title: Development and evaluation of a reverse dot blot assay for the simultaneous detection of common alpha and beta thalassemia in Chinese.
abstract: Thalassemia is the commonest inherited autosomal recessive disorders of hemoglobin in southern China. We developed and evaluated a reverse dot blot (RDB) assay combined with flow-through hybridization technology platform for the rapid and simultaneous identification of 5 types of α-thalassemia and 16 types of β-thalassemia common in Chinese. Reliable genotyping of wild-type and thalassemic genomic DNA samples was achieved by means of a gene chip on which allele-specific oligonucleotide probes were immobilized on a nylon membrane. This method involved two multiplex PCR amplification systems of α-thalassemia and β-thalassemia and one time of hybridization. The whole procedure starting from blood sampling to the identification of thalassemia genotype required less than 4h. The diagnostic reliability of this reverse dot blot assay was evaluated on 427 samples (387 cases of thalassemia and 40 healthy persons) by using direct DNA sequence analysis and gap-PCR in a blind study. These samples included 377 cases of blood, 7 cases of amniotic fluid, 18 cases of chorionic villus, and 25 cases of cord blood. The RDB gene chip was in complete concordance with the reference method. The reverse dot blot assay was a simple, rapid, accurate, and cost-effective method to identify common thalassemia genotypes in the Chinese population.
authors.affiliation: Laboratory Medical Center, Chaozhou Central Hospital Affiliated to Southern Medical University, Chaozhou, Guangdong Province, PR China.
authors.forename: Min
authors.initials: M
authors.lastname: Lin
authors.forename: Juan-Juan
authors.initials: JJ
authors.lastname: Zhu
authors.forename: Qian
authors.initials: Q
authors.lastname: Wang
authors.forename: Long-Xu
authors.initials: LX
authors.lastname: Xie
authors.forename: Min
authors.initials: M
authors.lastname: Lu
authors.forename: Jun-Li
authors.initials: JL
authors.lastname: Wang
authors.forename: Chun-Fang
authors.initials: CF
authors.lastname: Wang
authors.forename: Tian-Yu
authors.initials: TY
authors.lastname: Zhong
authors.forename: Lei
authors.initials: L
authors.lastname: Zheng
authors.forename: Mei-Chen
authors.initials: MC
authors.lastname: Pan
authors.forename: Jiao-Ren
authors.initials: JR
authors.lastname: Wu
authors.forename: Ying-Fang
authors.initials: YF
authors.lastname: Wen
authors.forename: Gui-Rong
authors.initials: GR
authors.lastname: Liu
authors.forename: Xiao-Fen
authors.initials: XF
authors.lastname: Zhan
authors.forename: Fen
authors.initials: F
authors.lastname: Lin
authors.forename: Li-Ye
authors.initials: LY
authors.lastname: Yang
mesh_terms: D044466:Asian People / Q000235:genetics; D001483:Base Sequence; D002681:China; D006801:Humans; D060885:Multiplex Polymerase Chain Reaction / Q000379:methods; D009154:Mutation; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D055542:alpha-Globins / Q000235:genetics; D017085:alpha-Thalassemia / Q000175:diagnosis* / Q000235:genetics; D055544:beta-Globins / Q000235:genetics; D017086:beta-Thalassemia / Q000175:diagnosis* / Q000235:genetics
chemical_list: D055542:alpha-Globins; D055544:beta-Globins