Use of the polymerase chain reaction to analyse sequence variation within a major neutralizing epitope of glycoprotein B (gp58) in clinical isolates of human cytomegalovirus.
The heterogeneity of low passage human cytomegalovirus (HCMV) strains was determined by HindIII typing of 28 clinical isolates from transplant patients. These data have shown that, in general, each patient's strain has a unique restriction profile, usually comprising combinations of HindIII sites present in one or more of the tissue culture-adapted strains AD169, Towne and Davis. To map sequence changes in a more refined manner we performed detailed analyses of 33 low passage clinical isolates, including those aforementioned, analysing a sequence within glycoprotein B containing a major neutralizing epitope. A 149 bp sequence containing the epitope (amino acids 608 to 625) was amplified using the polymerase chain reaction, the products were cloned and their DNA sequence was determined. Comparison of the DNA and deduced amino acid sequences with those of HCMV strain AD169 revealed that there was a high degree of conservation of the epitope between the 33 clinical isolates. However 10 of the isolates possessed silent mutations and three isolates contained mutations producing amino acid changes within the neutralizing epitope. The possible functional significance of these changes is discussed.
Darlington
J
J
Division of Communicable Diseases, Royal Free Hospital School of Medicine, Hampstead, London, U.K.
Super
M
M
Patel
K
K
Grundy
J E
JE
Griffiths
P D
PD
Emery
V C
VC
D000595:Amino Acid Sequence; D000940:Antigenic Variation / Q000235:genetics*; D001483:Base Sequence; D015139:Blotting, Southern; D016025:Bone Transplantation; D002478:Cells, Cultured; D003587:Cytomegalovirus / Q000235:genetics* / Q000276:immunology; D004279:DNA, Viral; D000939:Epitopes / Q000235:genetics*; D006801:Humans; D016030:Kidney Transplantation; D008969:Molecular Sequence Data; D009500:Neutralization Tests; D016133:Polymerase Chain Reaction; D012150:Polymorphism, Restriction Fragment Length; D012689:Sequence Homology, Nucleic Acid; D014759:Viral Envelope Proteins / Q000235:genetics* / Q000276:immunology
D004279:DNA, Viral; D000939:Epitopes; D014759:Viral Envelope Proteins; C040495:glycoprotein B, Simplexvirus