Diagnosis of the accurate genotype of HKαα carriers in patients with thalassemia using multiplex ligation-dependent probe amplification combined with nested polymerase chain reaction

作     者：Dong-Mei Chen Shi Ma Xiang-Lan Tang Ji-Yun Yang Zheng-Lin Yang Chen Dong-Mei;Ma Shi;Tang Xiang-Lan;Yang Ji-Yun;Yang Zheng-Lin

作者机构：Clinical Medical SchoolSouthwest Medical UniversityLuzhouSichuan 646000China The Key Laboratory for Human Disease Gene Study of Sichuan Province and Institute of Laboratory MedicineSichuan Provincial People's HospitalUniversity of Electronic Science and Technology of ChinaChengduSichuan 610072China Research Unit for Blindness Prevention of Chinese Academy of Medical Sciences(2019RU026)Sichuan Academy of Medical SciencesChengduSichuan 610072China Natural Products Research CenterInstitute of Chengdu BiologySichuan Translational Medicine HospitalChinese Academy of SciencesChengduSichuan 610072China. 

出 版 物：《Chinese Medical Journal》 (中华医学杂志（英文版）)

年 卷 期：2020年第133卷第10期

页      面：1175-1181页

核心收录：

学科分类：1002[医学-临床医学] 100201[医学-内科学(含：心血管病、血液病、呼吸系病、消化系病、内分泌与代谢病、肾病、风湿病、传染病)] 10[医学] 

基　　金：This work was supported by a grant from the Department of Science and Technology of Sichuan province China(No.30504010332) 

主　　题：Thalassemia HongKongαα Nested polymerase chain reaction Multiplex ligation-dependent probe amplification Gene dosage 

摘      要：Background:Patients carrying the HongKongαα(HKαα)allele and-α3.7/αααanti-4.2 could be misdiagnosed as-α3.7/ααby the current conventional thalassemia detection methods,leading to inaccurate genetic counseling and an incorrect prenatal *** study was aimed to accurately analyze the genotypes of HKααcarriers and-α3.7/ααα***::Samples were collected in our hospital from July 2017 to October ***-four common types of Chinese thalassemia were screened by gap-polymerase chain reaction(Gap-PCR)and reverse dot blot(RDB).Anti-4.2 multiplex-PCR was used to confirm carriers of theαααanti-4.2 duplication with-α3.7 ***-round nested PCR and multiplex ligation-dependent probe amplification(MLPA)were applied to accurately identify and confirm their *** data analysis,we used descriptive statistics and Fisher’s exact ***::Two thousand five hundred and forty-four cases were identified as thalassemia in 5488 peripheral blood *** results showed thatα,β,andαβcompound thalassemia were identified in 1190(46.78%),1286(50.55%),and 68(2.67%)cases,respectively.A total of 227 samples from thalassemia patients were identified as-α3.7/ααby Gap-PCR,and the genotypes of two samples were *** was a difference between Gap-PCR and combined groups(Gap-PCR combined with nested PCR and MLPA)in detecting HKαα(P0.05).Among the 229 patients,20 patients were identified as HKααcarriers and one was identified as-α3.7/ααα anti-4.2 by two-round nested PCR and MLPA,including 15 patients with HKαα/αα,three with HKαα/αα and β-thalassemia coinheritance,one with HKαα/-SEA,one with HKαα/-α4.2 andβ-thalassemia coinheritance,and one with-α3.7/αααanti-4.2 and β-thalassemia ***::αααanti-4.2 and HKααgenotypes of patients carrying-α3.7 need to be detected to reduce the misdiagnosis rate of patients carrying HKααand-α3.7/αααanti-4.2 *** accurate genetic counseling can be provided in the clinic using nested PCR combined