Detection of human cytomegalovirus by slot-blot hybridisation assay employing oligo-primed 32P-labelled probe.
A 32P-labelled Hind III-0 DNA fragment (nine Kilobases; Kb) from human cytomegalovirus AD-169 (HCMV) was used in slot-blot hybridisation assay for the detection of HCMV in clinical samples. The results obtained with DNA hybridisation assay (DNA HA) were compared with virus isolation using conventional tube cell culture (CTC) and centrifugation vial culture (CVC), immunofluorescence (IF), and complement fixation test (CFT). Of 15 CTC-positive samples, 13 were positive with DNA HA (sensitivity 86.7%). Also, 14 additional samples were DNA HA-positive but CTC-negative. CVC and/or IF confirmed the diagnosis in nine of 14; the remaining five samples were from three patients who showed fourfold rising antibody titre by CFT. Although DNA HA using 32P-labelled probes is relatively cumbersome and expensive, it is a valuable test for quantitation of viral shedding in patients with HCMV infections who may benefit from antiviral therapy.
Agha
S A
SA
Department of Medical Microbiology, Charing Cross and Westminister Medical School, London, England.
Coleman
J C
JC
Selwyn
S
S
Mahmound
L A
LA
Abd-Elaal
A M
AM
Archard
L C
LC
D002478:Cells, Cultured; D003168:Complement Fixation Tests; D003587:Cytomegalovirus / Q000302:isolation & purification*; D003586:Cytomegalovirus Infections / Q000175:diagnosis*; D015342:DNA Probes; D004279:DNA, Viral / Q000032:analysis*; D005455:Fluorescent Antibody Technique; D006801:Humans; D009693:Nucleic Acid Hybridization; D010761:Phosphorus Radioisotopes; D012680:Sensitivity and Specificity
D015342:DNA Probes; D004279:DNA, Viral; D010761:Phosphorus Radioisotopes