Part of the TAAG technology pillar. It complements Ampliora, Elevia and Specio and how to choose a TAAG kit.
Table of contents
A technical data sheet is a scope contract
Before buying or implementing a PCR kit, the laboratory receives a technical data sheet with a dozen fields: format, contents, fluorescence channels, times, sensitivity, matrices, compatible instruments, storage and, sometimes, a certification. It describes the conditions under which the manufacturer demonstrated that the method works. Outside those conditions, the result is no longer backed by the validation.
This article goes through those fields with real examples from the TAAG catalog, applicable to any supplier's data sheet. The differences between product lines are covered in Ampliora, Elevia and Specio and the choice by industry in how to choose a TAAG kit.
Format and contents: what comes in the box
On the data sheet for Ampliora 1.1 Salmonella spp. (code V-SF97), the declared format is SPID, for 96 reactions, and the contents are 12 pre-loaded PCR strips, one 100 µL positive control tube and one 352 µL negative control tube. Specio 4.1 (V-SF95) comes in 480 reactions, with 60 pre-loaded strips and five tubes of each control.
Two things are worth checking:
- What is included and what is not. What is not in the box (enrichment medium, extraction kit) is still part of the method. The Ampliora 1.1 data sheet lists as related products the Augmentis 14 Universal Gram negative medium, the Nucleia 2 Tez-Q Plus extraction kit and the Collectio 1 NeutroSampling sampling kit. Replacing them changes the protocol that was evaluated.
- The controls. They tell you whether the run was valid, as explained in PCR controls.
Detection chemistry and fluorescence channels
Probes or intercalating fluorophore
The chemistry field indicates how the signal is generated. Ampliora data sheets declare fluorescent probes: a labelled oligonucleotide that hybridizes within the target sequence and generates signal as the product accumulates. Holland and colleagues described this kind of specific detection during amplification in 1991. Specio data sheets, by contrast, declare an intercalating fluorophore, which binds to any double-stranded DNA. That is why those kits add a melting curve analysis: according to Ririe, Rasmussen and Wittwer (1997), the shape and position of that curve depend on the GC content, length and sequence of the product, and make it possible to distinguish products whose melting temperatures differ by less than 2 °C.
What FAM, HEX, ROX or Cy5 mean
They are the names of the fluorophores, and each one is read in a different optical channel of the thermal cycler. A probe-based kit assigns one channel to each target and, when included, another to a control. Some literal examples from the catalog:
- Ampliora 1.1 Salmonella spp.: FAM and HEX.
- Ampliora 3.5 (Salmonella spp., L. monocytogenes and Listeria spp.): FAM, HEX, ROX and Cy5.
- Ampliora 3.9 (STEC, E. coli O157:H7 and Salmonella spp.): FAM, HEX, ROX, Cy5 and Cy5.5.
- Specio 4.1 (Salmonella spp., L. monocytogenes, E. coli and S. aureus): FAM only, because differentiation is done by the melting curve, not by color.
The practical question is whether your instrument reads those channels. In a single-target kit with two channels, such as Ampliora 1.1, the data sheet or the package insert must indicate which channel corresponds to the microorganism and which to a control.
Compatible thermal cyclers
This field states on which instruments performance was verified. A multiplex kit depends on the instrument having an optical channel for each fluorophore: Ampliora 3.9 uses five, so the thermal cycler must read all five, and its data sheet declares the Bio-Rad CFX96.
The TAAG catalog has a case that shows why this field matters. Ampliora 3.5 (V-SF59) declares the Agilent AriaMx and Bio-Rad CFX96, as well as Open qPCR, with FAM, HEX, ROX and Cy5. Ampliora 3.5R (V-SF74) targets the same three microorganisms, but declares the QuantStudio 5 Real-Time PCR System and uses FAM, VIC, ROX, TAMRA and Cy5, with a PCR time of 110 minutes instead of 100. It is the same panel adapted to another platform: they are not interchangeable.
If the laboratory's thermal cycler is not listed on the data sheet, that use is not validated and the evidence must be requested from the manufacturer.
PCR time and incubation time: not the same thing
PCR time is the figure that says least about when the result arrives. The Ampliora 1.1 data sheet lists a PCR time of 100 minutes and, in the incubation field, a dash. It is not that there is no enrichment: the PCR kit simply does not include it. Enrichment is on the data sheet of the associated medium, Augmentis 14 Universal Gram negative, which declares 24 ± 2 hours of incubation. Augmentis 91 BPW medium, associated with Ampliora 3.9, declares 24 ± 2 hours in general and at least 8 hours for cocoa samples.
To estimate time to result you have to add up at least three data sheets: the medium, the extraction kit and the PCR kit, plus hands-on time. PCR times in the catalog range from 80 minutes (Ampliora 4.7 Low-pH Microorganisms) to 120 minutes (the Specio kits, for example); Elevia 1.1, which works on RNA, declares 95 minutes, the same as Ampliora 2.3.
Sensitivity: what "1 CFU/25 g" means
The sensitivity of a qualitative kit is almost always expressed relative to a test portion: Ampliora 1.1, Ampliora 3.5 and Specio 4.1 declare 1 CFU/25 g in foods; Ampliora 3.9 and Elevia 1.1, 1 CFU/375 g. For surfaces, the data sheets of Ampliora 1.1, Ampliora 3.5, Specio 4.1 and Elevia 1.1 express sensitivity per sample: 1 CFU/sample. It does not mean that the PCR detects one cell in the tube, but that the complete protocol, enrichment included, detects it starting from that contamination in that portion. That is why 375 g and 25 g are not comparable without looking at each one's enrichment protocol.
Some data sheets separate two sensitivities. Ampliora 4.3 Yeast declares 1x10⁰ CFU/mL in enriched samples and 1x10³ cells/mL in direct samples: these are two different figures that should not be confused. And the WaterScan panels express their sensitivity as a range, from 5x10⁰ to 5x10¹ CFU/mL.
LOD, LOD50 and RLOD
In a qualitative method, detection at very low levels is probabilistic: with few cells per portion, some portions contain them and others do not. ISO 16140-2 compares the level of detection of the alternative method with that of the reference method through the RLOD, which it defines as the ratio between the two LODs. The study uses artificially contaminated samples at a minimum of three levels; the low level should ideally sit at the theoretical detection level, 0.7 CFU per portion, and must give between 25 % and 75 % positives with the reference method. The acceptability limit is 1.5 in paired studies and 2.5 in unpaired studies; an LOD lower than that of the reference method is always accepted. AOAC, for its part, expresses performance as probability of detection (POD): the proportion of positive results for a given matrix and analyte level. LOD50 and its uncertainty are covered in limit of detection and LOD50.
Inclusivity and exclusivity: specificity in numbers
A data sheet may say "detection of Salmonella spp.", but that phrase rests on a specific study:
- Inclusivity. The ability of the method to detect the target microorganism across a wide range of strains. ISO 16140-2 requires at least 50 pure cultures of the target and, for Salmonella methods, at least 100 cultures of different serotypes. Each strain is inoculated at a level of 10 to 100 times the minimum detection level of the method and processed with the complete protocol, including enrichment.
- Exclusivity. The lack of interference from relevant non-target strains. ISO 16140-2 requires at least 30 pure non-target cultures. AOAC defines it as a study with potentially cross-reactive non-target strains that must not be detected.
The standard also requires strains to be chosen according to the method's measurement principle. When reading a data sheet, the useful question is with how many strains and serotypes detection was demonstrated and against which non-targets. That information is usually in the validation report, not in the summary data sheet, and it is reasonable to ask for it.
Validated matrices: the field that defines the real scope
The matrices on TAAG data sheets are specific. Ampliora 1.1 declares raw chicken thigh, frozen beef burger, sausage (longanicilla), mortadella and surface. Ampliora 3.9 declares cabbage, lettuce, spinach, a leafy greens mix (equal parts lettuce, cabbage and spinach) and raw ground beef (80 % lean). Specio 4.1 lists a long series of prepared foods, meats, dairy products, pasteurized liquid egg and living and inert surfaces, and its AOAC matrices are stainless steel surfaces, pasteurized liquid milk with 2 % fat and leafy vegetables (lettuce). Elevia 1.1 declares hazelnut-type filled chocolate, milk-type filled chocolate, chocolate bar with peanuts, hazelnut and cocoa cream, whole muscle meat, ground beef (90 % lean), animal feed pellets, pasteurized liquid eggs and stainless steel surfaces.
This matters for two reasons. The first is inhibition: a review by the German Federal Institute for Risk Assessment (Schrader and colleagues, 2012) describes how inhibitory substances present in the sample can reduce PCR sensitivity or produce false negatives, and how some are specific to certain sample types, which calls for matrix-specific extraction protocols. The second is background flora and the condition of the cells: ISO 16140-2 requires a validation to include products with high and low background flora, different process stresses and raw products.
The standard also sets the scope: if a method is claimed for a "broad range of foods", at least five categories must be studied, with at least three types per category. Regulation (EC) 2073/2005 accepts alternative methods validated against the reference method and, if they are proprietary, certified by a third party according to ISO 16140 or other similar internationally accepted protocols. If your product is not among the matrices, the data sheet does not say the kit fails: it says nobody has demonstrated it yet. Verification in your own laboratory, according to ISO 16140-3, is explained in validation and verification of methods.
Storage and shelf life
All PCR kits in the TAAG catalog are stored between -25 °C and -15 °C. The declared shelf life is 9 months for most of them (for example, Ampliora 1.1, Ampliora 3.9 and Specio 4.1) and 6 months for the Elevia kits. Media have their own conditions: Augmentis 14 is stored between 2 °C and 25 °C with a 6-month shelf life, and Nucleia 2 Tez-Q Plus between 2 °C and 25 °C with 12 months. The shortest shelf life in the set determines purchasing.
Certifications and data sheet version
Ampliora 3.9 declares "Certified by AOAC by extension of License Number 032501" and Specio 4.1, "Certified by AOAC by extension of License Number 072101". The Elevia 1.1 data sheet published on taag.bio adds "Certified by AOAC by extension of License Number 072601", with a PTM scope of 62 % cocoa chocolate, cocoa liquor, skimmed milk powder and peanuts (375 g each) and 100 cm² stainless steel surfaces, without the "zero" protocol. The certified scope and the list of validated matrices are different fields and need not match. The word "extension" indicates that the scope of an existing license was broadened, and each certification covers the matrices and targets evaluated in that study, as explained in what AOAC-PTM certification is. In an audit, the license number, the certified matrices and the laboratory's actual matrix must be cross-checked.
Data sheets also have a revision number and date. The Ampliora 1.1 data sheet, for example, is at revision 4, dated 7 October 2025. It is good practice to file the current version together with the verification records.
Which TAAG kits and technology appear in this article
Ampliora 1.1 Salmonella spp. (V-SF97) and Ampliora 3.9 (V-SF68) are real-time PCR assays with fluorescent probes designed with Mila; Specio 4.1 (V-SF95) uses an intercalating fluorophore with a melting curve interpreted with KAi; Elevia 1.1 Salmonella spp. (V-PAT04) is RT-qPCR on RNA with AiGOR technology. In all four cases, the data sheet states that the TxA software interprets the curves automatically. If the laboratory's matrix or instrument does not match those declared, verification is required before using the result.
Conclusion
The technical data sheet of a PCR kit describes the conditions under which the method was demonstrated: with which medium and which extraction, on which instrument and with which channels, for which portion and which matrices, and with which inclusivity, exclusivity and detection level study. Reading it well means separating PCR time from total time, understanding that sensitivity refers to a portion and a complete protocol, and cross-checking the matrices and the certification against what the laboratory actually tests. When something does not match, the data sheet does not say the kit fails, but that it needs to be verified before making decisions with its result.
About TAAG
Review with TAAG the technical data sheet of Ampliora 1.1 Salmonella spp., Ampliora 3.9 or Specio 4.1 against your matrix and your thermal cycler.
Frequently asked questions
Is the PCR time on the data sheet the total time to result?
No. The PCR time covers only the run on the thermal cycler. To the total time you have to add enrichment, which appears on the culture medium's data sheet (for example, 24 ± 2 hours for Augmentis 14 Universal Gram negative), extraction and hands-on time.
What does a sensitivity of 1 CFU/25 g mean?
That the complete protocol, with its enrichment, detects the microorganism starting from that contamination in a 25 g portion. It does not mean that the PCR detects a single cell in the tube, and detection at such low levels is probabilistic, which is why standards evaluate it with LOD50, RLOD or POD.
What are the inclusivity and exclusivity of a kit?
Inclusivity is the ability to detect the target microorganism across many different strains; ISO 16140-2 requires at least 50 strains, and 100 serotypes in the case of Salmonella. Exclusivity means the method does not respond to relevant non-target strains; the standard requires at least 30.
Can I use a kit on a matrix that is not listed on the data sheet?
You can, but that use is not backed by the manufacturer's validation. Before using the result to make decisions, the laboratory must verify the method on its matrix, for example according to ISO 16140-3, because PCR inhibition and background flora change from one food to another.
Does a kit work on any real-time thermal cycler?
Not necessarily. The instrument must read the fluorescence channels the kit uses and be among the instruments on which performance was verified. In the TAAG catalog, Ampliora 3.5 and Ampliora 3.5R target the same microorganisms but declare different platforms and fluorophores.