Rapid screening for sickle cell disease by polymerase chain reaction-high resolution melting analysis.
Each year, ~300,000 individuals with sickle cell disease (SCD), a hemoglobinopathy caused by β-globin gene mutation, are born, and >75% of those are in Africa. The present study examined 511 individuals on the island of Bioko (Equatorial Guinea) and attempted to establish a method for rapid sickle cell disease screening. Following DNA extraction and polymerase chain reaction (PCR) amplification, high resolution melting (HRM) analysis was used to assess the specificity of fluorescence signals of the PCR products and to differentiate various genotypes of these products. The analytical results of HRM were validated using DNA sequencing. By HRM analysis, 80 out of 511 samples were classified as hemoglobin S (Hb S) heterozygotes, while 431 out of 511 samples were classified as wild-type. No mutant homozygote was identified. DNA sequencing indicated that within the 431 wild-type samples as indicated by HRM analysis, one case was actually a Hb S heterozygote and another case was a rare hemoglobin S-C genotype (sickle-hemoglobin C disease). One out of 80 suspected Hb S heterozygotes as indicated by HRM was confirmed as wild-type by DNA sequencing and the results of residual 508 cases were consistent for HRM analysis and sequencing. In conclusion, HRM analysis is a simple, high-efficiency approach for Hb S screening and is useful for early diagnosis of SCD and particularly suitable for application in the African area.
Medical College of Shantou University, Shantou, Guangdong 515041, P.R. China.
Liang
L
Yue
Medical College of Shantou University, Shantou, Guangdong 515041, P.R. China.
Min
M
Lin
Laboratory Medical Center, Huizhou Central People's Hospital, Huizhou, Guangdong 516001, P.R. China.
Jiang-Tao
JT
Chen
Medical College of Shantou University, Shantou, Guangdong 515041, P.R. China.
Xiao-Fen
XF
Zhan
The Twenty-Fifth Bath of Chinese Medical Aid Team to the Republic of Equatorial Guinea, Guangzhou, Guangdong 511000, P.R. China.
De-Shang
DS
Zhong
Hospital Regional De Malabo, Malabo 999115, The Republic of Equatorial Guinea.
Santiago-M
SM
Monte-Nguba
The Twenty-Fifth Bath of Chinese Medical Aid Team to the Republic of Equatorial Guinea, Guangzhou, Guangdong 511000, P.R. China.
Pei-Fen
PF
Liu
The Twenty-Fifth Bath of Chinese Medical Aid Team to the Republic of Equatorial Guinea, Guangzhou, Guangdong 511000, P.R. China.
Xue-Fen
XF
Pan
The Twenty-Fifth Bath of Chinese Medical Aid Team to the Republic of Equatorial Guinea, Guangzhou, Guangdong 511000, P.R. China.
Jiang-Hua
JH
Huang
The Twenty-Fifth Bath of Chinese Medical Aid Team to the Republic of Equatorial Guinea, Guangzhou, Guangdong 511000, P.R. China.
Xi
X
Wang
Hospital Regional De Malabo, Malabo 999115, The Republic of Equatorial Guinea.
Juan Carlos Salas
JC
Ehapo
Hospital Regional De Malabo, Malabo 999115, The Republic of Equatorial Guinea.
Urbano Monsuy
UM
Eyi
Laboratory Medical Center, Chaozhou Central Hospital, Chaozhou, Guangdong 521021, P.R. China.
Hui-Tian
HT
Yang
Medical College of Shantou University, Shantou, Guangdong 515041, P.R. China.
Li-Ye
LY
Yang
D000293:Adolescent; D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D000755:Anemia, Sickle Cell / Q000175:diagnosis* / Q000453:epidemiology / Q000235:genetics; D002648:Child; D002675:Child, Preschool; D005260:Female; D006801:Humans; D007223:Infant; D008297:Male; D008875:Middle Aged; D016133:Polymerase Chain Reaction / Q000379:methods*; D055815:Young Adult