Detection of multiple mycoplasma infection in cell cultures by PCR

A total of 301 cell cultures from 15 laboratories were monitored for mycoplasma (Mollicutes) using PCR and culture methodology. The infection was detected in the cell culture collection of 12 laboratories. PCR for Mollicutes detected these bacteria in 93 (30.9%) samples. Although the infection was c...

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Bibliographic Details
Published in:Brazilian journal of medical and biological research 2006-07, Vol.39 (7), p.907-914
Main Authors: Timenetsky, J, Santos, L M, Buzinhani, M, Mettifogo, E
Format: Article
Language:English
Subjects:
Base Sequence
BIOLOGY
Cell culture
Cells, Cultured - microbiology
DNA, Bacterial - analysis
Electrophoresis, Agar Gel
Humans
MEDICINE, RESEARCH & EXPERIMENTAL
Molecular Sequence Data
Multiple infection
Mycoplasma
Mycoplasma detection
Polymerase Chain Reaction
Tenericutes - classification
Tenericutes - genetics
Tenericutes - isolation & purification
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Summary:A total of 301 cell cultures from 15 laboratories were monitored for mycoplasma (Mollicutes) using PCR and culture methodology. The infection was detected in the cell culture collection of 12 laboratories. PCR for Mollicutes detected these bacteria in 93 (30.9%) samples. Although the infection was confirmed by culture for 69 (22.9%) samples, PCR with generic primers did not detect the infection in five (5.4%). Mycoplasma species were identified with specific primers in 91 (30.2%) of the 98 samples (32.6%) considered to be infected. Mycoplasma hyorhinis was detected in 63.3% of the infected samples, M. arginini in 59.2%, Acholeplasma laidlawii in 20.4%, M. fermentans in 14.3%, M. orale in 11.2%, and M. salivarium in 8.2%. Sixty (61.2%) samples were co-infected with more than one mycoplasma species. M. hyorhinis and M. arginini were the microorganisms most frequently found in combination, having been detected in 30 (30.6%) samples and other associations including up to four species were detected in 30 other samples. Failure of the treatments used to eliminate mycoplasmas from cell cultures might be explained by the occurrence of these multiple infections. The present results indicate that the sharing of non-certified cells among laboratories may disseminate mycoplasma in cell cultures.
ISSN:0100-879X
1414-431X
0100-879X
1414-431X
DOI:10.1590/S0100-879X2006000700009