A set of allele-specific oligonucleotide (ASO) probes used for detecting all 18 β-tha-lassemia mutations found in Chinese was immobilized on two strips of Biodyne C membrane;one containing 7 pairs of oligonucleotide ...A set of allele-specific oligonucleotide (ASO) probes used for detecting all 18 β-tha-lassemia mutations found in Chinese was immobilized on two strips of Biodyne C membrane;one containing 7 pairs of oligonucleotide probes specific for the most commonly found mutant al-leles,and the other containing the remaining 11 pairs of ASO_s specific for the less commonlyfound.The membranes were hybridized with β-globin sequences amplified by polymerase chainreaction (PCR) with biotinylated primers,and then treated with Streptavidin-POD conjugateand substrates for color development.The method has been applied successfully to the detectionof all 18 Chinese β-thalassemia mutations and prenatal diagnosis of two high-risk pregnancies ofβ-thalassemia.Patients with homozygous,heterozygous and compound heterozygous alleles ofthese mutations and normal individuals could be easily distinguished by the present method.Us-ing the immobilized-probe format (reverse dot blot),it was able to screen simultaneously multi-ple β-thalassemia mutations of a DNA sample by performing hybridization only once.This assayis simple,rapid and independent of radio-isotopes and can be appplied for all 18 β-thalassemiamutations so far found in Chinese population.It is considered that this method may be usefulfor gene frequency investigation of large numbers of β-thalassemia DNA samples and used as aroutine method in the clinic laboratory.展开更多
A technique of direct sequence analysis of β-globin gene with the products of amplifi-cation by polymerase chain reaction (PCR) was reported and a case of β-thalassemia with therare mutation in Chinese,‘codon 14/15...A technique of direct sequence analysis of β-globin gene with the products of amplifi-cation by polymerase chain reaction (PCR) was reported and a case of β-thalassemia with therare mutation in Chinese,‘codon 14/15 (+G)’ was detected by this method.After the se-quence of the mutation site was determined,an analysis of the restriction map of the gene anddot blot hybridization with radioactive allele specific oligonucleotide probe was designed to con-firm the result of DNA sequencing.展开更多
目的：探讨胎儿脐血血红蛋白电泳Hb A值在孕中、晚期诊断β-地中海贫血以及母血污染鉴定的应用价值。方法收集父母双方为β-地中海贫血携带者作为指征进行产前诊断且孕周为24～28 w的胎儿54例。应用全自动毛细管电泳系统对胎儿脐带血进...目的：探讨胎儿脐血血红蛋白电泳Hb A值在孕中、晚期诊断β-地中海贫血以及母血污染鉴定的应用价值。方法收集父母双方为β-地中海贫血携带者作为指征进行产前诊断且孕周为24～28 w的胎儿54例。应用全自动毛细管电泳系统对胎儿脐带血进行电泳分析，且采用反向点杂交法检测β地中海贫血基因。结果12例β-地中海贫血基因型正常的胎儿脐带血 Hb A 的平均值为（6.3％±1.9％）；31例β杂合突变的 Hb A 值范围为2.6％～3.7％；10例基因纯合突变或双重杂合突变中有6例胎血 Hb A 为0，4例为0.4％～0.7％；母血污染1例。结论脐血血红蛋白电泳Hb A值可辅助诊断轻型、中间型及重型胎儿β-地中海贫血，并可以作为母血污染鉴定的一种方法。展开更多
目的评估红细胞指数对鉴别婚育年龄轻型β地中海贫血和缺铁性贫血患者的作用，并利用这些指数建立基于反向传播（BP）人工神经网络的诊断模型以提高预测准确率。方法选择310例轻型β地中海贫血患者，其中男性131例．女性179例．年龄23...目的评估红细胞指数对鉴别婚育年龄轻型β地中海贫血和缺铁性贫血患者的作用，并利用这些指数建立基于反向传播（BP）人工神经网络的诊断模型以提高预测准确率。方法选择310例轻型β地中海贫血患者，其中男性131例．女性179例．年龄23～37岁，中位年龄25岁：125例缺铁性贫血患者作为对照，其中男性52例，女性73例，年龄20～34岁，中位年龄24岁。采用7种公式计算红细胞指数，分析它们对轻型β地中海贫血的预测灵敏度、特异度和约登指数，以此7种公式的计算值作为参数在MATLAB7．0程序上实现BP人工神经网络诊断模型．结果轻型β地贫血红细胞和血红蛋白（Hb）水平比缺铁性贫血高，平均红细胞体积（MCV）、平均红细胞血红蛋白（MCH）比后者低。7种指数中．Green and King指数（GKI）对轻型β地中海贫血的预测灵敏度（90．6％）及约登指数（78．6％）最高。BP人工神经网络诊断模型刈轻型β地中海贫血的诊断准确率为96．77％。结论GKI用于鉴别轻型β地中海贫血和缺铁性贫血可靠性相对较高。展开更多
文摘A set of allele-specific oligonucleotide (ASO) probes used for detecting all 18 β-tha-lassemia mutations found in Chinese was immobilized on two strips of Biodyne C membrane;one containing 7 pairs of oligonucleotide probes specific for the most commonly found mutant al-leles,and the other containing the remaining 11 pairs of ASO_s specific for the less commonlyfound.The membranes were hybridized with β-globin sequences amplified by polymerase chainreaction (PCR) with biotinylated primers,and then treated with Streptavidin-POD conjugateand substrates for color development.The method has been applied successfully to the detectionof all 18 Chinese β-thalassemia mutations and prenatal diagnosis of two high-risk pregnancies ofβ-thalassemia.Patients with homozygous,heterozygous and compound heterozygous alleles ofthese mutations and normal individuals could be easily distinguished by the present method.Us-ing the immobilized-probe format (reverse dot blot),it was able to screen simultaneously multi-ple β-thalassemia mutations of a DNA sample by performing hybridization only once.This assayis simple,rapid and independent of radio-isotopes and can be appplied for all 18 β-thalassemiamutations so far found in Chinese population.It is considered that this method may be usefulfor gene frequency investigation of large numbers of β-thalassemia DNA samples and used as aroutine method in the clinic laboratory.
文摘A technique of direct sequence analysis of β-globin gene with the products of amplifi-cation by polymerase chain reaction (PCR) was reported and a case of β-thalassemia with therare mutation in Chinese,‘codon 14/15 (+G)’ was detected by this method.After the se-quence of the mutation site was determined,an analysis of the restriction map of the gene anddot blot hybridization with radioactive allele specific oligonucleotide probe was designed to con-firm the result of DNA sequencing.
文摘目的：探讨胎儿脐血血红蛋白电泳Hb A值在孕中、晚期诊断β-地中海贫血以及母血污染鉴定的应用价值。方法收集父母双方为β-地中海贫血携带者作为指征进行产前诊断且孕周为24～28 w的胎儿54例。应用全自动毛细管电泳系统对胎儿脐带血进行电泳分析，且采用反向点杂交法检测β地中海贫血基因。结果12例β-地中海贫血基因型正常的胎儿脐带血 Hb A 的平均值为（6.3％±1.9％）；31例β杂合突变的 Hb A 值范围为2.6％～3.7％；10例基因纯合突变或双重杂合突变中有6例胎血 Hb A 为0，4例为0.4％～0.7％；母血污染1例。结论脐血血红蛋白电泳Hb A值可辅助诊断轻型、中间型及重型胎儿β-地中海贫血，并可以作为母血污染鉴定的一种方法。
文摘目的评估红细胞指数对鉴别婚育年龄轻型β地中海贫血和缺铁性贫血患者的作用，并利用这些指数建立基于反向传播（BP）人工神经网络的诊断模型以提高预测准确率。方法选择310例轻型β地中海贫血患者，其中男性131例．女性179例．年龄23～37岁，中位年龄25岁：125例缺铁性贫血患者作为对照，其中男性52例，女性73例，年龄20～34岁，中位年龄24岁。采用7种公式计算红细胞指数，分析它们对轻型β地中海贫血的预测灵敏度、特异度和约登指数，以此7种公式的计算值作为参数在MATLAB7．0程序上实现BP人工神经网络诊断模型．结果轻型β地贫血红细胞和血红蛋白（Hb）水平比缺铁性贫血高，平均红细胞体积（MCV）、平均红细胞血红蛋白（MCH）比后者低。7种指数中．Green and King指数（GKI）对轻型β地中海贫血的预测灵敏度（90．6％）及约登指数（78．6％）最高。BP人工神经网络诊断模型刈轻型β地中海贫血的诊断准确率为96．77％。结论GKI用于鉴别轻型β地中海贫血和缺铁性贫血可靠性相对较高。
