Harmful cyanobacterial blooms, especially Microcystis blooms, occur worldwide and draw widespread attention. The dynamics of microcystin-producing Microcystis and competition between microcystin-producing Microcystis ...Harmful cyanobacterial blooms, especially Microcystis blooms, occur worldwide and draw widespread attention. The dynamics of microcystin-producing Microcystis and competition between microcystin-producing Microcystis and non-microcystin-producing Microcystis are key to predicting and treating Microcystis blooms. Multiplex qPCR is a useful tool to assess such issues. In this study, we developed multiplex qPCR methods with newly-designed probes and primers for the microcystin-synthesis related genes mcyA and mcyE. We used seven toxic Microcystis strains and four non-toxic Microcystis strains to compare the differences in the ratios of toxic and non-toxic Microcystis in mixed cultures, which were calculated using abundances of the genes mcyA, mcyB, mcyD, mcy E and phycocyanin( PC). We also compared traditional cell counting and multiplex qPCR. Hierarchical clustering and principal component analysis indicated that mcyD was the most suitable mcy gene for quantification in laboratory experiments. mcyB abundances were always higher; we suggest that the amount of toxic Microcystis measured using mcyB might overestimate the actual percentages.展开更多
Human herpesviruses are double-stranded DNA viruses that are classified into nine species.More than 90%of adults are ever infected with one or more herpesviruses.The symptoms of infection with different herpesviruses ...Human herpesviruses are double-stranded DNA viruses that are classified into nine species.More than 90%of adults are ever infected with one or more herpesviruses.The symptoms of infection with different herpesviruses are diverse ranging from mild or asymptomatic infections to deadly diseases such as aggressive lymphomas and sarcomas.Timely and accurate detection of herpesvirus infection is critical for clinical management and treatment.In this study,we established a single-tube nonuple qPCR assay for detection of all nine herpesviruses using a 2-D multiplex qPCR method with a house-keeping gene as the internal control.The novel assay can detect and distinguish different herpesviruses with 30 to 300 copies per 25µL single-tube reaction,and does not cross-react with 20 other human viruses,including DNA and RNA viruses.The robustness of the novel assay was evaluated using 170 clinical samples.The novel assay showed a high consistency(100%)with the single qPCR assay for HHVs detection.The features of simple,rapid,high sensitivity,specificity,and low cost make this assay a high potential to be widely used in clinical diagnosis and patient treatment.展开更多
目的运用新西兰政府初级产业部(Ministry of Primary Industry, MPI)提出的多重qPCR方法鉴定麦卢卡蜂蜜。方法参照MPI提出的多重qPCR方法,建立麦卢卡标准曲线,提取15个未知来源的蜂蜜样品样品中的花粉DNA,进行样品多重qPCR检测分析。结...目的运用新西兰政府初级产业部(Ministry of Primary Industry, MPI)提出的多重qPCR方法鉴定麦卢卡蜂蜜。方法参照MPI提出的多重qPCR方法,建立麦卢卡标准曲线,提取15个未知来源的蜂蜜样品样品中的花粉DNA,进行样品多重qPCR检测分析。结果建立麦卢卡标准曲线获得Manuka通道的阈值范围,可以用于今后蜂蜜样品的麦卢卡特异性分析。M2-4蜂蜜样品中的花粉DNA不来自麦卢卡树Leptospermum scoparium。结论本研究对MPI提出的鉴定麦卢卡蜂蜜方法起到很好的推动及指导作用。展开更多
基金Supported by the National Natural Science Foundation of China(Nos.31370418,41561144008)the Jiangxi Water Science and Technology Fund(No.KT201602)the State Key Laboratory of Freshwater Ecology and Biotechnology(No.2016FBZ07)
文摘Harmful cyanobacterial blooms, especially Microcystis blooms, occur worldwide and draw widespread attention. The dynamics of microcystin-producing Microcystis and competition between microcystin-producing Microcystis and non-microcystin-producing Microcystis are key to predicting and treating Microcystis blooms. Multiplex qPCR is a useful tool to assess such issues. In this study, we developed multiplex qPCR methods with newly-designed probes and primers for the microcystin-synthesis related genes mcyA and mcyE. We used seven toxic Microcystis strains and four non-toxic Microcystis strains to compare the differences in the ratios of toxic and non-toxic Microcystis in mixed cultures, which were calculated using abundances of the genes mcyA, mcyB, mcyD, mcy E and phycocyanin( PC). We also compared traditional cell counting and multiplex qPCR. Hierarchical clustering and principal component analysis indicated that mcyD was the most suitable mcy gene for quantification in laboratory experiments. mcyB abundances were always higher; we suggest that the amount of toxic Microcystis measured using mcyB might overestimate the actual percentages.
基金supported by the grants from the National Science and Technology Major Project of China (2019YFC1200603, and 2017ZX10103009-002)
文摘Human herpesviruses are double-stranded DNA viruses that are classified into nine species.More than 90%of adults are ever infected with one or more herpesviruses.The symptoms of infection with different herpesviruses are diverse ranging from mild or asymptomatic infections to deadly diseases such as aggressive lymphomas and sarcomas.Timely and accurate detection of herpesvirus infection is critical for clinical management and treatment.In this study,we established a single-tube nonuple qPCR assay for detection of all nine herpesviruses using a 2-D multiplex qPCR method with a house-keeping gene as the internal control.The novel assay can detect and distinguish different herpesviruses with 30 to 300 copies per 25µL single-tube reaction,and does not cross-react with 20 other human viruses,including DNA and RNA viruses.The robustness of the novel assay was evaluated using 170 clinical samples.The novel assay showed a high consistency(100%)with the single qPCR assay for HHVs detection.The features of simple,rapid,high sensitivity,specificity,and low cost make this assay a high potential to be widely used in clinical diagnosis and patient treatment.
