Molecular Characterisation of Haemoglobin constant spring and Haemoglobin Quong Sze with a combine-amplification refractory mutation system

• Main Authors: Wee, Y.C., Tan, K.L., Chua, K.H., George, E., Tan, J.A.
• Format: Article
• Language: English
• Published: 2009
• Subjects: R Medicine
• Online Access: http://eprints.um.edu.my/3836/1/Molecular_characterisation_of_Haemoglobin_Constant_Spring_and_Haemoglobin_Quong_Sze_with_a_Combine-_Amplification_Refractory.pdf; http://eprints.um.edu.my/3836/

BACKGROUND: The interaction of the non-deletional α(+)-thalassaemia mutations Haemoglobin Constant Spring and Haemoglobin Quong Sze with the Southeast Asian double α-globin gene deletion results in non-deletional Haemoglobin H disease. Accurate detection of non-deletional Haemoglobin H disease, which is associated with severe phenotypes, is necessary as these mutations have been confirmed in the Malaysian population. METHODS: DNA from two families with Haemoglobin H disease was extracted from EDTA-anticoagulated whole blood and subjected to molecular analysis for α-thalassaemia. A duplex polymerase chain reaction was used to detect the Southeast Asian α-globin gene deletion. Polymerase chain reaction-restriction fragment length polymorphism analysis was then carried out to determine the presence of Haemoglobin Constant Spring and Haemoglobin Quong Sze. A combine-amplification refractory mutation system protocol was optimised and implemented for the rapid and specific molecular characterisation of Haemoglobin Constant Spring and Haemoglobin Quong Sze in a single polymerase chain reaction. RESULTS AND CONCLUSIONS: The combine-amplification refractory mutation system for Haemoglobin Constant Spring and Haemoglobin Quong Sze, together with the duplex polymerase chain reaction, provides accurate pre- and postnatal diagnosis of non-deletional Haemoglobin H disease and allows detailed genotype analyses using minimal quantities of DNA.