Objective: To explore the value of PCR-flow fluorenscence immunmicrobeads assay in prenatal gene diagnosis of thalassemia.
Methods: A total of 1001 pregnant women and their couples checked in the First Affiliated Hospital of Sun Yat-Sen University from January 2016 to August 2019 were selected. Both pregnant women and their spouses were the carriers of thalassemia gene. Samples such as amniotic fluid, were used to extract genomic DNA at the right time. Parallel detection of α- and β- thalassemia genes to samples should be carried out by PCR-flow cytometric fluorescence hybridization and traditional multiple Gap-PCR and PCR-RDB techniques. The consistency of two methods in gene diagnosis of thalassemia was evaluated by analyzing the results of detection.
Results: 389 normal genotypes (38.86%, 389/1001) and 59 abnormal genotypes (61.14%, 612/1001) was cheked out by the two methods, including 416 cases of α-thalassemia, 162 cases of β-thalassemia and 34 cases of αβ- complex thalassemia. The main genotypes of α-thalassemia were --SEA, -α3.7 and -α4.2. The mutation frequency of CD41-42 was the highest among the β-thalassemia genotypes, which followed by IVS-II-654 and CD17. A rare HKαα/--SEA thalassemia genotype was detected. Compared the traditional multiple Gap-PCR and PCR-RDB techniques, the sensitivity, specificity, positive predictive value, negative predictive value and total consistent rate of PCR-flow fluorenscence immunmicrobeads assay were 100%, which showed that the two methods were completely consistent.
Conclusion: Guangzhou is a area with high incidence of thalassemia, and the genetic types of thalassemia are complex and diverse. Prenatal diagnosis is the final barrier to the prevention of thalassemia. PCR flow-cytometric fluorescence hybridization, as a simple and fast technique, combined with traditional techniques in parallel contributed to the accuracy of prenatal gene diagnosis of thalassemia.
题目: PCR-流式荧光杂交技术在地中海贫血产前基因诊断中的应用价值.
目的: 探讨PCR-流式荧光杂交技术在地中海贫血产前基因诊断中的临床价值.
方法: 选取2016年1月-2019年8月在中山大学附属第一医院进行常规产前检查或优生遗传咨询的孕妇及其配偶1 001例，受检夫妻双方均为地中海贫血基因携带者，适时抽取羊水等样本，提取基因组DNA，采用PCR-流式荧光杂交技术和传统多重Gap-PCR和PCR-RDB技术对样本进行α和β-地中海贫血基因平行检测，分析基因诊断结果，评估两者在地中海贫血基因诊断中的一致性.
结果: 2种方法均检出正常基因型389（占38.86%，389/1001）例，异常基因型59种，共612（占61.14%，612/1001）例，包括α-地中海贫血416例、β-地中海贫血162例和αβ-复合地中海贫血34例。α-地中海贫血基因型主要为--SEA、-α3.7和-α4.2；β-地中海贫血中检出CD41-42突变频率最高，其次是IVS-II-654和CD17。检出罕见型HKαα/--SEA1例。与Gap-PCR 和PCR-RDB技术相比，PCR-流式荧光杂交法的灵敏度、特异性、阳性预测值、阴性预测值和总符合率均为100%，这2种检测方法结果一致性很好.
结论: PCR-流式荧光杂交技术操作简便快速，与传统技术联合平行检测有助于提高地中海贫血孕妇产前诊断的准确性.