MOLgen

Please pay attention to the revision of the document that must be the same as the revision reported in the box label.
In case of discrepancy please contact our Customer Care e-mail: info@adaltis.net.

* Other document related to the product available at Documentation Centre and it is accessible for Adaltis distributors/partners after registration only.

MOLgen MTBC/NTM Real Time PCR Kit is designed for the qualitative simultaneous detection of DNA of MTB Complex (MTBC) and non-tuberculous mycobacteria (NTM) of clinical interest (Mycobacterium abscessus, Mycobacterium massiliense, Mycobacterium parascrofulaceum, Mycobacterium kansasii, Mycobacterium tuberculosis, Mycobacterium avium complex, Mycobacterium intracellulare, Mycobacterium gordonae, Mycobacterium senegalense, Mycobacterium smegmatis, Mycobacterium szulgai, Mycobacterium marinum, Mycobacterium malmoense, Mycobacterium xenopi, Mycobacterium fortuitum) by Real Time Polymerase Chain Reaction (PCR) from DNA extracted from decontaminated human respiratory specimens such as sputum and bronchoalveolar lavage fluid (BALF).

Tubercolosis (TB) is caused by Mycobacterium tuberculosis complex (MTBC) and it mainly affects the lungs. Mycobacterium tuberculosis complex (MTBC) may cause Tubercolosis in human and other animal range hosts: human hosts (infection caused by M. tuberculosis, M. africanum, M. canettii), bovine hosts (infection caused by M. bovis), caprine hosts (infection caused by M. caprae), rodent hosts (infection caused by M. microti), seal hosts (infection caused by M. pinnipedi).
TB is spread through droplets and aerosol in the air when people with lung TB cough, sneeze or spit. A person needs to inhale only a few germs to become infected. Every year, 10 million people fall ill with tuberculosis (TB). Despite being a preventable and curable disease, 1.5 million people die from TB each year – making it the main cause of pathogen agent death in the world.
Besides, TB is the leading cause of death for people with HIV and also a major contributor to antimicrobial resistance.

MTBC/NTM Real Time PCR Kit is intended for the detection of the strains of MTBC and NTM present in the different geographical areas using real-time polymerase chain reaction (Real Time-PCR) method with fluorescence detection of amplified product.
During the DNA polymerase synthesis of the complementary strand, the probe is cleaved from the 5’-end due to the 5’-3’ esonuclease activity of Taq DNA polymerase used, the quencher and the dye become separated thus increasing the fluorescence signal due to accumulation of the reaction product. The detected fluorescence intensity depends on the initial quantity of pathogen’s DNA template in the sample.
The primer and probe are designed for amplification of two conserved regions (IS6110 and MBP64) of MTB Complex.
Primers and probe for amplification of non-tuberculous mycobacteria (NTM) of clinical interest are designed on a conserved region (Intergenic Spacer Region), which specifically recognizes atypical mycobacterial strains.
The detection of amplified targets fragment is performed on channel FAM (IS6110 and MBP64), HEX (ISR/ITS).
The use of human beta actina as Internal Control (IC) on CY5.5 channel ensure the monitoring correct amplification and cellularity of the samples.
The results of PCR analysis are taken into account in complex diagnostics of disease. The kit is validated for use with: AMPLIlab (Adaltis).

The kit contains reagents for 25 rxns (code METBNT25)
or 100 rxns (code METBNT100).

1. Molgen MTBC/NTM Master Mix

Ready to use reagent.

Contains reagents for amplification.

2. Molgen MTBC/NTM Probe Mix

Ready to use reagent.

Contains primers and probes which are premixed to the working concentrations to identifying MTBC and NTM.

3. Molgen MTBC/NTM Positive Control

Ready to use control.

Contains plasmids with IS6110, MBP64, (ISR/ITS), sequences and a plasmid with sequences of b-actin as Internal Endogenous Control (IC).

4. Molgen MTBC/NTM Negative Control

Ready to use control.

Negative Control should be included in every run as it indicates that reagents have not been contaminated.

Nucleic Acid extraction kit intended for isolation of DNA (MOLgen Universal Extraction Kit (Adaltis) Ref. ME188830).

· Decontaminating and fluidifying reagent.

· Real-time PCR cycler.

· Biosafety Cabinet.

· Refrigerator.

· Microcentrifuge with rotor for 1,5 mL reaction tubes.

· 1.5 mL microcentrifuge tube for extraction.

· Plate shaker.

· Calibrated adjustable pipettes.

· Disposable pipette tips with aerosol filter.

· Strip Tubes and Caps (0.2 mL) or PCR Tubes (0.2 mL) or 96 well plate with optical caps or films.

· Disposable medical non-sterile powder-free gloves.

· Biohazard waste container.
NOTE: Automated extraction with MOLgen Universal Extraction Kit (Adaltis) Ref. ME188830 has been validated with the ELAB (Extraction) and EXTRALAB (Automated Extraction and PCR setup) available from Adaltis S.r.l.. Extraction systems are installed with the extraction protocols preloaded.

WARNING ANDPRECAUTIONS

         Strict compliance with the instructions for use is required for optimal results. Any deviation from these instructions may affect results.

• Use of this product is limited to personnel specifically instructed and trained in the techniques of real time PCR and in vitro diagnostic procedures.
• Use of this product is limited to personnel specifically instructed and trained.
• Correct Personal Protection Equipment (PPE) must be worn when handling specimens and reagents in accordance with local regulations for biosafety procedures and handling pathogens.
• Carefully read the Safety Data Sheet (SDS) and instructions for use before product use.
• Do not mix the components from kits of different lots.
• Avoid more than 3 thawing- freezing cycles.
• Treat all specimens as potentially infective using safe laboratory procedure. Refer to Interim Laboratory Biosafety Guidance for Handling and Processing Specimens Associated with 2019-nCoV.
• To prevent cross-contamination it is recommended to have dedicated work areas for sample preparation ( DNA extraction) and PCR Amplification (setting up PCR reaction and thermocyclers).
• To conduct real-time amplification reaction with PCR products detection, use only disposable tips with aerosol filters.
• Dispose unused reagents and waste in accordance with country, federal, state and local regulations.
• Do not use the kit after the expiration date.

STORAGE AND TRANSPORTATION

· Transport the kit at -15°C…-25°С.

· Store the kit in the manufacturer’s packaging at -15°C…-25°С for the entire shelf life.

· Do not freeze the Kit more than 3 times