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
2Myconostica 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
3Fungal 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.
4Myconosticas 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.
5Technology
- 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
6The 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
-
7MycXtra 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)?
8DNA amplification and detection
- Low level multiplex
- Real time PCR
- High level multiplex
- DNA biochip/Array
9Molecular Beacons
10High Quality DNA
Multiplex Real Time PCR with Molecular Beacons
11 MycAssay
12Microarrays
- 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
13Microarrays
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
14Microarrays
- System comprises customised low density
microarrays and an inexpensive, dedicated reader
for colourimetric detection
- discrimination depends on differential
hybridization of target and probes
15MycArray (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
16MycArray (Candida) hybridized with both C
parapsilosis and control DNAs from
asymmetric PCR
17Product 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 19Backup
20MycXtra Fungal DNA Extraction kit
21PCR process