Mycobacterium tuberculosis PCR Kit CE/IVD

1 Intended use:
RealCycler MTBC is an in vitro diagnostic kit of reagents which allows real-time PCR qualitative detection of DNA from Mycobacterium tuberculosis complex in clinical samples.
The system includes an internal control CHIC (Competitive Heterologous Internal Control) to prevent false negatives due to reaction inhibition.

2 Principle of the test
The amplification of Mycobacterium tuberculosis complex is detected in the corresponding channel of FAM fluorophore and the internal control is detected in the corresponding channel of Alx532 or HEX fluorophore.

3 Technical specifications
Sensitivity:
1 copy/μL.

Specificity: Mycobacterium tuberculosis complex (IS6110).

Compatibility with DNA/RNA extraction systems

Validated Instruments
- Magcore (RBC Bioscience)>
- QiaCube (Qiagen)
- Arrow/Liaison IXT (DiaSorin)
- Maxwell 16 (Promega)

Compatible Instruments
RealCycler Monotest and RealCycler Universal kits are compatible with almost all DNA/RNA extraction systems available in the market, without making modifications to the original protocols provided by the manufacturer.
This is due to the CHIC Technology (Competitive Heterologous Internal Control), on which the internal control of amplification system is based. This proprietary technology allows the use of any extraction system without loss of sensitivity, and at the same time it monitors accurately a possible sample inhibition.
Even though the CHIC technology enables the monitoring of the quality of the extraction process, it is necessary to verify that your purification equipment or manual extraction system allows you to obtain DNA and RNA with the proper quality and quantity. It is recommended to use external quality controls (QCMD, NIBSC, etc.) in order to conduct a self-validation of your extraction system.

Compatibility with Real-time PCR instruments

Validated Instruments
- Andana 1216 (Progenie molecular)
- Flash 20 (Coyote Bioscience)
- SLAN-96P (Sansure)
- MIC (Biomolecular Systems)
- QuantStudio 5 (ThermoFisher Scientific)
- CFX96 (Bio-Rad)
- ABI 7500 (Life Techonologies)
- AriaDx (Agilent Technologies)
- Rotor-Gene Q (Qiagen)

Compatible Instruments
RealCycler products are compatible with the most used Real-time PCR instruments. The following grid shows the fluorophores used for our products and their corresponding wavelengths. You can check the compatibility of your equipment with the used fluorophores.

CHIC Technology

CHIC technology (Competitive Heterologous Internal Control) is the internal amplification control system developed by Progenie Molecular that includes RealCycler kits.

This proprietary technology allows the use of any extraction system without loss of sensitivity, and at the same time it monitors accurately a possible sample inhibition. For this reason, RealCycler Monotest and RealCycler Universal kits are compatible with almost all DNA/RNA extraction systems available in the market, without making any changes to the original protocols provided by the manufacturer.

Internal amplification control
The false negatives in the PCR-based techniques for the detection of pathogens occur when some factor prevents the generation of amplification product, regardless the presence of the pathogen in the sample. The usual cause is the presence of PCR inhibitors, that could be components of the clinical sample (hemoglobin or other proteins, mucopolysaccharides, urea, etc.) as well as components artificially added during its handling (heparin, EDTA, proteinase K, powder from latex gloves, etc.). Since it is an inherent feature of the samples or their processing, the inhibitions can not be detected unless specific prevention systems are used.

The internal amplification control (IC) is a system for the detection of false negatives. It is based on the simultaneous detection of a fragment different from that of the pathogen, regardless the presence of the pathogen in the sample and therefore, it is always detected. It is made of a DNA or RNA template, oligonucleotides, and the probe that allows its specific detection. The IC can also be used to detect problems in the DNA extraction or its absence in the amplification mix. Its use also allows the prevention of false positives due to the amplification of unspecific products. In the absence of the pathogen's DNA, the IC consumes the surplus reagent, minimizing the amplification of undesired sequences and hindering the generation of a false positive.

The internal control CHIC is an internal control system based on our proprietary CHIC technology (Competitive Heterologous Internal Control). This technology is based on the use of heterologous DNA artificial constructions (made of combined sequences of human and microorganisms DNA) as template for the detection of the internal control. The main feature of this technology is that the amplification of the internal control finds itself in a competitive disadvantage with the pathogen. In this way it does not affect the sensitivity of the method as a whole and it allows the development of highly sensitive systems. RealCycler products have this internal control system in all its formats: RealCycler Monotest and RealCycler Universal. Additionally, this technology allows the use of almost any extraction system without the loss of sensitivity and, at the same time, it monitors accurately a possible sample inhibition.

CHIC features
- Co-amplified in the reaction mix together with the detected pathogens.
- Competitive disadvantage with the pathogen = it does not decrease the sensitivity of the pathogen detection.
- Independent of the extraction system = almost complete compatibility with all extraction systems.
- It monitors inhibition with high accuracy (contact us for examples).