ATCC ATCC Logo 0

ATCC Minis Support VITEK 2 Quality Control Testing

an orange, sphere of Clostridium bacteria on a flat surface with lines dividing sections of it.

Demonstrating the use of ATCC Minis as quality control strains for microbial identification

Authors: Cara N. Wilder, PhD; Jennifer Murdock, MS; and Dev Mittar, PhD

This study will demonstrate the use of ATCC Minis as quality control strains for microbial identification on the VITEK 2 platform (bioMérieux), employing strains recommended for use with the Anaerobic and Corynebacteria (ANC), Bacillus (BCL), Corynebacteria (CBC), Gram-Positive (GP), Gram-Negative (GN), Neisseria-Haemophilus (NH), and Yeast (YST) Streamlined QC Sets as examples. 

Introduction

Commercial firms and research laboratories look to ATCC to provide top-quality microbial strains needed to maintain outstanding quality control programs for instruments, reagents, media, etc. To help aid in the rapid identification of microbial strains, ATCC now provides the same trusted quality control strains in a convenient, single-use, “mini” format that saves precious time and resources. ATCC Minis are a six pack of glass-free cryovials containing ready-to-use strains in glycerol stock, complete with peel-off labels for fast and reliable recordkeeping. Further, similar to their lyophilized counterparts, ATCC Minis are backed by polyphasic testing to establish identity, viability, and purity, and are maintained using a seed stock system to minimize subculturing. 

To provide convenient reference materials for fast, accurate microbial identification using the VITEK 2 system, ATCC has expanded its collection of quality control strains to include bacterial and fungal isolates in the ATCC Minis frozen format. In the following study, we demonstrate the application of ATCC Minis as quality control strains for microbial identification on the VITEK 2 platform using ANC, BCL, CBC, GP, GN, NH, and YST Streamlined QC Sets as examples. 

Materials and methods

ATCC Minis (Table 1) were thawed and cultured according to the recommended propagation procedure described on the product sheet. Following growth in broth, strains were streaked for isolation on the recommended agar medium and incubated under the appropriate temperature and atmospheric conditions until growth was established. To create the sample inoculum, isolated colonies were selected using a sterile cotton swab and suspended into 3.0 mL of 0.45% saline solution within a 12 mm x 75 mm polystyrene test tube. Inoculums were adjusted accordingly with the assistance of a calibrated DensiCHECK Plus (bioMérieux) to reach the McFarland Standard density range as recommended in the VITEK 2 Systems Product Information manual.1 The sample inoculum and associated VITEK 2 card were then inserted into a VITEK 2 Cassette and processed according to instructions found within the VITEK 2 Compact System and OBSERVA Computer System (bioMérieux) manuals. For this study, microbial identity was confirmed using the VITEK 2 ANC, BCL, CBC, GP, GN, NH, and YST cards, which provide species-level identification of anaerobes, bacilli, corynebacteria, Gram-positive bacteria, Gram-negative bacteria, Neisseria and Haemophilus strains, and yeasts, respectively.

 

Table 1. ATCC Minis

QC set ATCC number Species
VITEK 2 ANC Streamlined QC Set BAA-1296-MINI-PACK Bacteroides ovatus (ATCC BAA-1296)
  12464-MINI-PACK Clostridium septicum (ATCC 12464)
VITEK 2 BCL Streamlined QC Set 51663-MINI-PACK Brevibacillus agri (ATCC 51663)
VITEK 2 CBC Streamlined QC Set 43044-MINI-PACK Corynebacterium urealyticum (ATCC 43044)
  15829-MINI-PACK Microbacterium testaceum (ATCC 15829)
VITEK 2 GP Streamlined QC Set 700327-MINI-PACK Enterococcus casseliflavus (ATCC 700327)
  BAA-750-MINI-PACK Staphylococcus saprophyticus (ATCC BAA-750)
VITEK 2 GN Streamlined QC Set 700323-MINI-PACK Enterobacter hormaechei (ATCC 700323)
  17666-MINI-PACK Stenotrophomonas maltophilia (ATCC 17666)
VITEK 2 NH Streamlined QC Set BAA-1152-MINI-PACK Eikenella corrodens (ATCC BAA-1152)
VITEK 2 YST Streamlined QC Set 14053-MINI-PACK Candida albicans (ATCC 14053)

 

Results and discussion

As part of the microbial identification process, the VITEK 2 software compares the resulting biochemical reaction outcomes of organisms present in each sample to the expected set of results for organisms that have been previously identified by the database.1 These biochemical test outcomes are represented as a bionumber, a unique species identification pattern generated from the conversion of test results into positive or negative test probabilities. The identity percent probability is based on the similarity of the observed reactions compared to that of the expected reactions of an organism. Here, the possible range of percent probabilities for a single organism match is 85-99%, with values closer to 99% indicating a perfect match between the test reaction pattern and the unique reaction pattern of a known species.1

From this analysis, each of the ATCC Minis were identified using the appropriate VITEK 2 cards at a percent probability ranging from 91-99%, reflecting confidence levels of low, good, very good, and excellent (Table 2). Further, the results obtained using the ATCC Minis quality control strains corresponded with that of their lyophilized counterparts, indicating that the frozen format does not affect the biochemical phenotype of the strains (data not shown). 

BioMérieux selects quality control strains based on their biochemical characteristics. In some cases, these quality control strains exhibit expected biochemical results, but are unidentified or misidentified in the VITEK 2 system.1 As an example, the data for Brevibacillus agri ATCC 51633-MINI-PACK reflected an identity probability of 96% and a low confidence interval on the BCL card. This particular species belongs to a pseudoslashline taxa with Brevibacillus brevis. Thus, it is possible for the two species to exhibit the same biopattern, which results in low discrimination between B. agri and B. brevis

In some instances, VITEK 2 cards may be limited in the number of individual species within a given genera that can be identified.1 For example, the CBC card is capable of identifying Microbacterium spp. and Microbacterium lacticum, but not M. testaceum specifically. Accordingly, though the analysis of ATCC 15829-MINI-PACK, which is a strain of Microbacterium testaceum, exhibited a percent probability of 91% and a good confidence interval, it was identified only at the genus level. Therefore, in this instance, ATCC 15829-MINI-PACK performed as expected and was correctly identified within the identification constraints of the CBC card.1

 

Gloved hand holding a ATCC minis vial between forfinger and thumb

ATCC Minis are easy-to-use QC strains

See all our Minis


Table 2. VITEK 2 test results

ATCC number VITEK 2 card VITEK 2 lot number Analysis time (hours) Percent probability Bionumber Confidence level Species confirmed
BAA-1296-MINI-PACK ANC 244336210 6.25 99% 5057557176770 Excellent Bacteroides ovatus
12464-MINI-PACK ANC 244336210 6.25 99% 0000003042011 Excellent Clostridium septicum
51663-MINI-PACK BCL 239340320 14.25 96% 0727145601200401 Low Brevibacillus agri
43044-MINI-PACK CBC 246310810 8.25 99% 04000040000000 Excellent Corynebacterium urealyticum
15829-MINI-PACK CBC 246348420 8.00 91% 67577433625760 Good Microbacterium spp.
700327-MINI-PACK GP 242342010 4.75 93% 524213665777731 Very good Enterococcus casseliflavus
BAA-750-MINI-PACK GP 242351110 6.00 94% 030002056670131 Very good Staphylococcus saprophyticus
700323-MINI-PACK GN 241318040 5.00 98% 2623634553533010 Excellent Enterobacter hormaechei
17666-MINI-PACK GN 241318040 5.00 99% 1022103101540020 Excellent Stenotrophomonas maltophilia
BAA-1152-MINI-PACK NH 245341220 6.00 97% 0622002400 Excellent Eikenella corrodens
14053-MINI-PACK YST 243321210 18.50 99% 6102546065327771 Excellent Candida albicans

Conclusion

Overall, this study demonstrates that ATCC Minis exhibit results consistent with those expected for each respective VITEK 2 card and are identical to their lyophilized counterparts, making them ideal for use as quality control strains for the identification of unknown microbial strains using the VITEK 2 system.

Download a PDF of this application note

Download Now

References

  1. bioMérieux, Inc. VITEK 2 Systems Product Information, 2010.