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Mycoplasma detection in a historical arbovirus repository: Commercial kit comparison and implications for improved
Brandy J Russell1, Kalanthe Horiuchi1, Jason O Velez1
1Division of Vector-Borne Diseases, Centers for Disease Control and Prevention, Fort Collins, CO, United States.
Abstract:
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.
Insights
Mycoplasma contamination affects virus research. The PCR Mycoplasma Detection Kit (ABM) showed superior sensitivity and specificity compared to other kits for detecting contamination in arbovirus preparations.
Area of Science:
- Virology
- Microbiology
- Molecular Biology
Background:
- The Arbovirus Reference Collection (ARC) maintains viral isolates.
- Mycoplasma contamination can impact virus research and analysis.
- Routine quality control is essential for virus preparations.
Purpose of the Study:
- To evaluate the performance of three commercial mycoplasma detection kits.
- To compare these kits against a reference assay for arbovirus preparations.
Main Methods:
- Tested 744 virus preparations from the ARC using three kits: ABM, Sigma, and Lonza.
- Compared kit performance against the American Tissue Culture Collection (ATCC) reference assay.
- Assessed sensitivity, specificity, and false positive rates.
Main Results:
- The ABM kit demonstrated the highest sensitivity and specificity.
- The Sigma kit performed second best, followed by the Lonza kit.
- The Lonza kit showed increased false positives with samples passaged in suckling mouse.
Conclusions:
- The ABM kit is recommended for mycoplasma detection in arbovirus research.
- Mycoplasma contamination, once established, tends to persist in laboratory environments.
- Quality control testing is crucial for reliable arbovirus research.
