Some features and advantages of TAAG F Pathogens PCR kits
for pathogen detection

Multiplex technology

Identify up to four different pathogens simultaneously in a single PCR reaction

28hr

Optimized workflow

Reduce your operation time by obtaining results in 28 hours, enrichment included

100%

Integrated systems

Automatic results interpretation and upload to TxA Lab and TxA Dashboard platform

100%

Reliable

Along with pathogen detection, the inclusion of an internal control validates your PCR run and your negative results.

Cost-effective

Use fewer supplies, optimize your equipment and reduce processing time up to four times

TAGN01009

TAAG F Pathogens Real-Time PCR Kits

The technology behind TAAG F Pathogens real-time PCR kits allows the simultaneous detection of the most important pathogens of food industry: Salmonella spp, Staphylococcus aureus, Escherichia coli, Escherichia coli O157:H7 and Listeria monocytogenes, in food and surfaces samples, in a single real-time PCR reaction.

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It’s simple: reduce your costs, increase your productivity

Improving food safety has never been easier. Our multiplex real-time PCR kits are designed to reduce your costs up to four times in every step of your process, by optimizing both your workflow and resources, allowing you to get faster results in less time. For a laboratory analyzing around 50 samples per day, for 4 pathogens, e.g. Listeria monocytogenes, Salmonella spp, Escherichia coli and Staphylococcus aureus, here’s what the numbers look like:

less time to obtain results

29%

of cost reduction in the detection of 4 pathogens

TAAG F Pathogens real-time PCR kits are multiplex PCR tests for the detection of the main pathogens that affect food industry, in a single reaction. This kit contains a real-time PCR mix, which has the necessary primers and reagents for the simultaneous detection up to four microorganismos, such as Escherichia coli, Escherichia coli O157:H7, Listeria monocytogenes, Staphylococcus aureus and Salmonella spp, as well as an internal control, to ensure the reliability of your process.

KIT PERFORMANCE

Inclusivity

in silico inclusivity was was performed by BLAST analysis of the primer sequences for bacterial detection present in TAAG F Pathogens real-time PCR kits, against all available genome sequences published in NCBI of Listeria monocytogenes, Salmonella spp, Escherichia coli and Staphylococcus aureus. Table 1 summarizes the number of analyzed sequences for each microorganism, and the inclusivity percentage of TAAG F Pathogens real-time PCR kits.

**Table 1.** Experimental selectivity analysis
Microorganism	Analyzed sequences	Inclusive percentage
Listeria monocytogenes	938	100%
Salmonella spp	643	100%
Escherichia coli	264	100%
Staphylococcus aureus	776	100%

Selectivity

in silico selectivity was performed by BLAST analysis of the primer sequences for bacterial detection present in TAAG F Pathogens real-time PCR kits against the non-redundant genome database from NCBI. Listeria monocytogenes, Salmonella spp, Escherichia coli and Staphylococcus aureus were excluded from the analysis. No match was found for each set of primers against any other relevant organism that could lead to false-positive results, thus demonstrating in silico selectivity of TAAG F Pathogens real-time PCR kits (data not shown).

**Table 2.** in silico inclusivity analysis
Microorganism	Experimental selectivity - Pure strains	Experimental selectivity - Food strains	Experimental selectivity - Surface samples	Percentage of selectivity
Listeria monocytogenes	72	3408	2312	100%
Salmonella spp	103	9212	2272	100%
Escherichia coli	93	516	2276	100%
Staphylococcus aureus	95	508	2048	100%

Results from the analysis of over 2000 microorganism sequences show that TAAG F Pathogens kits are 100% inclusive, which completely reduces the obtention of false negative results.

Experimental inclusivity was performed by detecting bacterial DNA using TAAG F Pathogens kits from pure strains of Listeria monocytogenes (42 strains), Salmonella spp (9 strains), Escherichia coli (19 strains), and Staphylococcus aureus (17 strains). As a result, all 87 pure strains were sucessfully detected our TAAG F Pathogens kits.

Limit of Detection

To determine the limit of detection of our real-time PCR kits, DNA from Listeria monocytogenes, Salmonella spp, Staphylococcus aureus y Escherichia coli of known concentrations were diluted and analyzed, which allowed the determination of the minimal number of equivalent genomic copies (CGE) our kits are able to detect. Results are considered to be accepted if it is possible to detect 1000 copies of the target genes, equivalent to 10^3 genomic copies of Listeria monocytogenes, Salmonella spp, Staphylococcus aureus and Escherichia coli, post-enrichment. Results are summarized in table 3.

**Table 3.** Limit of detection of TAAG F Pathogens real-time PCR kits, performed by analysis of seriated dilutions of bacterial DNA.
	Food Total
	10^5 CGE	10^4 CGE	10^3 CGE	10^2 CGE	10^1 CGE	10^0 CGE
Listeria monocytogenes	+	+	+	+	+	-
Salmonella spp	+	+	+	+	+	-
Staphylococcus aureus	+	+	+	+	-	-
Escherichia coli	+	+	+	+	-	-

The evaluated experimental conditions demonstrate our kit’s sensitivity, which is able to identify very low amounts of DNA in the samples.

Limit of Detection in different food samples

The sensitivity of the kit was evaluated in artificially contaminated food samples with a low amount of microorganism to be detected ( <5 UFC in 25 g of food sample). Samples were analyzed in three independent experiments, and results were informed as accepted when samples were negative for the non-inoculated sample, and the positive controls in each case showed positive results. Results are summarized in table 4.

**Table 4.** Limit of detection of TAAG F Pathogens real-time PCR kits, performed in artificially contaminated sapmles.
Poultry
		L. monocytogenes	Salmonella spp.	E. coli	S. aureus
Raw	Whole chicken	+	+	+	+
	Turkey breast	+	+	+	+
	Chicken fillet	+	+	+	+
	Turkey ribs	+	+	+	+
Heat-processed	Roasted turkey breast	+	+	+	+
	Roasted turkey legs	+	+	+	+
	Whole roasted chicken	+	+	+	+
	Roasted chicken breast	+	+	+	+
Frozen	Frozen chicken fillets	+	+	+	+
Frozen	Whole frozen turkey	+	+	+	+
Meats
		L. monocytogenes	Salmonella spp.	E. coli	S. aureus
Raw	Ground beef	+	+	+	+
	Minced beef	+	+	+	+
	Beef tenderloin	+	+	+	+
	Pork tenderloin	+	+	+	+
	Ground pork	+	+	+	+
Heat-processed	Roasted meat and rice	+	+	+	+
Heat-processed	Roasted chicken and mashed potatoes	+	+	+	+
Frozen	Frozen pork meat	+	+	+	+
Fermented / cured	Saussages	+	+	+	+
	Bacon	+	+	+	+
	Ham	+	+	+	+
Fruit and vegetables
		L. monocytogenes	Salmonella spp.	E. coli	S. aureus
Raw	Fruit Mix	+	+	+	+
Raw	Cabbage, broccoli and culiflower mixed salad	+	+	+	+
Heat-processed	Pasteurized apple juice	+	+	+	+
Frozen	Frozen broad beans	+	+	+	+
	Frozen corn	+	+	+	+
	Frozen broccoli	+	+	+	+
Fermented	Fermented cabbage	+	+	+	+
Cured / salted	Pickles	+	+	+	+
Other	Mushrooms	+	+	+	+
Fish and sea-products
		L. monocytogenes	Salmonella spp.	E. coli	S. aureus
Raw	Fresh salmon	+	+	+	+
Heat-processed	Seafood assortment	+	+	+	+
Frozen	Frozen tilapia	+	+	+	+
Smoked	Smoked salmon	+	+	+	+
Dairy
		L. monocytogenes	Salmonella spp.	E. coli	S. aureus
Raw	Raw milk	+	+	+	+
Heat-processed	Pasteurized milk	+	+	+	+
Frozen	Chocolate ice-cream	+	+	+	+
Fermented	Strawberry yoghurt	+	+	+	+
Dryed	Powdered milk 26% fat	+	+	+	+
Chocolate and bakery
		L. monocytogenes	Salmonella spp.	E. coli	S. aureus
Low humidity	Chocolate 85% Cocoa	+	+	+	+
Dryed	Cake mix	+	+	+	+
Other products
		L. monocytogenes	Salmonella spp.	E. coli	S. aureus
Spices	Oregano	+	+	+	+
Sauces	Mole sauce	+	+	+	+
Eggs	Fresh eggs	+	+	+	+
Manipulators and surfaces
		L. monocytogenes	Salmonella spp.	E. coli	S. aureus
Post-sanitization	Chicken clamp	+	+	+	+
Post-sanitization	Vegetable cutting board	+	+	+	+
In use	Chicken transport tray	+	+	+	+
In use	Vegetable transport tray	+	+	+	+

Analysis of frozen specimens from frozen fruits, vegetables and meat

914 total frozen samples were analyzed using our real-time PCR kit to detect any of the four microorganisms (Listeria monocytogenes, Salmonella spp, Escherichia coli and Staphylococcus aureus). 4.8% of the samples were positive for at least one of the aforemetioned bacteria. Analyzed frozen food samples were:

Frozen vegetables:

Broccoli.

Cauliflower.

Corn.

Chickpeas.

Broad beans.

Green beans.

Mixed salad.
Frozen fruits:

Strawberries.

Raspberries.

Papayas.

Custard apples.

Blueberries.
Frozen meat.

Frozen beef.

Frozen chicken.

Frozen pork.
Frozen fish.

Pippin.

Conger.

Salmon.

Targets of detection	Listeria monocytogenes, Salmonella spp, Escherichia coli, Escherichia coli O157:H7, Staphylococcus aureus
Technique	Real-time PCR
Detection technology	Fluorescence dyes
Channels	FAM
Controls	Positive controls for Listeria monocytogenes, Salmonella spp, Escherichia coli, Escherichia coli O157:H7, Staphylococcus aureus; Negative PCR control, and an internal control of PCR inhibition to validate negative results.
Type sample	Food, surface and water samples
Initial nucleic acid	DNA

User Manual	Manual kit TAAG F41 Pathogens.
MSDS	MSDS kit TAAG F41 Pathogens.
Technical Specifications	TAAG F41 Pathogens Kit.
Validations	Validations FOOD Total Kit.

TAGN01001, TAGN01003, TAGN01009

TAAG F Pathogens kits for pathogen detection

Multiplex real-time PCR for the detection of foodborne pathogens,
conceived to optimize your workflow and maximize your productivity.

Some features and advantages of TAAG F Pathogens PCR kits
for pathogen detection

Multiplex technology

Optimized workflow

Integrated systems

Reliable

Cost-effective

TAGN01009

TAAG F Pathogens Real-Time PCR Kits

It’s simple: reduce your costs, increase your productivity

KIT PERFORMANCE

TAAG Genetics

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TAGN01001, TAGN01003, TAGN01009

TAAG F Pathogens kits for pathogen detection

Multiplex real-time PCR for the detection of foodborne pathogens,conceived to optimize your workflow and maximize your productivity.

Some features and advantages of TAAG F Pathogens PCR kits for pathogen detection

Multiplex technology

Optimized workflow

Integrated systems

Reliable

Cost-effective

TAGN01009

TAAG F Pathogens Real-Time PCR Kits

It’s simple: reduce your costs, increase your productivity

KIT PERFORMANCE

Choose your region

Choose your language

Multiplex real-time PCR for the detection of foodborne pathogens,
conceived to optimize your workflow and maximize your productivity.

Some features and advantages of TAAG F Pathogens PCR kits
for pathogen detection