Bones and teeth often represent the only sources of DNA available for identifying human remains.DNA in bones and teeth is generally better preserved than that in soft tissues because of the presence of hard connective...Bones and teeth often represent the only sources of DNA available for identifying human remains.DNA in bones and teeth is generally better preserved than that in soft tissues because of the presence of hard connective tissue with a high level of calcium.Because of the extensive mineralisation,the choice of an efficient DNA extraction procedure is important to minimise the sampling of a high level of minerals and to remove polymerase chain reaction(PCR)inhibitors.Some protocols are available for DNA extraction from bones and teeth as part of the Qiagen EZ1 DNA Investigator Kit using the EZ1 Advanced XL automated purification platform.To improve the efficiency of DNA extraction from skeletal remains,the present study focuses on a modification to these already available protocols.In this study,different bones and teeth collected between 1 and 50 years after death were subjected to DNA extraction using the standard EZ1 protocol,a supplementary protocol,and a modified protocol.The modified approach included a decalcification step,whereas the Qiagen protocols worked directly on non-decalcified powder.In all three procedures,150 mg samples were used for DNA extraction.We evaluated the quantity of DNA recovered from samples,the presence of any PCR inhibitors co-extracted,the level of DNA degradation,the quality of short tandem repeat(STR)profiles,and the reproducibility of the modified procedure.When compared with the other protocols,the modified protocol resulted in the best recovery of DNA that was free of PCR inhibitors.Additionally,the STR profiles were reliable and of high quality.In our opinion,the decalcification step increases DNA recovery by softening tissues,which allows lysis solutions to act more effectively.Furthermore,the use of two lysis solutions and the variation added to the EZ1 purification step allow for DNA recovery with quality and quantity superior to those of the previously available Qiagen-based protocols.These findings may be helpful solutions to the problems commonly encountered when dealing with difficult samples,such as bones and teeth.展开更多
目的:对博唐平®A1C EZ 2.0糖化血红蛋白(HbA1c)床旁检测仪的临床性能进行评估。方法:选取2020年4-8月广东省人民医院收治的100例糖尿病患者为研究对象,将实验室仪器设为参考系统,博唐平®A1C EZ 2.0设为评估系统,对两款仪器检...目的:对博唐平®A1C EZ 2.0糖化血红蛋白(HbA1c)床旁检测仪的临床性能进行评估。方法:选取2020年4-8月广东省人民医院收治的100例糖尿病患者为研究对象,将实验室仪器设为参考系统,博唐平®A1C EZ 2.0设为评估系统,对两款仪器检测HbA1c的结果进行Passing-Bablok回归分析、Bland-Altman分析、偏差分析。结果:博唐平®A1C EZ 2.0指尖末梢血组与博唐平®A1C EZ 2.0静脉血组比较,再与实验室静脉血组比较,结果均呈线性相关(r=0.946);博唐平®A1C EZ 2.0所测结果均值与实验室Bio-RadD-10 HbA1c测定仪所测结果均值比较,平均相对偏差为0.2%,95%CI内相对偏差为-1.7%~2.1%,仅有6.00%(6/100)界外点数。结论:博唐平®A1C EZ 2.0能够满足HbA1c临床检测的应用要求,适用于临床糖尿病管理。展开更多
文摘Bones and teeth often represent the only sources of DNA available for identifying human remains.DNA in bones and teeth is generally better preserved than that in soft tissues because of the presence of hard connective tissue with a high level of calcium.Because of the extensive mineralisation,the choice of an efficient DNA extraction procedure is important to minimise the sampling of a high level of minerals and to remove polymerase chain reaction(PCR)inhibitors.Some protocols are available for DNA extraction from bones and teeth as part of the Qiagen EZ1 DNA Investigator Kit using the EZ1 Advanced XL automated purification platform.To improve the efficiency of DNA extraction from skeletal remains,the present study focuses on a modification to these already available protocols.In this study,different bones and teeth collected between 1 and 50 years after death were subjected to DNA extraction using the standard EZ1 protocol,a supplementary protocol,and a modified protocol.The modified approach included a decalcification step,whereas the Qiagen protocols worked directly on non-decalcified powder.In all three procedures,150 mg samples were used for DNA extraction.We evaluated the quantity of DNA recovered from samples,the presence of any PCR inhibitors co-extracted,the level of DNA degradation,the quality of short tandem repeat(STR)profiles,and the reproducibility of the modified procedure.When compared with the other protocols,the modified protocol resulted in the best recovery of DNA that was free of PCR inhibitors.Additionally,the STR profiles were reliable and of high quality.In our opinion,the decalcification step increases DNA recovery by softening tissues,which allows lysis solutions to act more effectively.Furthermore,the use of two lysis solutions and the variation added to the EZ1 purification step allow for DNA recovery with quality and quantity superior to those of the previously available Qiagen-based protocols.These findings may be helpful solutions to the problems commonly encountered when dealing with difficult samples,such as bones and teeth.
文摘目的:对博唐平®A1C EZ 2.0糖化血红蛋白(HbA1c)床旁检测仪的临床性能进行评估。方法:选取2020年4-8月广东省人民医院收治的100例糖尿病患者为研究对象,将实验室仪器设为参考系统,博唐平®A1C EZ 2.0设为评估系统,对两款仪器检测HbA1c的结果进行Passing-Bablok回归分析、Bland-Altman分析、偏差分析。结果:博唐平®A1C EZ 2.0指尖末梢血组与博唐平®A1C EZ 2.0静脉血组比较,再与实验室静脉血组比较,结果均呈线性相关(r=0.946);博唐平®A1C EZ 2.0所测结果均值与实验室Bio-RadD-10 HbA1c测定仪所测结果均值比较,平均相对偏差为0.2%,95%CI内相对偏差为-1.7%~2.1%,仅有6.00%(6/100)界外点数。结论:博唐平®A1C EZ 2.0能够满足HbA1c临床检测的应用要求,适用于临床糖尿病管理。
