Establishing a rep-PCR DNA fingerprinting library for moulds
Abstract number: P728
Reece K., Atkins E., Ross T., Abedi S., Smith C., Dutch W., Wise M., Embry J., Frye S., Healy M.
Objectives: Discrimination among moulds is often difficult. The advances in many areas in microbiology are limited when looking at moulds. Morpholological characterisation and staining techniques are still very common. Sequencing and real-time PCR can be used but additional technologies are needed for investigation of moulds. Previous studies have shown the promise for the use if rep-PCR DNA fingerprinting as a molecular method to characterise some moulds. This study reports the compilation of an expanded Mould database as an additional tool that can be applied to mould characterisation and subspecies discrimination.
Methods: A total of 250 previously characterised moulds (including strains from 30 Genus/species) obtained from environmental and clinical sites and ATCC reference strains were cultured. The isolates were extracted and then amplified using the Mold Kit for DNA fingerprinting. The amplified product was separated using a microfluidic based detection. The resulting data was analysed using dendrogram analysis and a Mould library was created using the DiversiLab Software. An additional 50 mould isolates were processed and compared to the library for validation.
Results: The resulting fingerprints from the first 250 characterised isolates grouped into multiple clusters. No different Genus/species showed the same fingerprint pattern. Some species had more than one fingerprint pattern, or cluster, indicating discrimination to the subspecies and/or strain level. Many of the 50 isolates used for validation showed matches to the library. Some isolates formed new clusters but no false positive groupings are seen.
Conclusions: These data suggest that a characterised library may prove useful to provide additional information or confirmation when using DNA fingerprinting for moulds. The DiversiLab System shows promise as a tool to aid in surveillance and identification of hospital acquired infections and epidemiological studies.
|Session name:||18th European Congress of Clinical Microbiology and Infectious Diseases|
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