目的:研究细菌培养法、免疫层析法及实时荧光定量PCR法在孕晚期B族链球菌(Group B Streptococcus,GBS)筛查中的应用价值。方法:选取2021年3月至2023年5月在我院接受产检的孕晚期孕妇154例。以病理活检为金标准,分析GBS筛查结果;对比细...目的:研究细菌培养法、免疫层析法及实时荧光定量PCR法在孕晚期B族链球菌(Group B Streptococcus,GBS)筛查中的应用价值。方法:选取2021年3月至2023年5月在我院接受产检的孕晚期孕妇154例。以病理活检为金标准,分析GBS筛查结果;对比细菌培养法、免疫层析法及实时荧光定量PCR法单独及联合诊断孕晚期GBS的效能;分析细菌培养法、免疫层析法及实时荧光定量PCR法单独及联合筛查与病理筛查结果的一致性。结果:154例孕晚期孕妇中,经病理活检检出39例孕妇GBS为阳性;三者联合诊断GBS的灵敏度、特异度、准确率、阳性预测值以及阴性预测值均高于细菌培养法、免疫层析法与实时荧光定量PCR法单独检测(P<0.05);三者联合检测与病理结果的Kappa值为0.936,均高于细菌培养法、免疫层析法、实时荧光定量PCR法单独检测。结论:在筛查孕晚期孕妇GBS中,实时荧光定量PCR法联合细菌培养法、免疫层析法诊断效能高于单独检测,可通过三者联合检测为早期诊断孕妇是否感染GBS提供依据。展开更多
目的探讨实时荧光定量聚合酶链式反应(polymerase chain reaction,PCR)技术对检测妊娠晚期B族链球菌(group B streptococcus,GBS)感染的效果。方法选取2021年1—12月江苏省丹阳市妇幼保健院接收的180例妊娠晚期孕妇,均进行实时荧光定量...目的探讨实时荧光定量聚合酶链式反应(polymerase chain reaction,PCR)技术对检测妊娠晚期B族链球菌(group B streptococcus,GBS)感染的效果。方法选取2021年1—12月江苏省丹阳市妇幼保健院接收的180例妊娠晚期孕妇,均进行实时荧光定量PCR检测,以基因测序法为金标准,分析该技术对GBS感染的诊断效能,并根据金标准将患者分为阳性组、阴性组,比较两组不良结局情况。结果实时荧光定量PCR技术检查诊断孕晚期GBS感染的准确率为99.44%、灵敏度为92.86%、特异度为100.00%,与金标准比较一致性较好(Kappa=0.96)。阳性组早产、胎膜早破、宫内感染、胎儿窘迫发生率分别为14.29%、14.29%、21.43%、21.43%,均高于阴性组,差异有统计学意义(χ^(2)=12.741、12.741、24.281、24.281,P<0.05)。阳性组新生儿感染、低体重儿发生率分别为14.29%、21.43%,均高于阴性组,差异有统计学意义(χ^(2)=12.741、24.281,P<0.05)。结论对妊娠晚期GBS感染采用实时荧光定量PCR技术检查诊断效能高,有利于预测不良妊娠结局,可指导临床诊疗工作的开展。展开更多
Late-cycle diseases (LCD) cause a significant deterioration in quality and reduce yields in soybean crops. In Argentina, in particular, leaf blight and purple seed stain, caused by the agent Cercospora kikuchii, and f...Late-cycle diseases (LCD) cause a significant deterioration in quality and reduce yields in soybean crops. In Argentina, in particular, leaf blight and purple seed stain, caused by the agent Cercospora kikuchii, and frog eye spot, caused by C. sojina, are the prevailing sources of diseases. The early, rapid and accurate detection of these phytopathogens becomes essential, and would contribute to preserving both the environment and the health of humans and animals by preventing the wasteful or improper use of chemicals such as pesticides. In order to detect Cercospora species in soybean plants at an early stage, immunochemical and molecular techniques were developed in this work. Strains from the NITE Biological Resource Center collection (Japan): Cercospora kikuchii NBRC 6711 and Cercospora sojina NBRC 6715 and regional isolates of C. kikuchii were used. To develop Dot-Blot and PCR techniques, experiments with plants undergoing different treatments were carried out: those experimentally inoculated with these fungi, those treated with sterile water and healthy plants as well. Both techniques allowed the detection, at early stages, of Cercospora species involved in two of the most frequent LCD in the country, when the cercosporin concentration produced by the fungus was higher than 3.93 ± 0.39 nmol·cyl-1 ±SD. The sensitivity between both techniques was very different. While Dot-Blot allowed the detection of the disease 4 days after inoculation, PCR detected it after 4 hours, even without visible symptoms of the disease.展开更多
The apolipoprotein E gene ε4 allele is considered a negative factor for neural regeneration in late-onset Alzheimer＇s disease cases. The aim of this study was to establish a non-invasive, rapid method to genotype ap...The apolipoprotein E gene ε4 allele is considered a negative factor for neural regeneration in late-onset Alzheimer＇s disease cases. The aim of this study was to establish a non-invasive, rapid method to genotype apolipoprotein E gene polymorphisms. Genomic DNA from mouth swab specimens was extracted using magnetic nanoparticles, and genotyping was performed by real-time PCR using TaqMan-BHQ probes. Genotyping accuracy was validated by DNA se- quencing. Our results demonstrate 100% correlation to DNA sequencing, indicating reliability of our protocol. Thus, the method we have developed for apolipoprotein E genotyping is accurate and reliable, and also suitable for genotyping large samples, which may help determine the role of the apolipoprotein E ε4 allele in neural regeneration in late-onset Alzheimer＇s disease cases.展开更多
文摘目的:研究细菌培养法、免疫层析法及实时荧光定量PCR法在孕晚期B族链球菌(Group B Streptococcus,GBS)筛查中的应用价值。方法:选取2021年3月至2023年5月在我院接受产检的孕晚期孕妇154例。以病理活检为金标准,分析GBS筛查结果;对比细菌培养法、免疫层析法及实时荧光定量PCR法单独及联合诊断孕晚期GBS的效能;分析细菌培养法、免疫层析法及实时荧光定量PCR法单独及联合筛查与病理筛查结果的一致性。结果:154例孕晚期孕妇中,经病理活检检出39例孕妇GBS为阳性;三者联合诊断GBS的灵敏度、特异度、准确率、阳性预测值以及阴性预测值均高于细菌培养法、免疫层析法与实时荧光定量PCR法单独检测(P<0.05);三者联合检测与病理结果的Kappa值为0.936,均高于细菌培养法、免疫层析法、实时荧光定量PCR法单独检测。结论:在筛查孕晚期孕妇GBS中,实时荧光定量PCR法联合细菌培养法、免疫层析法诊断效能高于单独检测,可通过三者联合检测为早期诊断孕妇是否感染GBS提供依据。
文摘目的探讨实时荧光定量聚合酶链式反应(polymerase chain reaction,PCR)技术对检测妊娠晚期B族链球菌(group B streptococcus,GBS)感染的效果。方法选取2021年1—12月江苏省丹阳市妇幼保健院接收的180例妊娠晚期孕妇,均进行实时荧光定量PCR检测,以基因测序法为金标准,分析该技术对GBS感染的诊断效能,并根据金标准将患者分为阳性组、阴性组,比较两组不良结局情况。结果实时荧光定量PCR技术检查诊断孕晚期GBS感染的准确率为99.44%、灵敏度为92.86%、特异度为100.00%,与金标准比较一致性较好(Kappa=0.96)。阳性组早产、胎膜早破、宫内感染、胎儿窘迫发生率分别为14.29%、14.29%、21.43%、21.43%,均高于阴性组,差异有统计学意义(χ^(2)=12.741、12.741、24.281、24.281,P<0.05)。阳性组新生儿感染、低体重儿发生率分别为14.29%、21.43%,均高于阴性组,差异有统计学意义(χ^(2)=12.741、24.281,P<0.05)。结论对妊娠晚期GBS感染采用实时荧光定量PCR技术检查诊断效能高,有利于预测不良妊娠结局,可指导临床诊疗工作的开展。
文摘Late-cycle diseases (LCD) cause a significant deterioration in quality and reduce yields in soybean crops. In Argentina, in particular, leaf blight and purple seed stain, caused by the agent Cercospora kikuchii, and frog eye spot, caused by C. sojina, are the prevailing sources of diseases. The early, rapid and accurate detection of these phytopathogens becomes essential, and would contribute to preserving both the environment and the health of humans and animals by preventing the wasteful or improper use of chemicals such as pesticides. In order to detect Cercospora species in soybean plants at an early stage, immunochemical and molecular techniques were developed in this work. Strains from the NITE Biological Resource Center collection (Japan): Cercospora kikuchii NBRC 6711 and Cercospora sojina NBRC 6715 and regional isolates of C. kikuchii were used. To develop Dot-Blot and PCR techniques, experiments with plants undergoing different treatments were carried out: those experimentally inoculated with these fungi, those treated with sterile water and healthy plants as well. Both techniques allowed the detection, at early stages, of Cercospora species involved in two of the most frequent LCD in the country, when the cercosporin concentration produced by the fungus was higher than 3.93 ± 0.39 nmol·cyl-1 ±SD. The sensitivity between both techniques was very different. While Dot-Blot allowed the detection of the disease 4 days after inoculation, PCR detected it after 4 hours, even without visible symptoms of the disease.
基金supported by two grants from Science,Industry,Trade and Information Technology Commission of Shenzhen Municipality in China,grant No.201002063,JC20110518075 7A
文摘The apolipoprotein E gene ε4 allele is considered a negative factor for neural regeneration in late-onset Alzheimer＇s disease cases. The aim of this study was to establish a non-invasive, rapid method to genotype apolipoprotein E gene polymorphisms. Genomic DNA from mouth swab specimens was extracted using magnetic nanoparticles, and genotyping was performed by real-time PCR using TaqMan-BHQ probes. Genotyping accuracy was validated by DNA se- quencing. Our results demonstrate 100% correlation to DNA sequencing, indicating reliability of our protocol. Thus, the method we have developed for apolipoprotein E genotyping is accurate and reliable, and also suitable for genotyping large samples, which may help determine the role of the apolipoprotein E ε4 allele in neural regeneration in late-onset Alzheimer＇s disease cases.
