PCR SSCP and silver staining technique in screening insulin receptor gene mutation

Objective To establish polymerase chain reaction single strand conformation polymorphism(PCR SSCP) and silver staining technique to screen mutation of insulin receptor gene. Methods 89 patients with NIDDM were collected.Exon 17 21 of insulin receptor gene were amplified by PCR and screened by SSCP electrophoresis.After silver staining,the mutation samples was sequenced.Resulets Eleven variant sequences in exon 17 and 1 in exon 20 were detected. Direct sequence analysis of exon 17 showed that homozygous and heterozygous nonsense mutations were silent polymorphism at position 1058 (CAC→CAT).Conclusion The PCR SSCP combining with silver staining technique is simple, economic, and sensitive.