Mycoplasma detection in a historical arbovirus repository: Commercial kit comparison and implications for improved repository management

•Mycoplasma quality control testing is vital for continued maintenance of historical virus repositories.•Virus preparation (replication host) influences mycoplasma contamination result.•Specific mycoplasma species are maintained within a repository. The Centers for Disease Control and Prevention, Ar...

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Bibliographic Details
Published in:Journal of virological methods Vol. 276; p. 113769
Main Authors: Russell, Brandy J., Horiuchi, Kalanthe, Velez, Jason O., Goodman, Christin H., Johnson, Barbara W.
Format: Journal Article
Language:English
Published: Netherlands Elsevier B.V 01.02.2020
Subjects:
Animals
Arbovirus
Arboviruses
Biological Specimen Banks - standards
brain
cell culture
Cell Culture Techniques
Cell Line
Centers for Disease Control and Prevention
DNA, Bacterial - genetics
Humans
invertebrates
mice
Mycoplasma
Mycoplasma - isolation & purification
phenotype
polymerase chain reaction
Quality Control
Repository
RNA, Viral - genetics
suckling
tissue culture
unassigned viruses
viral growth
Virology - instrumentation
Virology - methods
Mycoplasma
Arbovirus
Repository
ISSN:0166-0934, 1879-0984, 1879-0984
Online Access:Get full text
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Summary:•Mycoplasma quality control testing is vital for continued maintenance of historical virus repositories.•Virus preparation (replication host) influences mycoplasma contamination result.•Specific mycoplasma species are maintained within a repository. The Centers for Disease Control and Prevention, Arbovirus Reference Collection (ARC) contains viral isolates from both environmental and human sources that are maintained in the laboratory through passage in suckling mouse brain and/or vertebrate and invertebrate cell culture. There has been increased concern regarding the effect of mycoplasma contamination on virus growth and its impact on research and phenotypic analysis. Therefore, quality control testing of virus preparations has become a routine part of the ARC quality assurance program. We compared the performance of three kits - the PCR Mycoplasma Detection Kit (ABM), the VenorGem Mycoplasma Detection Kit (Sigma), and the MycoAlert Mycoplasma Detection Kit (Lonza) - against a reference mycoplasma detection assay from the American Tissue Culture Collection (ATCC) using 744 virus preparations in the ARC, representing 721 unique viruses comprising twelve families and unclassified viruses. We found the ABM kit had the highest sensitivity and specificity, followed by the Sigma kit and Lonza kit, when compared to the ATCC kit. An increase in false positives was observed for the Lonza kit for preparations recently passaged in suckling mouse. Our data supports previously reported observations; that once introduced a specific species of mycoplasma is maintained within a lab.
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ISSN:0166-0934
1879-0984
1879-0984
DOI:10.1016/j.jviromet.2019.113769