Mycobacteria Identification

MYCO-Direct 1.7 LCD-Array Kit

Reliable and sensitive identification of members of the Mycobacterium tuberculosis complex (MTBC), Mycobacterium avium-intracellulare complex (MAC) and 10 mycobacteria other than tuberculosis (MOTT). Three aditional capture probes (Mycobacteria I-III) directed against conserved motifs within the ITS region of mycobacteria can furthermore indicate the presence of DNA from other mycobacteria without providing species information. Due to small amplicon sizes and the combination of two robust PCR systems, the assay is well suited for the analysis of formalin-fixed, paraffin-embedded (FFPE) tissue samples.

The specific detection range can be broadened to more mycobacterial species (e.g. M. fortuitum, M. gastri, M. leprae and others) by simply hybridising the PCR products generated with primer mix A (ITS) to MYCO-Plus 2.0 arrays without the requirement for an additional PCR. (see below for more details).

PCR coupled DNA macroarray (reverse hybridisation). Biotinylated PCR products are generated from nucleic acid extracts of sample material containing mycobacterial DNA. The labeled amplicons are hybridized to 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 which can be addressed individually.

The assay is based on two PCR reactions which are combined prior to hybridization into one array field. Primer mix A directed against the internal transcribed spacer (ITS) region of mycobacteria and primer mix B directed against the insertion element IS6110 exclusively found in members of the MTBC. Both PCRs generate amplicons of less than 350 bp length (Mix A, ITS: ~200-300 bp, Mix B, IS6110: 127 bp) and allow for the detection of 50 genome equivalents (GE) per reaction for MOTT (ITS) and 1 GE for MTBC (IS6110).

Brochure DNA Array Mycobacteria MTBC

LCD Array Kit

MYCO-Direct 1.7

Identification of Mycobacterium tuberculosis complex (MTBC) & 11 mycobacteria other than tuberculosis (MOTT)

Order codes:

32 Tests   [ B-500-04 ]

96 Tests   [ B-500-12 ]

  Brochure   MSDS

Spotting pattern DNA Array MYCO-Direct 1.7

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Analysis Screen Shots

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Related Literature

  • Comparison of 14 Molecular Assays for Detection of Mycobacterium tuberculosis Complex in Bronchoalveolar Lavage Fluid (2013). Akkerman O.W., van der Werf T.S., de Boer M., de Beer J.L., Rahim Z., Rossen J.W.A., van Soolingen D., Kerstjens H.A.M. and van der Zanden A.G.M. J. Clin. Microbiol.,51(11),3505-3511   LINK:   PDF at jcm.asm.org
  • Identification of HPV Types and Mycobacterium Tuberculosis Complex in Historical Long-Term Preserved Formalin Fixed Tissues in Different Human Organs (2017). Huehns M., Erbersdobler A., Obliers A. and Roepenack P. PLoS One, 12(1):e0170353   LINK:   PDF at plos.org

MYCO-Plus 2.0 LCD-Array Kit

Identification of 9 additional mycobacteria on the species level by using PCR products of primer mix A (ITS) from the MYCO-Direct 1.7 kit for hybridisations against MYCO-Plus 2.0 arrays. No extra PCR required. The MYCO-Plus LCD-Array Kits are available as supplemental products only. The kits contain all detections chemicals required for hybridisation and staining, but no primer mixes.

See MYCO-Direct 1.7

See MYCO-Direct 1.7 (PCR with primer mix A directed against mycobacterial ITS region)

LCD Array Kit

MYCO-Plus 2.0

Identification of 9 additional Mycobacteria

Order codes:

32 Tests   [ B-650-04 ]

96 Tests   [ B-650-12 ]

Accessory Product to MYCO-Direct 1.7

No additional PCR required

Spotting pattern DNA Array MYCO-Plus 2.0

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Analysis Screen Shots

Screenshot Analysis LCD-Array MYCO Plus

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Periodontitis

BAC-Dent 2.4 LCD Array kit

Progressive destructive periodontal diseases are caused by different factors and influences. Several bacterial species and their metabolic products are triggering factors for periodontal and peri-implantary tissue loss. Among those a group of highly pathogenic bacteria are considered to be in strict correlation with some forms of periodontitis. Infections involving species like Aggregatobacter actinomycetemcomitans and Porphyromonas gingivalis require a specific antibiotic treatment. Detailed knowledge about the species composition of individual subgingival biofilms can contribute to an optimised therapy procedure.

The BAC-Dent 2.4 LCD-Array Kit is an in vitro diagnostic product intended for the parallel detection of 11 bacterial species associated with human periodontitis. By analysing bacterial DNA extracted from subgingival plaques, the kit provides a specific, qualitative assessment of the presence of eleven marker microorganisms in the sample material.

PCR coupled DNA macroarray. Biotinylated PCR products are generated from nucleic acid extracts of  subgingival plaque samples. The labeled amplicons are hybridized to 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 which can be addressed individually.

The assay is based on a triplex PCR directed against three regions of the bacterial 16S rRNA gene generating fragment lenghts of approx. 85 bp, 300 bp and 350 bp (species dependent). Using the recommended PCR protocol with 38 PCR cycle repetitions, ~ 100 target copies (20 for A. actinomycetemcomitans) will reliably be detected. See the detailed instructions for use for more information about the assay sensitivity.

Brochure DNA Array Periodontitis

LCD Array Kit

BAC-Dent  2.4

Identification of 11 Bacteria associated with Periodontitis

Order codes:

32 Tests   [ B-400-04 ]

96 Tests   [ B-400-12 ]

  Brochure   MSDS

Click pattern to enlarge

Analysis Screen Shots

Click image to enlarge

Related Literature

  • Microbial complexes in subgingival plaque (1998). Socransky S.S., Haffajee A.D., Cugini M.A., Smith C. and Kent jr. R.L. J. Clin. Periodontol., 25(2) 134-144   LINK:  onlinelibrary.wiley.com
  • Introduction to microbial aspects of periodontal biofilm communities, development and treatment (2006). Socransky S.S. and Haffajee A.D.  Periodontology 2000; 42(1), 7–12.   LINK:   onlinelibrary.wiley.com
  • Functional biomarkers for chronic periodontitis and insights into the roles of Prevotella nigrescens and Fusobacterium nucleatum; a metatranscriptome analysis (2015). Szafranski S.P., Deng Z.-L., Tomasch J., Jarek M., Bhuju S., Meisinger C., Kühnisch J., Sztajer H. and Wagner-Döbler I. npj Biofilms and Microbiomes 1, 15017.   LINK:   PDF at nature.com