A set of universal loop-mediated isothermal amplification （LAMP） primers targeting the flo gene was designed to detect Borrelia burgdorferi sensu lato （B. burgdorferi s.I.） in human samples. The sensitivity of LAM...A set of universal loop-mediated isothermal amplification （LAMP） primers targeting the flo gene was designed to detect Borrelia burgdorferi sensu lato （B. burgdorferi s.I.） in human samples. The sensitivity of LAMP was 20 copies/reaction, and the assay did not detect false positives among 11 other related bacteria. A positive LAMP result was obtained for 9 of the 24 confirmed cases and for 12 of 94 suspected cases. The positive rate of LAMP was the same as that of nested PCR. The LAMP is a useful diagnostic method that can be developed for rapid detection of B. burgdorferi s.I. in human sera. Combination of the LAMP and nested PCR was more sensitive for detecting B. burgdorferi s.I. in human serum samples.展开更多
目的:探讨体外CD10^(+)干细胞与VSV病毒共培养分化后,能否从分化后细胞的基因组中扩增出成熟抗体IgG重链(VH)、轻链(VL)可变区基因片段。方法:37℃培养CD10^(+)干细胞至单层细胞,B细胞扩增培养基及添加的白介素因子、VSV病毒与CD10^(+)...目的:探讨体外CD10^(+)干细胞与VSV病毒共培养分化后,能否从分化后细胞的基因组中扩增出成熟抗体IgG重链(VH)、轻链(VL)可变区基因片段。方法:37℃培养CD10^(+)干细胞至单层细胞,B细胞扩增培养基及添加的白介素因子、VSV病毒与CD10^(+)干细胞共培养15 d,留存每天细胞上清测定IgM含量,以15 d细胞为样本抽提总RNA,反转录为cDNA,以其为模板,用单链抗体(ScFv)噬菌体抗体库构建引物扩增VH、VL。结果:利用含VSV病毒、IL-2、IL-4、IL-7、IL-10、IL-21、CD40L的B细胞扩增培养基成功将CD10^(+)干细胞分化为CD38、CD138阳性浆细胞,IgM含量随时间递减,诱导15 d后成功扩增出成熟抗体IgG的370 bp VH、320 bp VL可变区基因片段。结论:成功完成了基于CD10^(+)干细胞的人源抗体基因获取,可用于各类人畜共患病病毒噬菌体抗体库构建。展开更多
Background Malaria continues to cause burden in various parts of the world.Haiti,a Caribbean country,is among those aiming to eliminate malaria within a few years.Two surveys were conducted in Haiti during which we ai...Background Malaria continues to cause burden in various parts of the world.Haiti,a Caribbean country,is among those aiming to eliminate malaria within a few years.Two surveys were conducted in Haiti during which we aimed to evaluate the performance of the simple and rapid procedure for ultra-rapid extraction-loop-mediated isothermal amplifcation(PURE-LAMP)method with dried blood spots as an alternative diagnostic method for malaria in the context of low to very low rates of transmission.Methods Febrile and afebrile people were recruited from three administrative divisions within Haiti:Nippes,Sud and Grand’Anse,during the summers of 2017(early August to early September)and 2018(late July to late August).Their blood samples were tested by microscopy,rapid diagnostic tests(RDT),PURE-LAMP and nested PCR to detect Plasmodium infection.Sensitivity,specifcity,positive and negative predictive values and kappa statistics were estimated with the nested PCR results as the gold standard.Results Among 1074 samples analyzed,a positive rate of 8.3%was calculated based on the nested PCR results.Among febrile participants,the rates in 2017 and 2018 were 14.6%and 1.4%,respectively.Three positives were detected among 172 afebrile participants in 2018 by PURE-LAMP and nested PCR,and all three were from the same locality.There was no afebrile participants recruited in 2017.The PURE-LAMP,RDT and microscopy had respective sensitivities of 100%,85.4%and 49.4%.All of the testing methods had specifcities over 99%.Conclusions This study confrmed the high performance of the PURE-LAMP method to detect Plasmodium infection with dried blood spots and recommends its use in targeted mass screening and treatment activities in low endemic areas of malaria.展开更多
基金funded by the National Key Science and Technology Projects of China(2012ZX10004219 and 2013ZX10004001)
文摘A set of universal loop-mediated isothermal amplification （LAMP） primers targeting the flo gene was designed to detect Borrelia burgdorferi sensu lato （B. burgdorferi s.I.） in human samples. The sensitivity of LAMP was 20 copies/reaction, and the assay did not detect false positives among 11 other related bacteria. A positive LAMP result was obtained for 9 of the 24 confirmed cases and for 12 of 94 suspected cases. The positive rate of LAMP was the same as that of nested PCR. The LAMP is a useful diagnostic method that can be developed for rapid detection of B. burgdorferi s.I. in human sera. Combination of the LAMP and nested PCR was more sensitive for detecting B. burgdorferi s.I. in human serum samples.
文摘目的:探讨体外CD10^(+)干细胞与VSV病毒共培养分化后,能否从分化后细胞的基因组中扩增出成熟抗体IgG重链(VH)、轻链(VL)可变区基因片段。方法:37℃培养CD10^(+)干细胞至单层细胞,B细胞扩增培养基及添加的白介素因子、VSV病毒与CD10^(+)干细胞共培养15 d,留存每天细胞上清测定IgM含量,以15 d细胞为样本抽提总RNA,反转录为cDNA,以其为模板,用单链抗体(ScFv)噬菌体抗体库构建引物扩增VH、VL。结果:利用含VSV病毒、IL-2、IL-4、IL-7、IL-10、IL-21、CD40L的B细胞扩增培养基成功将CD10^(+)干细胞分化为CD38、CD138阳性浆细胞,IgM含量随时间递减,诱导15 d后成功扩增出成熟抗体IgG的370 bp VH、320 bp VL可变区基因片段。结论:成功完成了基于CD10^(+)干细胞的人源抗体基因获取,可用于各类人畜共患病病毒噬菌体抗体库构建。
文摘Background Malaria continues to cause burden in various parts of the world.Haiti,a Caribbean country,is among those aiming to eliminate malaria within a few years.Two surveys were conducted in Haiti during which we aimed to evaluate the performance of the simple and rapid procedure for ultra-rapid extraction-loop-mediated isothermal amplifcation(PURE-LAMP)method with dried blood spots as an alternative diagnostic method for malaria in the context of low to very low rates of transmission.Methods Febrile and afebrile people were recruited from three administrative divisions within Haiti:Nippes,Sud and Grand’Anse,during the summers of 2017(early August to early September)and 2018(late July to late August).Their blood samples were tested by microscopy,rapid diagnostic tests(RDT),PURE-LAMP and nested PCR to detect Plasmodium infection.Sensitivity,specifcity,positive and negative predictive values and kappa statistics were estimated with the nested PCR results as the gold standard.Results Among 1074 samples analyzed,a positive rate of 8.3%was calculated based on the nested PCR results.Among febrile participants,the rates in 2017 and 2018 were 14.6%and 1.4%,respectively.Three positives were detected among 172 afebrile participants in 2018 by PURE-LAMP and nested PCR,and all three were from the same locality.There was no afebrile participants recruited in 2017.The PURE-LAMP,RDT and microscopy had respective sensitivities of 100%,85.4%and 49.4%.All of the testing methods had specifcities over 99%.Conclusions This study confrmed the high performance of the PURE-LAMP method to detect Plasmodium infection with dried blood spots and recommends its use in targeted mass screening and treatment activities in low endemic areas of malaria.
