Evaluation and comparison of PCR and hybridization methods for rapid detection of cytomegalovirus in clinical samples.
Rapid diagnosis of cytomegalovirus (CMV) infection may be obtained by molecular techniques, such as the polymerase chain reaction (PCR) and hybridization assays. The optimal technique to detect CMV in clinical samples was assessed. Two different PCR assays were used, targeting either the major immediate early 1 (MIE 1) or the HXLF 4 gene. The PCR products were detected by gel electrophoresis, dot blotting and an easy to use, rapid, solid phase hybridization assay, DNA enzyme immunoassay (DEIA). Standard tissue culture was also used. Cerebrospinal fluids (18), liver biopsies (9) from hepatic transplant recipients, amniotic fluids (7) from mothers with suspected peripartum infection, and samples (6) of miscellaneous origin (brain and fundus biopsy, pericardial and pleural fluid) were tested. Among the 40 samples, CMV was detected in 19 cases. Three were positive by both molecular techniques and tissue culture, 14 by molecular methods and 2 by culture. 16/19 or 9/19 CMV-positive samples were detected by PCR amplification of the HXLF 4 or MIE 1 gene, respectively and 14/16 HXLF 4-positive samples were detected using either dot-blot or DEIA, compared to 9/16 using gel electrophoresis. Thus, the most sensitive assays for the detection of CMV in clinical samples using the methods compared in the current study were PCR amplification of the HXLF 4 gene followed by dot-blot or DEIA hybridization.
Lévy
R
R
Laboratoire de Virologie, Faculté de Médecine Lyon Grange Blanche, France.
Najioullah
F
F
Thouvenot
D
D
Bosshard
S
S
Aymard
M
M
Lina
B
B
D000653:Amniotic Fluid / Q000821:virology; D001706:Biopsy; D001921:Brain / Q000821:virology; D002478:Cells, Cultured; D003587:Cytomegalovirus / Q000235:genetics* / Q000302:isolation & purification*; D003586:Cytomegalovirus Infections / Q000134:cerebrospinal fluid / Q000175:diagnosis*; D004279:DNA, Viral / Q000235:genetics* / Q000276:immunology / Q000302:isolation & purification*; D004587:Electrophoresis, Agar Gel; D005260:Female; D005347:Fibroblasts; D017781:Genes, Immediate-Early / Q000235:genetics; D006801:Humans; D007118:Immunoassay / Q000379:methods; D015151:Immunoblotting / Q000379:methods; D008099:Liver / Q000821:virology; D016031:Liver Transplantation; D009693:Nucleic Acid Hybridization / Q000379:methods*; D010490:Pericardial Effusion / Q000821:virology; D010996:Pleural Effusion / Q000821:virology; D016133:Polymerase Chain Reaction / Q000379:methods*; D011247:Pregnancy; D011251:Pregnancy Complications, Infectious / Q000821:virology; D012680:Sensitivity and Specificity
D004279:DNA, Viral