Detection of cell culture mycoplasmas by a genetic probe.
The utility of a genetic probe prepared against mycoplasmal ribosomal RNA was evaluated. The probe consisted of a tritiated DNA probe prepared with reverse transcriptase against Mycoplasma and Acholeplasma. Using a batch process without Southern blots and autoradiography, the probe assay detected mycoplasma infection in 27 of 52 cell cultures, an incidence of 51.9%. No false-positives were detected. Two false-negatives occurred; these were unusual mycoplasma infections of cell cultures grown in serum-free media. The sensitivity of the probe varied with mycoplasma species, from 9.3 X 10(3) for M. arginini to 2.6 X 10(6) for M. orale. The assay is rapid, taking only 2-3 h.
G J
GJ
McGarrity
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H
Kotani
D000127:Acholeplasma / Q000235:genetics / Q000302:isolation & purification*; D002478:Cells, Cultured / Q000382:microbiology*; D004247:DNA; D005188:False Negative Reactions; D005189:False Positive Reactions; D009174:Mycoplasma / Q000235:genetics / Q000302:isolation & purification*; D009693:Nucleic Acid Hybridization; D012329:RNA, Bacterial / Q000032:analysis*; D012335:RNA, Ribosomal / Q000032:analysis*
D012329:RNA, Bacterial; D012335:RNA, Ribosomal; D004247:DNA