Glycoprotein B & H Genotype Of Human Cytomegalovirus Strains From Transplant Recipients

Human cytomegalovirus( HCMV)is responsible for server and often life threatening disease in immunocorapromised individuals especially in infants, AIDS patients and transplant recipients. Disease manifestations include pneumonitis, gastrointestinal tract lesions, retinitis and hepatitis. The glycoprotein B (gB) and glycoprotein H (gH) of HCMV are main target of adaptive response and have sequence of neutralizing sites and T cell epitops. According to the intragenic variability, HCMV strains can be classified into four gB genotypes and two gH genotypes. There has been evidence of differences in viral virulence; ethics and geography disperse among genotypes but has not been confirmed.In this study, gB & gH genotype has been detected in bone marrow and solid organ transplant recipients in order to learn (1) which gB-/gH- genotypes are represented among in bone marrow and solid organ transplant recipients in Zhejiang (2) is there a clustering of HCMV genotype restricted by geographic and ethnic origin or by the kind of transplantation.252 EDTA blood samples were obtained during routinescreening from 18 bone marrow transplant (BMT) ; 20 liver transplant (LT) ; 2 heart transplant (HT) and 57 renal transplant (RT) recipients who undertaken operation in the first affiliated hospital of school of medicine, Zhejiang university during December 1999 to September 2000. Genomic DNA was extracted from peripheral blood lymphocyte, which was prepared from the EDTA blood sample. According to the published HCMV gB and gH sequences, six pairs of primers were designed to amplify gBn(54-485bp), gBclv(1284-1600bp) and gH(-58~230bp) by the method of nest PCR. Sequencing was performed directly from the PCR-amplified gBn, gBclv and gH fragments after purification. Laboratory strain AD169 and Towne was sequenced repeatedly as control and sequence data were compared with published sequence. DNA sequence were analyzed and classified into different genotypes through phylogenetic analysis, x 2 test was used as statistics treatment by the software of SPSS-PC. The statistics significance was defined as p value<0. 05.From the study,we know that:1. Genotype information about gB and gH from clinical strains 3 gB genotypes and 2 gH genotypes were got from 23 sequence data responsible for 23 individuals. About gBn genotype, 3 (18.8%) had typel, 1(6.3%) had type 2, 11 (68.8%) had type 3, 1 (6. 3%) had mixture type of 1> 3, no type 4 was seen, gBn3 was most common seen( x '=12. 286,P=0.006) ; about gBclv genotype, 8 (57. 1?P had typel, 3 (21. 4%) had type 2, 3 (21. 4%) had type 3, no type 4 was seen; about gH genotype, 10 (50%) had gH typel, 6 (30%) had gH type 2, 4 (20%) had gH mixture type 1> 2. The distributions of gH and gBclv genotype were diversed.2. The distribution of gB and gH genotype among different kind of transplantation It seemed there was no difference between the type of transplantation and the genotype of strain.3. The accrodinary of gBclv and gBn genotype In 5 of 11 (45. 5%) strains, the gBn type appeared different from gBclv type. 4 appeared gBclvl-gBn3, 1 apperaed gBclv-2-gBnl, 3We can make conclusion as follows:1. There are gB genotype 1-3 and gH genotype 1-2 distribute in transplant recipients in Zhejiang province. gBn3 are most common seen.2. There is no difference between the genotype and the type of thransplantation.3. The intragenic variability within the gB gene, which due to homologous recombination occurs frequently.