Explanation of DNA Sequencing for ENT Speciality.

1. Next Generation-Sequencing of
Bacteria, Fungal ,Viral

2. Dr. Caroline Fife
Dr. John Steinberg Dr. Chris Attinger
Dr. James McGuire
Wound Care ENT
Dr. David Kennedy : Chairman Penn
Dr. BJ Ferguson U of Pittsburgh
Dr. Martin Citardi Chairman U ofTexas
Dr. Donald Lanza
Dr.Takashima Director SinusCenter Baylor
Dr .WinstonVaughn Stanford SinusCenter
House Ear Institute Los Angeles CA
ENT Dept. Georgetown Medical School
Dr Patrick Antonelli – Chief Otolaryngology UF

4. “In this case, the
results of Next
Generation
Sequencing
contributed directly
to a dramatic effect
on the patient’s
care, resulting
ultimately in a
favorable
outcome”.

6. Pathogenius DNA Molecular Diagnostics.
25,000 DNA codes on file. Indifferent to time, media and transport issues.

7. Otitis Media Biofilm
Sinus Cavity Biofilm

8. The Role of Bacterial Biofilms and the Pathophysiology of
Chronic Rhinosinusitis
Shaun J. Kilty, MD, and MartinY. Desrosiers, MD
“The concept of bacteria living in the organized
communities that compose bacterial biofilms is a
radical departure from the classic notion of bacterial
infection and bacterial behavior.”
“When in the form of a biofilm,
the causative bacteria are difficult to culture and are
largely resistant to current antimicrobial therapy.”

9. Planktonic Bacteria
Single Cell
Bio Film
“Community of Bacteria”
 Cause of 80% of Infections
 Cause of ChronicWounds
 Will not grow in traditional culture
 Biofilm cells express a radically different
phenotype than planktonic bacteria
Only Diagnostic tool is Molecular
 Cause of Acute Infection
 Will grow in traditional Culture

10. The majority of the bacterial cells in most microbial
communities grow in the biofilm phenotype,
These cooperative processes are facilitated by physical
connections between the individual cells like nanowires
(Gorby et al. 2006
Goodman et al. 2011 ), and any attempt to rip one of
these resolutely interactive cells from its designated bed in
the biofilm would result in a group of “dazed and
confused” bacterial cells that would be unable to grow in
any medium.
BioFilm Research

19. Biocides vs. Biofilm

20. DNA Analysis vs.Traditional Cultures
51 Chronic wounds- Parallel Samples
16 S DNA Sequencing Traditional Culture
• 46 /51 Staph Identified
• 32 / 51 Pseud Identified
• 28/51 Staph Identified
• 8 / 51 Pseud Identified
DNA Sequencing OutperformedTraditional Cultures
DNA Sequencing identified 145 Genera – Cultures 14 !

22. “The impact of proper
specimen management on
patient care is enormous”
“Specimen selection and
collection are the
responsibility of the
medical staff and not the
laboratory”

23. American Society for Microbiology Standard:
Swab transport, RoomTemperature, 2 hours
Current Lab: Swab transport, unknown
temp, end of the day.
High probability of error from the start.

24. Diagnostic Services – What we deliver
Two molecular microbial diagnostic tests are currently used at PathoGenius®. We take
your patient's swab, washing, or tissue sample; extract the microbial DNA; and report
the microbial analysis to you.
Level 1: PCR 24-Hour Rapid Screening
Screen 8 Common ENT Bacteria and 1 Fungal
Provide Quantitative Bacterial Load
Rapid turn around time of approximately 24 hours from the time of sample
receipt.
Pathogens Identified in Level 1
Staphylococcus (including MRSA)
Streptococcus pyogenes (GAS)
Haemophilus influenza
Moraxella Catarrhalis
Pseudomonas
Streptococcus pneumonia
Streptococcus agalactiae
Candida albicans

25. “I need Sensitivity for antibiotics"
• “Biofilm-growing microorganisms are
significantly more tolerant to antibiotics [6] and
corresponding breakpoints have not been
established [34].The S-I-R results can therefore
not be used to predict therapeutic success in the
case of biofilm infections and offer no guide to
clinicians for treating such infections.”
ESCMID Guidelines for the diagnosis and treatment of biofilm infections

26. Dead Bacteria DNA
Bacterial Biofilms and Chronic Rhinosinusitis Kilty
and Desrosiers
“Most convincing was the demonstration in a chinchilla model
that live bacteria, although nonculturable, could persist in
OME for weeks, whereas DNA strands and DNA from intact
but nonviable bacteria could not exist for more than a day
[21].”
“Given the extremely short half-life of mRNA, its presence was
evidence that viable metabolically active bacterial organisms
were likely present in OME.”

27. Benefits Pathogenius DNA MolecularTesting
• Fast Identification of Biofilms and their composition in
patients referred to you—Accuracy and Speed.
• GOLD STANDARD of Microbial Diagnostics
• Medicare Covers theTest and pay $190
• Less Expensive than a Culture

28. COST AND COVERAGE OF
MOLECULAR DIAGNOSTICSTRADITIONAL CULTURE
• MEDICARE PAYS
$250
DNA SEQUENCING
• MEDICARE PAYS
$ 285

29. CUSTOMIZED FOR EACH
PATEINT TOPICAL RX
LIPOGEL BASED
DOSED 3X WEEK
CONTAINS ANTIBIOFLIM AGENTS
ANTIBIOTICS
ANTIFUNGALS

30. DNA Diagnostic “Must be expensive “
How much will it cost?
Private Office
If your patients are a Medicare patient, 100% of the
cost of your testing will be paid by Medicare.
Medicare reimburses $190 for our Level 1 and Level 2
If your patients have private insurance, 50% of
Commercial Plans pay for the test.
No sample will ever be denied by Patho Genius
No Patient will ever go to collection

31. Benefits Pathogenius DNA MolecularTesting
• Fast Identification of Biofilms and their composition in
patients referred to you—Accuracy and Speed.
• Gives you the most comprehensive bacterial and fungal
and viral commercially available
• Eliminates errors and misdiagnosis caused by sample
handling: Proper Media,Time, temperature and
transport as required by the American Society of
Microbiology.
• Proven to improve healing times and clinical outcomes

32. Questions?

33. Lab Requisition