Evaluation of VIDAS UP Salmonella (SPT) assay for the detection of Salmonella in a variety of foods and environmental samples: collaborative study.

The VIDAS UP Salmonella (SPT) uses recombinant phage proteins to detect Salmonella species in human and animal food products and production environmental samples after 18-26 h of enrichment. The VIDAS SPT assay is performed with the automated VIDAS or mini-VIDAS instruments. The VIDAS SPT method was compared in a multilaboratory collaborative study to the U.S. Department of Agriculture/Food Safety and Inspection Service-Microbiology Laboratory Guidebook (USDA/FSIS-MLG) 4.05 (2011) Isolation and Identification of Salmonella from Meat, Poultry, Pasteurized Egg and Catfish Products reference method following the current AOAC guidelines. A total of 15 laboratories representing government, academia, and industry throughout the United States participated. One matrix, raw ground beef, was analyzed using two different test portion sizes, 25 and 375 g. Each test portion was artificially contaminated with Salmonella at three inoculation levels, an uninoculated control level (0 CFU/test portion), a low inoculum level (0.2-2 CFUltest portion), and a high inoculum level (2-5 CFU/test portion). In this study, 1656 unpaired replicate samples were analyzed. Of those unpaired replicates, 476 were presumptive positive by the VIDAS method, with 475 confirmed positive by the traditional confirmation procedures and 476 confirmed positive by an alternative confirmation procedure. There were 411 confirmed positive replicates by the USDA/FSIS-MLG reference method. Statistical analysis was conducted according to the probability of detection (POD). For the low-level 375 g test portions, the following dLPOD values, with 95% confidence intervals, were obtained: 0.01 (-0.12, +0.15) for samples confirmed following the traditional confirmation; 0.02 (-0.18, +0.2) for samples confirmed following traditional confirmation on IBISA and ASAP; and 0.03 (-0.18, +0.24) for samples confirmed following the alternative confirmation on IBISA and ASAP. For the low-level 25 g test portions, the following dLPOD values, with 95% confidence intervals, were obtained: 0.41, (0.32, +0.49) for samples confirmed following the traditional confirmation, the traditional confirmation on IBISA and ASAP, and the alternative confirmation on IBISA and ASAP. With 0.0 within the confidence intervals for the 375 g test portions, there was no statistically significant difference in the number of positive samples detected by the VIDAS SPT method and the USDA/FSIS-MLG method at the 0.05 level. For the 25 g test portions, a statistically significant difference was observed between the VIDAS SPT method and the reference method for the low inoculum level, where the VIDAS SPT method recovered a higher number of positive results than the reference method. It is recommended that the VIDAS SPT method with the optional ASAP and IBISA agar confirmation method be adopted for Official First Action status for the detection of Salmonella in a variety of foods and environmental samples.

Evaluation of the VITEK 2 gram positive (GP) microbial identification test card: collaborative study.

A collaborative study was conducted to evaluate the performance of the VITEK 2 Gram Positive (GP) identification card for use with the VITEK 2 automated microbial identification system. The GP test card is used in the identification of selected Gram positive organisms, including Listeria and Staphylococcus species. The VITEK 2 GP card is based on 43 biochemical tests measuring carbon source utilization, inhibition and resistance, and enzymatic activities. A total of 20 laboratories representing government, industry, and private testing laboratories throughout the United States participated. In this study, 720 Gram-positive inclusivity isolates were analyzed by the GP Identification method. Of the 720 well-characterized isolates, 714 were identified correctly, zero were misidentified, zero were unidentified, and six were not characterized as a Gram-positive organism by the VITEK 2 GP method. Additionally, 120 strains exclusive of Gram-positive organisms were screened by Gram stain. A total of 106 isolates were correctly excluded. Fourteen organisms were incorrectly characterized by Gram stain procedures, thus resulting in improper analysis and misidentification by VITEK GP. The VITEK 2 GP identification method is an acceptable automated method for the rapid identification of selected Gram-positive bacteria.

Evaluation of the VITEK 2 Gram-negative (GN) microbial identification test card: collaborative study.

A collaborative study was conducted to evaluate the performance of the VITEK 2 Gram-negative (GN) Identification card for use with the VITEK 2 automated microbial identification system. The GN test card is used in the identification of fermenting and nonfermenting Gram-negative bacilli, including the select agent organisms Brucella melitensis, Francisella tularensis, Burkholderia mallei, B. pseudomallei, and Yersinia pestis. The VITEK 2 GN card is based on 47 biochemical tests measuring carbon source utilization, inhibition and resistance, and enzymatic activities. A total of 20 laboratories representing government, industry, and private testing facilities throughout the United States participated. In this study, 720 Gram-negative inclusivity isolates were analyzed by the GN Identification method. Of the 720 well-characterized isolates, 707 were identified correctly, 0 were misidentified, 0 were unidentified, and 13 were not characterized as a Gram-negative organism. Additionally, 120 isolates exclusive of fermenting and nonfermenting Gram-negative bacilli were screened by Gram stain. A total of 117 isolates were correctly excluded. Three organisms were incorrectly characterized by Gram stain procedures, resulting in incorrect analysis and misidentification by VITEK 2 GN. The VITEK 2 GN identification method is an acceptable automated method for the rapid identification of Gram-negative bacteria.