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Journal of Clinical Microbiology, December 1998, p. 3662-3669, Vol. 36, No. 12
0095-1137/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.

Sequence Variation within Three Important Cytomegalovirus Gene Regions in Isolates from Four Different Patient Populations

Benita Zweygberg Wirgart,1,* Maria Brytting,2 Annika Linde,2 Britta Wahren,2 and Lena Grillner1

Department of Clinical Microbiology, Section of Virology, Karolinska Hospital, S-171 76,1 and Swedish Institute for Infectious Disease Control, Microbiology and Tumorbiology Center, Karolinska Institute S-105 21,2 Stockholm, Sweden

Received 3 March 1998/Returned for modification 9 June 1998/Accepted 24 August 1998

We determined the nucleotide (nt) and amino acid (aa) heterogeneities of three distinct regions of the human cytomegalovirus (CMV) genome for 46 low-passage CMV isolates from four different patient populations (congenitally infected infants, children attending day-care centers, renal transplant recipients, and human immunodeficiency virus-infected individuals) and for two laboratory strains (CMV Ad169 and Towne). The gene regions for the major immediate-early (MIE) exon 4 gene (nt positions 1702 to 1982, aa positions 152 to 244), the DNA polymerase gene (nt positions 2797 to 3046, aa positions 713 to 795), and the glycoprotein B (gB) gene (nt positions 1698 to 1884, aa positions 567 to 628) were sequenced. The sequence information was used to design sets of nested PCR primers directed against the most highly conserved regions identified. MIE was the most variable gene region compared to the variability of the DNA polymerase and gB gene regions. Comparison of the sequences of all 46 isolates with that of Ad169 revealed nt and aa sequence homologies of 87.9 and 87.2%, respectively, within the MIE gene compared to 92.8 and 100% homologies, respectively, within the DNA polymerase gene and 93 and 95.2% homologies, respectively, within the gB gene. Within the MIE gene, compared to the Ad169 nt sequence the homology at the nt level among isolates obtained from children attending day-care centers was high (96.4%), while it was lower (90%) among isolates obtained from the other three patient populations. Preliminary results of a nested PCR with oligonucleotide primers selected from the DNA polymerase gene region with a low level of nt sequence variation indicates that primers selected from this region might be more powerful for use in PCR than primers selected from the MIE gene region.

* Corresponding author. Mailing address: Department of Clinical Microbiology, Section of Virology, Karolinska Hospital, S-171 76 Stockholm, Sweden. Phone: 46 8 51774925. Fax: 46 8 308099. E-mail: benita{at}mb.ks.se.

Journal of Clinical Microbiology, December 1998, p. 3662-3669, Vol. 36, No. 12
0095-1137/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.


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