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Detection and Identification of Infectious Disease Workshop

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Aspergillus. Pneumocystis. Rare fungi, such as Zygomycetes ... unsuccessful because Aspergillus and other fungal DNA was detected. in digestion controls' ... – PowerPoint PPT presentation

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Title: Detection and Identification of Infectious Disease Workshop


1
  • Detection and Identification of Infectious
    Disease Workshop
  • 24th March 2009
  • Adrian Moody
  • Director of Research and Development

2
Myconostica Group
  • The Myconostica Group is developing medical
    diagnostics for harmful fungal diseases. Founded
    in 2006 it is based in
  • Charleston SC
  • Facility being constructed for US manufacturing
    and distribution-current total construction
    contract gt300k-finish date April 2009
  • Manchester, UK
  • Research and Development
  • Assembly

3
Fungal infections a major medical problem
  • Bloodstream fungi
  • Candida
  • Cryptococcus
  • Histoplasma
  • Rare fungi, such as Fusarium
  • Lung fungal infections
  • Aspergillus
  • Pneumocystis
  • Rare fungi, such as Zygomycetes
  • Fungal disease is a growing and an increasingly
    important area of medicine
  • Significant cause of mortality in
    immunosuppressed patients
  • Emergence of resistant infections to antifungal
    agents
  • Inadequate methods to diagnose primary infections
    and monitoring response of infecting organism to
    therapy.

4
Myconosticas product pipeline addresses key
unmet needs
  • SENSITIVITY of diagnosis
  • SPEED of diagnosis
  • SPECIFICITY of diagnosis

Reducing the time period from specimen collection
until species identification and anti-microbial
susceptibility, is critical to improving outcome
of patients with invasive infections.
5
Technology
  • Technology agnostic
  • Our vision To be recognised as a world leader in
    clinically accepted diagnostics for life
    threatening fungal infections
  • Current focus molecular biology
  • DNA extraction
  • DNA amplification and detection

6
The Need for a Dedicated DNA Extraction System
  • Attempts to use the xxxxx extraction method on
    BAL fluid were
  • unsuccessful because Aspergillus and other fungal
    DNA was detected
  • in digestion controls
  • Fredricks et al. (2005) J. Clin. Micro. 43
    5122-5128
  • 2,800 specimens extracted, 1 in 5 a negative
    control
  • 3.3 contaminated during extraction, 4.7
    contaminated during
  • amplification
  • Loeffler et al. (1999). J. Clin Micro.
    371200-1202
  • A dedicated extraction system provides known
    characteristics
  • Suits user requirements
  • Yields and sensitivity
  • QC tested for contamination
  • Removal of interfering substances

7
MycXtra DNA extraction kit
  • Extracts and purifies fungal DNA from clinical
    respiratory samples (BAL and sputum) for
    subsequent PCR analyses
  • Features
  • 10 tests/kit
  • CE marked
  • 18 month shelf life
  • Minimal equipment required Vortex Genie 2
    Adaptor plate
  • (Myconostica product code 080-015)?

8
DNA amplification and detection
  • Low level multiplex
  • Real time PCR
  • High level multiplex
  • DNA biochip/Array

9
Molecular Beacons
10
High Quality DNA
Multiplex Real Time PCR with Molecular Beacons
11
MycAssay
12
Microarrays
  • Why use microarrays for identifying fungal
    pathogens?
  • DNA microarray technology allows the simultaneous
    detection of many targets
  • clinical need for species identification to
    predict antifungal treatment efficacy
  • identification of resistance markers gives more
    detailed information

13
Microarrays


MycXtra
streptavidin-enzyme conjugate Substrate conjugate
buffer enzyme buffer

Rapid detection of Candida spp
PCR primers enzyme dNTPs including
biotin-16-dUTP internal control
Candida spp
14
Microarrays
  • System comprises customised low density
    microarrays and an inexpensive, dedicated reader
    for colourimetric detection
  • discrimination depends on differential
    hybridization of target and probes

15
MycArray (Candida)
  • 14 species of Candida plus 4 relevant
    non-Candida species
  • Initial indications in vitro and positive blood
    culture
  • FDA pre-IDE meeting in April
  • CE marking and US launch 2H09

16
MycArray (Candida) hybridized with both C
parapsilosis and control DNAs from
asymmetric PCR
17
Product Availability
  • Myconostica has a strong pipeline of products for
    launch into global markets over the next 3-5
    years.

Currently on market in US for Research
As part of MycAssayTM (Aspergillus)
18
  • Thank you and Questions

19
Backup
20
MycXtra Fungal DNA Extraction kit
21
PCR process
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