Abstract
A sensitive semi-nested polymerase chain reaction (PCR) was established which allows rapid identification of human cytomegalovirus strains directly on clinical specimens, thereby permitting virus isolation and propagation on cell cultures to be avoided. The assay is based on restriction analysis of PCR products derived from the polymorphic UL10-UL13 region of the human cytomegalovirus genome. The method was evaluated using clinical samples from 23 subjects comprising 16 breast-feeding mothers and seven bone marrow transplantation recipients. For eight mothers, postnatal virus transmission to their offspring via breast milk was studied. Interestingly, for one mother-infant pair, a double infection with two distinct human cytomegalovirus strains could be demonstrated. Stepwise digestion with different restriction enzymes raised the possibility of detecting different strains almost twofold compared to analysis with only one enzyme. This assay is a practical tool for monitoring human cytomegalo-virus transmission in various clinical settings.
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References
Chou S: Acquisition of donor strains of cytomegalovirus by renal-transplant recipients. New England Journal of Medicine (1986) 314:1418–1423
Huang E, Alford C, Reynolds DW, Stagno S, Pass RF: Molecular epidemiology of cytomegalovirus infections in women and their infants. New England Journal of Medicine (1980) 303:958–962
Spector SA, Neuman TA, Hirata KK: Rapid determination of molecular relatedness of isolates of human cytomegalovirus. Journal of Infectious Diseases (1985) 152:755–759
Zaia JA, Gallez-Hawkins G, Churchill MA, Morton-Blackshere A, Pande H, Adler S, Schmidt GM, Forman SJ: Comparative analysis of human cytomegalovirusα-sequence in multiple clinical isolates by using polymerase chain reaction and restriction fragment length polymorphism assay. Journal of Clinical Microbiology (1990) 28:2602–2607
Watanabe S, Shinkai M, Hitomi S, Kozuka H, Kimura S, Shimada K, Hondo R, Yamaguchi N: A polymorphic region of the human cytomegalovirus genome encoding putative glycoproteins. Archives of Virology (1994) 137:117–121
Hirt B: Selective extraction of polyoma DNA from infected mouse cell cultures. Journal of Molecular Biology (1967) 26:365–369
Hamprecht K, Vochem M, Baumeister A, Boniek M, Speer CP, Jahn G: Detection of cytomegaloviral DNA in human milk cells and cell free milk whey by nested PCR. Journal of Virological Methods (1998) 70:167–176
Vochem M, Hamprecht K, Jahn G, Speer CP: Transmission of cytomegalovirus to preterm infants through breast milk. Pediatric Infectious Disease Journal (1998) 17:53–58
Chou S, Dennison KM: Analysis of interstrain variation in cytomegalovirus glycoprotein B sequences encoding neutralization-related epitopes. Journal of Infectious Diseases (1991) 163:1229–1234
Smith IL, Shinkai M, Freeman WR, Spector SA: Polyradiculopathy associated with ganciclovir-resistant cytomegalovirus in an AIDS patient: phenotypic and genotypic characterization of sequential virus isolates. Journal of Infectious Diseases (1996) 173:481–484
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Prix, L., Kuner, R., Speer, C.P. et al. Evaluation of restriction fragment length polymorphism analysis of the UL10-UL13 genomic region for rapid identification of human cytomegalovirus strains. Eur. J. Clin. Microbiol. Infect. Dis. 17, 525–528 (1998). https://doi.org/10.1007/BF01691140
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DOI: https://doi.org/10.1007/BF01691140