Rapid detection of active cytomegalovirus infection by in situ polymerase chain reaction on MRC5 cells inoculated with blood specimens.
An in situ polymerase chain reaction was developed to amplify immediate early genes of human cytomegalovirus in cells cultured in a 96 well plate and infected with leukocytes. The technical parameters enabling optimal detection of the DNA sequences were defined. The key to this method is the fixation of cells, which facilitates the access of the PCR mixture into the cell nuclei and preserves cell morphology. Such a technique could have wide application for the detection and identification of other infectious viruses in cultured cells very early after inoculation of clinical samples.
Bettinger
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Laboratoire de Virologie, C.H.U. Saint-Jacques, Besançon, France.
Mougin
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Lab
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D001483:Base Sequence; D002460:Cell Line; D003587:Cytomegalovirus / Q000235:genetics* / Q000302:isolation & purification*; D003586:Cytomegalovirus Infections / Q000175:diagnosis* / Q000821:virology; D017931:DNA Primers / Q000235:genetics; D004279:DNA, Viral / Q000097:blood / Q000235:genetics; D005069:Evaluation Studies as Topic; D006801:Humans; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods*; D014773:Virology / Q000379:methods
D017931:DNA Primers; D004279:DNA, Viral