MYCO-Direct 1.7

DNA based Identification of  Mycobacterium tuberculosis complex (MTUB) ,and other Mycobacteria (MOT)

• Sensitivity
  Short amplicon length, suited for formalin-fixed, paraffin-embedded (FFPE) tissue sections
  MTUB complex 126 bp (IS 6110)
  MOT ~ 225 bp (ITS)

• Specificity
  3 genus specific capture probes, 2 MTUB specific capture probes
  13 capture probes for Mycobacteria other than M. tuberculosis (MOT)

 

• Fast Protocol
  optimized 45 minutes protocol including analysis (post amplification)

 

•  Simple & Robust
  non-fluorescent detection for easy read-out, minimal lab instrumentation required

 

Product Information

Assay Principle

Biotinylated PCR products are generated from sample material containing extracted bacterial gDNA. The labeled amplicons are hybridized to genus/species specific capture probes immobilized on the LCD-Chip surface. Following a short wash routine, comprising high stringency, visualization of bound amplicons is mediated by an enzyme-substrate cascade. Each LCD-Chip contains eight identical micro arrays separated in small reaction chambers. Each array can be addressed individually, allowing the parallel analysis of eight samples on one chip.

 

Target Region & Sensitivity

The targets amplified by the supplied PCR* primer mixes are well conserved sequences and the probes spotted on the array have been thoroughly evaluated for their specificity. Two primer sets for PCR amplification are provided. The „Genus“ mix which targets the rRNA spacer region (ITS) of Mycobacteria, producing an amplicon of 225 bp and the „MTUB“ mix for amplification of a short fragment from the IS6110 element. Due to the short fragment length of both amplicons, the assay is very well suited for the detection of Mycobacteria in formalin-fixed, paraffinembedded (FFPE) tissue sections.

 

Probe Specificity

Fig.1:

Parallel analysis of eight samples on one LCD-Chip. The array field spacing is compatible with standard multi pipettes.

Fig.2 :

Detection of Mycobacteria from formalin-fixed, paraffinembedded (FFPE) tissue sections. A) M. tuberculosis (1 + 2: genus probes 1 and 2; 3: ITS probe for MTUB complex; 4: IS6110 probe) B) M. intracellulare (1 + 2: genus probes 1 and 3; 3 + 4: ITS probes for M. avium complex (MAI)

* The Polymerase Chain Reaction (PCR) process is covered by U.S. patents owned by Hoffmann-La Roche, Inc. and patents owned by F. Hoffmann-La Roche Ltd. No license under these patents to use the PCR process is conveyed expressly or by implication to the purchaser by the purchase of this product.

 

Software

 

SlideReader Vers. 7.01

• Fully automated image analysis
• comprehensive data reports in HTML format
• fast & intuitive navigation
• multiple user accounts

 

Instrumentation

CHIP-Scanner PF2700

• Transmission light scanner for LCD-Arrays,
• 10 µm resolution
• 16 bit grey scale TIFF images

CHIP Spin FVL2400

• Bench Top mini centrifuge
• 2400 rpm, constant
• Adaptor for LCD-Arrays
• incl. 3 rotors