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
Pseudomonas aeruginosa(P:aeruginosa)is a major opportunistic pathogen in hospital-acquired infections.Thus,carly diagnosis is the best strategy for fighting against these infections.In this report,we incorporated mult...Pseudomonas aeruginosa(P:aeruginosa)is a major opportunistic pathogen in hospital-acquired infections.Thus,carly diagnosis is the best strategy for fighting against these infections.In this report,we incorporated multiple crOsS displacement amplification(MCDA)combined with the malachite green(MG)for rapid,sensitive,specific and visual detection of P.aeruginosa by targeting the oprl gene.The MCDA-MG assay was conducted at 67°C for only 40 min during the amplification stage,and then products were directly detected by using colorimetric indicators(MG),eliminating the use of an electrophoresis instrument or amplicon analysis equipment.The entire process,including specimen processing(35 min),amplification(40 min)and detection(5 min),can be finished within 80 min.All 30 non-P.aeruginosa strains tested negative,indicating the high specificity of the MCDA primers.The analytical sensitivity of the MCDA-MG assay was 100 fg of genomic templates per reaction in pure culture,which was in complete accordance with MCDA by gel clectrophoresis and real-time turbidity.The assay was also successfully applied to detecting P.aeruginosa in stool samples.Therefore,the rapidity,simplicity,and nearly equipment free platform of the MCDA-MG technique make it possible for clinical diagnosis,and more.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Pseudomonas aeruginosa(P:aeruginosa)is a major opportunistic pathogen in hospital-acquired infections.Thus,carly diagnosis is the best strategy for fighting against these infections.In this report,we incorporated multiple crOsS displacement amplification(MCDA)combined with the malachite green(MG)for rapid,sensitive,specific and visual detection of P.aeruginosa by targeting the oprl gene.The MCDA-MG assay was conducted at 67°C for only 40 min during the amplification stage,and then products were directly detected by using colorimetric indicators(MG),eliminating the use of an electrophoresis instrument or amplicon analysis equipment.The entire process,including specimen processing(35 min),amplification(40 min)and detection(5 min),can be finished within 80 min.All 30 non-P.aeruginosa strains tested negative,indicating the high specificity of the MCDA primers.The analytical sensitivity of the MCDA-MG assay was 100 fg of genomic templates per reaction in pure culture,which was in complete accordance with MCDA by gel clectrophoresis and real-time turbidity.The assay was also successfully applied to detecting P.aeruginosa in stool samples.Therefore,the rapidity,simplicity,and nearly equipment free platform of the MCDA-MG technique make it possible for clinical diagnosis,and more.
