Objectives:Cancer treatment relies heavily on accurate diagnosis and effective monitoring of the disease.These processes often involve invasive procedures,such as colonoscopy,to detect malignant tissues,followed by mo...Objectives:Cancer treatment relies heavily on accurate diagnosis and effective monitoring of the disease.These processes often involve invasive procedures,such as colonoscopy,to detect malignant tissues,followed by molecular analyses to determine relevant biomarkers.This study aimed to evaluate the clinical performance of droplet digital PCR(ddPCR)for detecting Kirsten Rat Sarcoma Viral Proto-Oncogene(KRAS),Neuroblastoma RAS Viral Oncogene Homolog(NRAS),and B-Raf Murine Sarcoma Viral Oncogene Homolog B(BRAF)mutations in circulating tumor DNA(ctDNA)from colorectal cancer patients using liquid biopsy.Methods:ctDNA was isolated from colorectal cancer(CRC)patients(n=110)and analyzed for KRAS,BRAF,and NRAS mutations.The ctDNA obtained through liquid biopsy was analyzed using ddPCR,and the findings were compared with sequencing data from tumor DNA archived in formalin-fixed paraffin-embedded(FFPE)blocks.Results:For KRAS mutations,ddPCR achieved a sensitivity of 72.0%and a specificity of 71.4%.However,when pooling all target mutations(KRAS,NRAS and BRAF),the overall sensitivity and specificity were lower,at 48.3%and 51.1%,respectively.Conclusion:The results of this study indicate that the ddPCR analysis of ctDNA may provide complementary information for the molecular diagnosis of CRC patients.展开更多
为了鉴定异性双胎牛性器官发育紊乱疾病——牛自由马丁(freemartinism)综合征,试验选择7头中国荷斯坦异性孪生双胎的母牛进行微滴数字PCR(droplet digital PCR,ddPCR)检测,并根据异性孪生母犊不孕的个体染色体为XX/XY嵌合、正常母牛染...为了鉴定异性双胎牛性器官发育紊乱疾病——牛自由马丁(freemartinism)综合征,试验选择7头中国荷斯坦异性孪生双胎的母牛进行微滴数字PCR(droplet digital PCR,ddPCR)检测,并根据异性孪生母犊不孕的个体染色体为XX/XY嵌合、正常母牛染色体为XX来判定是否为嵌合体。结果表明:此7个样本均存在XX/XY染色体嵌合,均为异性孪生不孕母犊。说明ddPCR方法用于鉴定牛自由马丁现象是有效的。展开更多
BACKGROUND Hypotension is a frequent complication in the intensive care unit(ICU)after adult cardiac surgery.AIM To describe frequency of hypotension in the ICU following adult cardiac surgery and its relation to the ...BACKGROUND Hypotension is a frequent complication in the intensive care unit(ICU)after adult cardiac surgery.AIM To describe frequency of hypotension in the ICU following adult cardiac surgery and its relation to the hospital outcomes.METHODS A retrospective study of post-cardiac adult surgical patients at a tertiary academic medical center in a two-year period.We abstracted baseline demographics,comorbidities,and all pertinent clinical variables.The primary predictor variable was the development of hypotension within the first 30 min upon arrival to the ICU from the operating room(OR).The primary outcome was hospital mortality,and other outcomes included duration of mechanical ventilation(MV)in hours,and ICU and hospital length of stay in days.RESULTS Of 417 patients,more than half(54%)experienced hypotension within 30 min upon arrival to the ICU.Presence of OR hypotension immediately prior to ICU transfer was significantly associated with ICU hypotension(odds ratio=1.9;95%confidence interval:1.21-2.98;P<0.006).ICU hypotensive patients had longer MV,5(interquartile ranges 3,15)vs 4 h(interquartile ranges 3,6),P=0.012.The patients who received vasopressor boluses(n=212)were more likely to experience ICU drop-off hypotension(odds ratio=1.45,95%confidence interval:0.98-2.13;P=0.062),and they experienced longer MV,ICU and hospital length of stay(P<0.001,for all).CONCLUSION Hypotension upon anesthesia-to-ICU drop-off is more frequent than previously reported and may be associated with adverse clinical outcomes.展开更多
本研究旨在建立一种检测非洲猪瘟病毒(ASFV)的微滴式数字PCR(droplet digital PCR,ddPCR)方法。通过合成ASFV特异性引物和探针,优化退火温度、引物和探针浓度等反应条件,对方法的特异性、灵敏度和重复性进行综合评价,并根据优化后的条...本研究旨在建立一种检测非洲猪瘟病毒(ASFV)的微滴式数字PCR(droplet digital PCR,ddPCR)方法。通过合成ASFV特异性引物和探针,优化退火温度、引物和探针浓度等反应条件,对方法的特异性、灵敏度和重复性进行综合评价,并根据优化后的条件进行初步临床应用。结果显示:建立的ddPCR方法最佳退火温度为61.4℃;最佳引物和探针终浓度分别为700 nmol/L和150 nmol/L;特异性试验显示该方法仅特异性扩增ASFV,与其他常见疫病病毒无交叉反应;最低检测限为1.37 copies/μL,比荧光定量PCR敏感性高10倍,浓度梯度线性分析显示良好线性关系(R^(2)=0.9898);重复性试验中变异系数均低于10%。利用该方法检测80份不同来源的疑似ASFV临床样本,结果显示均为阴性,与荧光定量聚合酶链式反应(qPCR)方法检测结果一致。综上,本研究建立的ddPCR方法敏感性高、特异性强、重复性好,可用于ASFV临床样本的精确检测分析。展开更多
基金funded by the Ministry of Health of the Czech Republic—conceptual development of research organization(MMCI,00209805)Czech Science Foundation(No.25-15990S)+1 种基金the project 7D241003 EUREKA EUROSTARS35897,project SALVAGE(P JAC,reg.No.CZ.02.01.01/00/22_008/0004644)—funded by the European Unionby the State Budget of the Czech Republic,and by the LRI project BBMRI.cz(Nos.LM2023033 and CZ.02.1.01/0.0/0.0/16_013/0001674.).
文摘Objectives:Cancer treatment relies heavily on accurate diagnosis and effective monitoring of the disease.These processes often involve invasive procedures,such as colonoscopy,to detect malignant tissues,followed by molecular analyses to determine relevant biomarkers.This study aimed to evaluate the clinical performance of droplet digital PCR(ddPCR)for detecting Kirsten Rat Sarcoma Viral Proto-Oncogene(KRAS),Neuroblastoma RAS Viral Oncogene Homolog(NRAS),and B-Raf Murine Sarcoma Viral Oncogene Homolog B(BRAF)mutations in circulating tumor DNA(ctDNA)from colorectal cancer patients using liquid biopsy.Methods:ctDNA was isolated from colorectal cancer(CRC)patients(n=110)and analyzed for KRAS,BRAF,and NRAS mutations.The ctDNA obtained through liquid biopsy was analyzed using ddPCR,and the findings were compared with sequencing data from tumor DNA archived in formalin-fixed paraffin-embedded(FFPE)blocks.Results:For KRAS mutations,ddPCR achieved a sensitivity of 72.0%and a specificity of 71.4%.However,when pooling all target mutations(KRAS,NRAS and BRAF),the overall sensitivity and specificity were lower,at 48.3%and 51.1%,respectively.Conclusion:The results of this study indicate that the ddPCR analysis of ctDNA may provide complementary information for the molecular diagnosis of CRC patients.
文摘为了鉴定异性双胎牛性器官发育紊乱疾病——牛自由马丁(freemartinism)综合征,试验选择7头中国荷斯坦异性孪生双胎的母牛进行微滴数字PCR(droplet digital PCR,ddPCR)检测,并根据异性孪生母犊不孕的个体染色体为XX/XY嵌合、正常母牛染色体为XX来判定是否为嵌合体。结果表明:此7个样本均存在XX/XY染色体嵌合,均为异性孪生不孕母犊。说明ddPCR方法用于鉴定牛自由马丁现象是有效的。
文摘BACKGROUND Hypotension is a frequent complication in the intensive care unit(ICU)after adult cardiac surgery.AIM To describe frequency of hypotension in the ICU following adult cardiac surgery and its relation to the hospital outcomes.METHODS A retrospective study of post-cardiac adult surgical patients at a tertiary academic medical center in a two-year period.We abstracted baseline demographics,comorbidities,and all pertinent clinical variables.The primary predictor variable was the development of hypotension within the first 30 min upon arrival to the ICU from the operating room(OR).The primary outcome was hospital mortality,and other outcomes included duration of mechanical ventilation(MV)in hours,and ICU and hospital length of stay in days.RESULTS Of 417 patients,more than half(54%)experienced hypotension within 30 min upon arrival to the ICU.Presence of OR hypotension immediately prior to ICU transfer was significantly associated with ICU hypotension(odds ratio=1.9;95%confidence interval:1.21-2.98;P<0.006).ICU hypotensive patients had longer MV,5(interquartile ranges 3,15)vs 4 h(interquartile ranges 3,6),P=0.012.The patients who received vasopressor boluses(n=212)were more likely to experience ICU drop-off hypotension(odds ratio=1.45,95%confidence interval:0.98-2.13;P=0.062),and they experienced longer MV,ICU and hospital length of stay(P<0.001,for all).CONCLUSION Hypotension upon anesthesia-to-ICU drop-off is more frequent than previously reported and may be associated with adverse clinical outcomes.
