1. PRECISION DEVICE FOR THE RAPID DIAGNOSTIC OF MICROVILLUS INCLUSION DISEASE
Theodore J Hall, Biomedical Engineer
Dr. Michael Caplan Ph.D. , School of Biological Health Systems and Engineering
David Carpentieri, M.D. , Mitchell Shub, M.D. , Phoenix Children’s Hospital
-Microvillus Inclusion Disease (MVID) is a
rare genetic disease which leads to a non-
functioning brush border in the small
intestine.
-Due to the 1977 CàT single nucleotide
mutation, this disease un-proportionally
affects people of the Navajo nation.
-Current diagnosis methods have patients
waiting over 4 weeks for results, a critical
window for patient survivability. With
implementation of the Tentacle Probe (TP),
diagnostic cost and time is reduced to $100
and 24 hours.
Background
Equations
Legend
pr is the probability of reaction; v is the average velocity
R is the sum of the radii of the two molecules; P is the probe concentration
T is the target concentration; kf is the association rate constant
[1] Erickson, Robert P., et al. American Journal of Medical Genetics. Part
A: Navajo Microvillous Inclusion Disease is due to a Mutation in MYO5B.
146A Vol. Wiley-Liss, 12/15/2008. Web. 22 Nov. 2015.
[2] Satterfield, B.C., J.A.a. West, and M.R. Caplan. “Tentacle Probes:
Eliminating False Postives without Sacrificing Sensitivity.” Nucleic Acids
Research(2007). Print..
[3] Christine Beeson, Jeffrey C. Weiss, Robert D. Ligensky. Microvillus
Inclusion Disease- A Deadly Masquerador.
[2]
Works Cited[2]
[2]
Future Work
Methods
-Diagnostic device will follow the steps: 1) DNA
acquisition via blood sample 2) DNA purification 3)
rt-PCR analysis of DNA with TP 3) output result of
patient diagnosis
-Development of wild type probe has the ability to
predict the likelihood of passing down the mutation to
further generations
Sequence of the TP:
(5’
-‐
[BHQ1]
-‐
ccgCAAGCGTTGAGCTTGcgg
-‐
[orange
560]
-‐
[linker
9]
-‐
GACATAGTGAGGTGTCGTGGCA
-‐
3’)
The TP combined with DNA containing the target mutation was run through a melting curve.
As temperature increased the probe and DNA melted, allowing for a binding event to occur.
The binding event demonstrates that the probe will bind to the target mutation
-Tentacle Probe (TP) is used in a rt-PCR
machine. When the probe binds to the target
mutation it will fluoresce; as shown in the melt
curve.
-The mutation is detected by the levels of
fluorescence. The resulting exponential
fluorescence curve serves as the diagnostic
determinant.
-Since an rt-PCR machine is used to amplify the
DNA and the TP is used to detect the mutation, a
rapid and accurate diagnostic result can be
obtained
Results
![PRECISION DEVICE FOR THE RAPID DIAGNOSTIC OF MICROVILLUS INCLUSION DISEASE
Theodore J Hall, Biomedical Engineer
Dr. Michael Caplan Ph.D. , School of Biological Health Systems and Engineering
David Carpentieri, M.D. , Mitchell Shub, M.D. , Phoenix Children’s Hospital
-Microvillus Inclusion Disease (MVID) is a
rare genetic disease which leads to a non-
functioning brush border in the small
intestine.
-Due to the 1977 CàT single nucleotide
mutation, this disease un-proportionally
affects people of the Navajo nation.
-Current diagnosis methods have patients
waiting over 4 weeks for results, a critical
window for patient survivability. With
implementation of the Tentacle Probe (TP),
diagnostic cost and time is reduced to $100
and 24 hours.
Background
Equations
Legend
pr is the probability of reaction; v is the average velocity
R is the sum of the radii of the two molecules; P is the probe concentration
T is the target concentration; kf is the association rate constant
[1] Erickson, Robert P., et al. American Journal of Medical Genetics. Part
A: Navajo Microvillous Inclusion Disease is due to a Mutation in MYO5B.
146A Vol. Wiley-Liss, 12/15/2008. Web. 22 Nov. 2015.
[2] Satterfield, B.C., J.A.a. West, and M.R. Caplan. “Tentacle Probes:
Eliminating False Postives without Sacrificing Sensitivity.” Nucleic Acids
Research(2007). Print..
[3] Christine Beeson, Jeffrey C. Weiss, Robert D. Ligensky. Microvillus
Inclusion Disease- A Deadly Masquerador.
[2]
Works Cited[2]
[2]
Future Work
Methods
-Diagnostic device will follow the steps: 1) DNA
acquisition via blood sample 2) DNA purification 3)
rt-PCR analysis of DNA with TP 3) output result of
patient diagnosis
-Development of wild type probe has the ability to
predict the likelihood of passing down the mutation to
further generations
Sequence of the TP:
(5’
-‐
[BHQ1]
-‐
ccgCAAGCGTTGAGCTTGcgg
-‐
[orange
560]
-‐
[linker
9]
-‐
GACATAGTGAGGTGTCGTGGCA
-‐
3’)
The TP combined with DNA containing the target mutation was run through a melting curve.
As temperature increased the probe and DNA melted, allowing for a binding event to occur.
The binding event demonstrates that the probe will bind to the target mutation
-Tentacle Probe (TP) is used in a rt-PCR
machine. When the probe binds to the target
mutation it will fluoresce; as shown in the melt
curve.
-The mutation is detected by the levels of
fluorescence. The resulting exponential
fluorescence curve serves as the diagnostic
determinant.
-Since an rt-PCR machine is used to amplify the
DNA and the TP is used to detect the mutation, a
rapid and accurate diagnostic result can be
obtained
Results](https://image.slidesharecdn.com/c4e9569f-6dfe-4bae-b842-99f3bb6ebad1-160424224917/85/Hall-Theodore-1-320.jpg)
