Using the new CLSI M40-A2 Standard Coupled with Molecular Criteria to assess the performance of Sigma Transwab ® with Liquid Amies medium

Gizzie, Nina; Adukwu, Emmanuel

Abstract

Abstract: Background: Sigma-Transwab® is a liquid medium format transport swab designed for use on automated processing platforms. The PurFlock® swab is intended to improve absorption and release of the specimen. In 2014 the revised standard CLSI M40-A2 included new provisions for the evaluation of liquid medium transport swabs with novel bud types such as foam or flock. In this study, Sigma Transwab® PurFlock® (standard and minitip) and Sigma Transwab® (foam) was assessed to M40-A2, and a new step was added evaluating the ability to recover genomic DNA from the medium after the holding period.

Materials: The 10 bacterial strains specified in CLSI M40-A2 were used. Inoculated devices were held at controlled room temperature (RT) and 4°C. Viable counts were determined at 0, 24 and 48 h. Genomic DNA was extracted using the Qiagen DNeasy Blood & Tissue Kit, quantified by the Nanodrop 1000 and amplified by standard Polymerase Chain Reaction (PCR) and universal primers intended for the M40-A2 bacteria (16S ribosomal DNA).

Results: All three Sigma Transwabs® (foam and PurFlock® standard and minitip) were able to recover all specified strains at both 4°C and RT in accordance with M40-A2. Genomic DNA was successfully recovered after holding in the liquid Amies medium with high quality and yield after 24 hr. Data from the PCR analysis shows that the universal primer is able to amplify the 16S ribosomal DNA from the different bacterial genera.

Conclusion: All three Sigma Transwabs recovered all of the organisms specified in CLSI M40-A2. DNA integrity was maintained after holding in liquid Amies medium. The ability to amplify genomic DNA of the different genera on the M40-A2 standard using the universal primers can support quicker laboratory diagnosis, reduce confirmation times for bacterial infections and rapid identification tests.

• Presentation Conference Type: Poster
• Conference Name: Centre for Research in Biosciences (CRIB) Conference
• Start Date: Jan 15, 2016
• End Date: Jan 15, 2016
• Deposit Date: Jan 7, 2023
• Public URL: https://uwe-repository.worktribe.com/output/10307767
• Related Public URLs: https://www.uwe.ac.uk/research/centres-and-groups/biosciences/about-us