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Journal of Clinical Microbiology, July 2003, p. 2872-2877, Vol. 41, No. 7
0095-1137/03/$08.00+0     DOI: 10.1128/JCM.41.7.2872-2877.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Different Cytomegalovirus Glycoprotein B Genotype Distribution in Serum and Cerebrospinal Fluid Specimens Determined by a Novel Multiplex Nested PCR

David Tarragó,* Carmen Quereda,{dagger} and Antonio Tenorio

Microbiology and Parasitology Department and Infectious Diseases Department, Ramón y Cajal Hospital, 28034 Madrid, and Diagnostic Microbiology Department, Centro Nacional de Microbiología, Instituto de Salud Carlos III, 28220 Majadahonda, Madrid, Spain

Received 14 February 2002/ Returned for modification 7 April 2003/ Accepted 11 April 2003

A novel and highly sensitive multiplex nested PCR assay has been developed for the simultaneous glycoprotein B (gB) typing of four gB genotypes of human cytomegalovirus (CMV) directly from clinical specimens. Specifically, a pair of primers to conserved regions of all gB genotypes within the gB gene (gB and gpUL55) was used for primary amplification. A mixture of nested primers to specific and conserved regions of each gB genotype was used for a secondary PCR amplification, yielding amplicons of different sizes for each gB genotype that could easily be differentiated by agarose gel electrophoresis. A total of 40 of 44 serum specimens and 26 of 26 cerebrospinal fluid (CSF) samples, which had previously tested positive for CMV in 66 of 70 AIDS patients with different CMV disease conditions, were gB genotyped by this novel assay. Significant differences in glycoprotein B genotype distribution between serum and CSF specimens were found (P = 0.001). gB genotype 3 (gB3) and gB2 were the most prevalent genotypes in sera (42.5%) and CSF (38.5%), respectively. A different distribution was also observed when only patients with CMV retinitis were studied (P = 0.005), suggesting a gB2 neuron tropism. Neither CMV disease nor any particular clinical manifestation of CMV disease was associated with gB genotypes (P > 0.05). A high incidence of mixed infection with the gB1 and gB3 genotypes (27.5%) was detected in serum specimens, indicating that reinfection and reactivation are common traits in advanced AIDS patients.

* Corresponding author. Present address: Bacteriology Department, Centro Nacional de Microbiología, Instituto de Salud Carlos III, Ctra. Majadahonda-Pozuelo km 2, 28220 Majadahonda, Madrid, Spain. Phone: 34-915097901. Fax: 34-915097966. E-mail: davtarrago{at}isciii.es.

{dagger} Present address: Infectious Diseases Department, Ramón y Cajal Hospital, Madrid, Spain.

Journal of Clinical Microbiology, July 2003, p. 2872-2877, Vol. 41, No. 7
0095-1137/03/$08.00+0     DOI: 10.1128/JCM.41.7.2872-2877.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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