Diagnosis of thalassaemia by non-isotope detection of alpha/beta and zeta/alpha mRNA ratios.

The alpha/beta and zeta/alpha messenger RNA (mRNA) ratios in the thalassaemia syndromes were investigated by polymerase chain reaction (PCR) with silver staining of the PCR products. In this study we used the PCR to amplify cDNA copies of circulating erythroid cell mRNA in order to measure the relative amounts of alpha-, beta- and zeta-globin contained within. Quantitation was performed by scanning the silver stain of specific globin cDNA bands. We found that there were significant differences of alpha/beta-mRNA and zeta/alpha-mRNA in patients with Hb H disease and alpha-thalassaemia-1 compared to normal subjects. There was a marked increase in the alpha/beta-mRNA ratio but not in the zeta/alpha-mRNA ratio in patients with beta-thalassaemia. In two beta-thalassaemia cases abnormal increases of zeta-globin bands were noted and they were confirmed through DNA analysis to be combined with alpha-thalassaemia-1. This method provides a simple, rapid and non-radioactive approach to detect thalassaemia syndromes, and can help to screen cases of beta-thalassaemia with alpha-thalassaemia-1.