Molecular prenatal diagnosis of alpha-thalassemia using real-time and multiplex polymerase chain reaction methods.
Molecular analysis of two fetuses at high risk of alpha-thalassemia (alpha-thal), and their family members, was performed using real-time polymerase chain reaction (PCR) with SYBR Green 1 (SYBR-PCR) dye combined with dissociation curve analysis and multiplex PCR (m-PCR) and DNA sequencing techniques. The genotype of the fetus from one family was --SEA/--SEA (Southeast Asian deletion), which produces hydrops fetalis syndrome. The genotype of the parents was --SEA/alphaalpha. A boy with Hb H disease and his sibling fetus from the other family had the genotype --SEA/alphaCSalpha [the Hb Constant Spring (CS) mutation: alpha142, Term-->Gln (TAA>CAA in alpha2)] and alphaalpha/alphaalpha (normal), respectively. The diagnosis, based on SYBR-PCR combined with dissociation curve analysis, was in agreement with the results from the m-PCR method. This indicates that these are alternative and reliable assays for the molecular diagnosis of deletional alpha-thal.
Liu
Jing-Zhong
JZ
Basic Medical Research Center, Beijing Chaoyang Hospital, Capital Medical University, Beijing, China. liujingzhong@hotmail.com
Yan
Mei
M
Wang
Li-Rong
LR
Wang
Zhan-Yong
ZY
Fan
Yin-Ping
YP
Xiao
Bai
B
Zhou
Yan
Y
D000328:Adult; D002675:Child, Preschool; D005260:Female; D005333:Fetus / Q000473:pathology; D005820:Genetic Testing / Q000379:methods*; D005838:Genotype; D006801:Humans; D008297:Male; D010290:Parents; D016133:Polymerase Chain Reaction / Q000379:methods*; D011247:Pregnancy; D011296:Prenatal Diagnosis / Q000379:methods*; D017085:alpha-Thalassemia / Q000175:diagnosis* / Q000235:genetics