A PCR based DNA hybridisation capture system for the detection of human cytomegalovirus. A comparative study with other identification methods.
A simple, sensitive and specific colourimetric hybridisation method for the detection of HCMV DNA in clinical specimens is described. This method combines a PCR assay with a sensitive sandwich hybridisation assay. It relies on the use of a specific capture probe linked covalently to polystyrene microplates and a specific polybiotinylated detection probe. Amplified DNA fragments, sandwiched between these two probes, are detected by an enzymatic colour reaction. This PCR-based colourimetric hybridisation method was compared with other known HCMV detection methods. Clinical specimens (n = 145, corresponding to 106 patients) were tested by both a nested PCR assay and this colourimetric hybridisation method; and by either the culture method or the pp65 antigenaemia test depending on the type of sample used. The results showed that the PCR-based hybridisation method has a specificity similar to tissue culture, known as the conventional gold standard method, and could be used for the examination of the clinical specimens.
Mansy
F
F
Université Libre de Bruxelles, Service de Génétique Appliquée, Nivelles, Belgium.
Brancart
F
F
Liesnard
C
C
Bollen
A
A
Godfroid
E
E
D000293:Adolescent; D000328:Adult; D000368:Aged; D000653:Amniotic Fluid / Q000737:chemistry / Q000821:virology; D002648:Child; D002675:Child, Preschool; D003587:Cytomegalovirus / Q000235:genetics*; D004279:DNA, Viral / Q000097:blood / Q000134:cerebrospinal fluid / Q000302:isolation & purification* / Q000652:urine; D019286:Endopeptidase K / Q000097:blood; D005260:Female; D006801:Humans; D008297:Male; D008875:Middle Aged; D009693:Nucleic Acid Hybridization / Q000379:methods*; D010920:Placenta / Q000737:chemistry; D016133:Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D014822:Vitreous Body / Q000737:chemistry
D004279:DNA, Viral; D019286:Endopeptidase K