Abstract: Objective To develop a high resolution melting analysis (HRMA)-based genotypic testing for the detection of UL97 mutations of human cytomegalovirus (HCMV). Methods Wild-type UL97 and H520Q mutant were constructed. HRMA with unlabeled probe were used as genotyping method for the detection of H520Q mutation. Results The melting peaks directly from the PCR products were not able to distinguish wild-type from H520Q mutation, and the sensitivity and accuracy of HRMA were dramatically improved using unlabeled probe. HRMA with unlabeled probe successfully distinguished different contents of H520Q from wild-type. Conclusions HRMA with unlabeled probe proves to be a sensitive and cost-effective genotyping method for the detection of H520Q mutation.

Key words: human cytomegalovirus（HCMV）, UL97, high resolution melting analysis (HRMA)