Glycoprotein B genotype of human cytomegalovirus analysis by nPCR-RFLP / 中华传染病杂志

ABSTRACT

Objective Aim to set up the method of glycoprotein B genotype of human cytomegalovirus (HCMV) analysis by using nested polymerase chain reaction(nPCR) and restriction fragment length polymorphism(RFLP). Methods Eleven blood samples and twenty-three urine samples were obtained from thirty-four HCMV-infected patients. A fragment of gB gene was amplified by nPCR. HCMV gB genotyping was carried out by RFLP, and the amplified DNA fragments were verified by DNA sequencing. Results Of the 34 patients, gB type Ⅰ was found in 13 patients, gB type Ⅱ in 12 patients, gB type Ⅲ in 9 patients, and none had the gB type Ⅳ sequence. The similarities of PCR products of HCMV gB Ⅰ, Ⅱ and Ⅲ amplified compared with the sequences of prototype strains in GenBank were 98.1%～99.6%, 98.9%～100%, 97.3%～98.9%. Conclusions The nPCR assay developed in this study was sensitive and specific for detection of HCMV, and RFLP analysis of HCMV gB genotype was definite and reliable.