株,高于理论均值87.5%,表明该育种芯片可应用于烟草种质资源聚类分析、定向改良育种的遗传背景筛选。
文摘目的:通过液相芯片技术评价消积饮联合化疗对晚期结直肠癌患者血液细胞因子表达谱的影响。方法:回顾性分析2018年1月1日至2018年12月31日来自广东省中医院大学城分院肿瘤科的14例符合纳入标准的晚期结直肠癌患者,根据治疗分为化疗组(n=7)和联合治疗组(n=7),化疗组给予5-氟尿嘧啶+亚叶酸钙+奥沙利铂(FOLFOX)治疗,联合治疗组给予消积饮+FOLFOX治疗,评价两组患者治疗6程后疗效,并在每2个疗程后采用液相芯片技术检测患者外周静脉血清中细胞因子表达谱。结果:14例患者共接受了84程治疗,生存分析显示联合治疗组和化疗组中患者的PFS、OS因样本量不足不能进行对比,但联合治疗组较化疗组的PFS及OS均有延长的趋势(PFS:10 vs 6个月,OS:17 vs 12个月);不良反应方面,两组出现白细胞减少、腹泻、恶心、末梢神经炎及脱发等不良反应发生例数相当,但联合治疗组较化疗组不良反应程度略轻。化疗组患者血清中IL-2和脑源性神经营养血液细胞因子(Brain derived neurotrophic blood cytokines,BDNF)的浓度高于联合治疗组(P<0.05)。比较治疗前后不同采血点细胞因子浓度,患者治疗2程后化疗组的IL-2浓度高于联合治疗组(P<0.05)。在不同用药周期,共有19个细胞因子出现联合治疗组高于化疗组的趋势。结论:消积饮联合FOLFOX方案可能是晚期结直肠癌的一种值得探索的治疗方案,液相芯片分析其机制可能与降低患者血清中细胞因子IL-2和BDNF水平有关。
