ANALYSIS OF MESSENGER-RNA FROM RED-CELLS OF PATIENTS WITH THALASSEMIAAND HEMOGLOBIN-VARIANTS

During the past decade new procedures have been developed for the isol ation of RNA from a few mL of freshly collected blood. This material i s reverse transcribed and the resulting cDNA can be used for the deter mination of the ratios between different types of globin mRNA, namely alpha 2/alpha 1, alpha/zeta, alpha/beta, y/beta, beta(A)/beta(X), delt a beta(Lep)/beta and (G)y/(A)y. Details about these polymerase chain r eaction-based methods are reviewed, and information about their useful ness in studying alpha-thalassemia, beta-thalassemia, sickle cell anem ia and other beta-globin gene abnormalities, Hb Lepore heterozygosity, and heterozygosity for alpha 2- or alpha 1-globin gene mutations will be provided. The methods are also most useful in characterizing the m RNA types in single, in vitro cultured, BFU-E colonies; in colonies de rived from cells of a Hb S heterozygote; for instance, the beta(A)- an d beta(S)-mRNAs were present in all colonies and in about equal quanti ties, while many of those cells from a subject with a somatic cell mut ant (Hb Costa Rica) contained beta(A)-mRNA and no beta-Costa Rica mRNA , and only a few had both types. The techniques described have conside rable diagnostic value and offer a rather simple approach to the study of some of the listed diseases.