The Application Of Triple TaqMan Real-time Fluorescence Quantitative PCR In The Clinical Diagnosis Of Deletion ?-thalassemia

Objective:The clinical samples were detected by triple TaqMan real-time fluorescence quantitative PCR system,which was successfully established by our research team,to detect the clinical samples,and to explore the positive detection of common and uncommon deletion?-thalassemia in premarital examination,pregnancy examination,eugenics genetic clinic,etc of positive detection rate and genotype distribution.Methods:From August 2018 to July 2019,people who went to the Affiliated Hospital of Youjiang Medical College for Nationalities and Baise maternal and child health care hospital of Baise City for premarital examination,pregnancy examination,eugenic genetic counseling,and thalassemia gene detection were selected as the research objects,and peripheral blood samples were collected.In addition,the clinical data of mean corpuscular volume?MCV?,mean corpuscular hemoglobin?MCH?and hemoglobin A₂?HbA₂?were collected.The blood routine test and hemoglobin electrophoresis examination were improved for some blank data.The deletion?-thalassemia gene was detected by triple TaqMan real-time quantitative PCR and gap PCR.The results of MCV,MCH and HbA₂ were analyzed and compared.Results:?1?A total of 1568 samples of thalassemia gene detection were included in this study.452 cases of deletion type?-thalassemia were detected by triple TaqMan real-time fluorescence quantitative PCR and the detection rate was 28.83%.Among them,there were 164 cases silent type,250 cases light type and 38 cases middle type.?2?The results of gap PCR analysis showed that there were 126 cases with-?^3.7/??and 38 cases with-?^4.2/??in 164 silent type,243 cases with--^SEA/???5 cases with-?^3.7/-?^3.7?1 case with-?^3.7/-?^4.2 and 1 case with--^Thai/??in 250 light type and 28 cases with--^SEA/-?^3.7?8 cases with--^SEA/-?^4.2?1 case with--^Thai/-?^3.7 and1 case with--^SEA/-?^21.9in 38 cases middle type.?3?The missed diagnosis rate of MCV,MCH and HbA₂ was higher than 10%,and the missed diagnosis rate of quiescent?-thalassemia was the highest,in particular.Conclusion:?1?This study suggests that triple TaqMan real-time fluorescence quantitative PCR can detect not only common deletion type of?-thalassemia,but also the rare deletion type?-thalassemia.?2?Compared with traditional screening methods,triple TaqMan real-time fluorescence quantitative PCR can reduce missed diagnosis,especially for quiescent?-thalassemia.