Gold Np’s in Diagnosis
Invitro Diagnosis
Name : Veera CSR, Chittepu
Out line
• Nanodiagnosis is defined as application of
nanotechnology to research in Diagnosis.
• Gold particles usage in Diagnosis, which are
at commercial level, approved by FDA.
• Current research in this area.
• Companies who made this technology at Point
of care level, used in daily life
F
Functionalization of Au-NPs with biomolecules
for clinical diagnostics.
& antibodies for signal enhancement in immunoassays
& carbohydrate functionalization to study specific molecular
Interactions (Carbohydrate functionalized nanoparticles
(“glyconanoparticles” have the advantage of increasing the specific
interactionsbetween glycans and lectins for biosensing applications)
Under development
& surface functionalization with ligands that can be
tailored for specific protein binding or direct binding of peptides and
proteins to the Au-nanoparticle surface
&The specific interaction between biological pairs has also
been widely used,
e.g. biotin–streptavidin and Ni-NTA–histidine tail .
Current on going Research on use of
Au Np’s
• Fraunhofer IBMT have developed a new
integrated biosensor system.= Accurate Diagnosis
of prostate cancer using Optoacoustic detection
of biologically functionalized gold Nanoparticles.
The treatment relies on optoacoustic imaging of
gold nanoparticles.
• Use of ELISA technology for liver cancer diagnosis
used gold nanoparticles ringed by a charged
polymer. coating and an X-ray scatter imaging
technique to spot tumor-like masses as small as 5
millimeters earlier it was 5 cm
Researchers at Northwestern
University
• an ultrasensitive, automated immunoassay that is
capable of detecting serum PSA at concentrations as
low as 330 fg/ml—approximately 300 times more
sensitive than commercially available immunoassays.
Use of Bio-barcode PSA assay. To detect PSA in serial
serum samples from 18 patients who had undergone
radical prostatectomy for prostate cancer. PSA was
detected in 102 (86%) of the 118 serum samples tested
using the bio-barcode assay, compared with just 30
(25%) when commercial assays were used.
• UGA researchers use gold nanoparticles to diagnose flu
in minutes
Commercial companies by
WGC
• Church and Dwight - First response pregnancy
kit.
• BBInternational andMerck - detection of food
prothegens
• Merck single path assay for presence of
salmona , within 20 minutes And duopath
assay can be used for salmonella, E.Coli and
Campylobacter.
• NanoSphere - Verigene detecting specific
biomolecule targets with gold nanoparticles
NanoSphere
Nanosphere develops, manufactures and markets
an advanced molecular diagnostic platform,
the Verigene System, which enables simple, cost-
effective, and highly sensitive nucleic acid (DNA
and RNA) and protein testing on a single
platform.
• The direct detection of single or multiple nucleic
acid targets with single-base pair resolution has
diagnostic applications in several clinical areas,
including:
• The detection of DNA or RNA targets using nanoparticle probe technology entails:
• Automated nucleic acid extraction and PCR amplification (for some assays) from a clinical sample
on the Verigene® Processor SP
• Automated transfer of eluted nucleic acids or amplicons into a single-use, proprietary Test
Cartridge. The DNA is then cut via a proprietary sonication process, into 300-500 base-pair
fragments in order to facilitate hybridization in subsequent steps.
• In the initial hybridization step, the cut genomic DNA (target nucleic acid) is simultaneously
hybridized to:
– Single-stranded DNA capture oligonucleotides arrayed in replicate on a solid support (an array), and
– Sequence-specific mediator oligonucleotides that detect single-copy DNA regions of each target of interest.
– A washing step then follows that removes unhybridized gold nanoparticle probes.
• Silver signal amplification is then performed on the gold nanoparticle probes that are hybridized to
captured DNA targets of interest.
• There then follows a washing step to remove unreacted signal amplification reagents.
• Qualitative analysis of results (slide reading) can now be performed on the Verigene®Reader.
Gram-positive Blood cultured
Gram Negative Bacteria
FDA not cleared still.
Cardiac Tests
warfarin test
n=238) clinical
samples in
comparison to bi-
directional DNA
sequence analysis
of the same
samples. The
percent agreement
between the two
methods was
100%
Human Genetic Test
warferin metabolism
(n=238) clinical
samples in
comparison to bi-
directional DNA
sequence analysis of
the same samples.
The percent
agreement between
the two methods was
100%
Verigene System Light Cycler Instrument
where nanoparticles are
illuminated using a fixedwavelength
light source
No Chemical amplification of chemical
Signal Single-image sensor
sample size : 25 µl
Analysis Time : 2 minutes
DNA amplification Via PCR
Optical detection of stimulated
fluorescence.
Sample Size : 10-20µl
Analysis Time : Detection occurs at defined
intervals during PCR cycle and can be
viewed in real-time
Some Patent : METHOD FOR DETECTING THE PRESENCE OF A TARGET ANALYTE IN
A TEST SPOT - and also patent on Nano particle imaging system and method.
Gram positive blood
culture Nucleic acid test
FIRST RESPONSE GOLD DIGITAL PREGNANCY TEST
CHURCH & DWIGHT CO., INC.
Regulation Number 862.1155
• early detection of pregnancy by the detection of hCG, a placental
hormone in urine. The device detects the presence of hCG in the
urine of a pregnant woman by way of a series of immunochemical
reactions on a chromatographic strip contained within a plastic
housing, which is integral with the digital component that reads and
displays the result of the immunochemical reactions on the Display
Screen of the device in a manner similar to the predicate device.
• The test is indicated for use from four days before the expected
period, that is, five days before the missed period. The intended use is
Invitro Diagnostic device intended for detection of hormone presence.
• sensitivity of 18 mIU/ml, accuracy is only 58 percent if test is meant 5 days
before, for 100 percent accuracy, we have to wait for biological cycle to
reach maximum concentration of harmone.
FIRST RESPONSE EARLY RESULT PREGNANCY TEST (OTC)
CHURCH & DWIGHT CO., INC.
Regulation Number862.1155
• The test result is read in the housing window after the elapse Of 3
minutes. Two pink lines indicate hCG has been detected (pregnant);
one pink line indicate not detected.
• DrawBack : The test may detect the pregnancy hormone (hCG), in some
cases, as early as 6 days before the missed period (5 days before the
expected periodicates no hCG has been detected (not pregnant).
Note : Some pregnant women will not be able to detect hCG in their urine 6
days before the missed period. If you test negative before your missed
period, but think you may still be pregnant, you should re-test again a few
days after your missed period.
this test detects very low levels of hCG, there is a small chance that this
test will give positive results even if you are not pregnant. Chances of
this are greater for women nearing age 40 and older.
• Church and Dwight used Gold Nanoparticles to detect hcG hormone, they also use strepdavin, and
the only device which can detect low level so f hcG hormone.
•
ABBOTT , Abott advisor one step pregnancy test kit. here in place of gold coated colloidal partciles,
colloidal are coated with anti alpha hcG antibody (related to mouse)hormones.
WH Accu Test the sensitivity of the test is 100%, it takes five minutes to read test.
• Recently, some researchers used WBC to test pregnancy(Joseph P. Habboushe, MD Department of
Emergency Medicine, New York Hospital–Queens of Cornell University, 56-45 Main St, Queens, NY
11355, USA)
• Regarding patent information, they author has got 3 patents, Authors are Nazareth, US and Albert
Nazareth, Mercerville, NJ US. Here they are using gold nanoparticles whose sizes are 40-47 nm
and 60-75 nm .
Single Path E.Coli O157
Gold labelled Immuno sorbent Assay
• Used in food analysis laboratories, qualitative
detection of E. Coli from a variety of foods
• Immunochromato Graphic rapid Test based on
gold labelled anti bodies. Here They get gold
labelled particle from BBinternational.
• The test device has Circular sample port, oval
Shaped test T, and control window C
The sample is applied to the chromotography paper via sample
port. The sample is observed through the pad to the reaction
zone containing Gold labelled antibodies specific to Ecoli. Any
E. coli antigen present complexes with gold antibody and
migrates through the port until it encounters a binding zone
in test area T. The Binding zone T contains another anti E coli
antibody, which immobilize any E. Coli complex present. Due
to gold labelling distinct red line is formed. The rest of the
sample migrates to second binding reagent zone within the
control Zone C and forms second distinct line
Sample
Care must be take when working with
samples, culture and media
• Detection limit – 25 gram of sample
• Interferences : no interference of Singlepath E.Coli with
food ingredients.
• Sensitivity & Specificity & Efficiency - >99%(according
to AOAC trials)
• False negative rate and False positive rate < 1%.
• Problem : No line appears after 20 minutes of the test
period., Measure : re run sample again
• AOAC = association of analytical communities
• Duo Path Assay is used to detect salmonella, E.Coli and
Campylobacter
Flow diagram showing food brone pathogens How
previous method is useful
B BioAssayWorks
• BioAssay Works (Ijamsville, Maryland)
develops lateral-flow, point-of-care and point-
of-use diagnostic tests, with emphasis on
assays requiring high sensitivity.
• KIM-1, a whole new dimension in kidney
damage detection. Available in ELISA or rapid
Rena-strip test format.
BioAssayWorks
H-Rena-strip™ and
R-Rena-strip™
Lateral-Flow Tests
Fast–test results in just 15 minutes,
Reliable–specificity 96%–sensitivity 99%,
Easy-to-use–no special training
necessary, Semi-quantitative–results can
be correlated with color chart,
Quantitative results–with reader (sold
separately)
H-Rena-E™ and R-Rena-E™ ELISA Tests
• Fast–test results in just 3 hours•
Reliable–specificity 100%–sensitivity
100%
• Quantitative results
C-FLAT® Counter-Flow Lateral Amplified
Test: C-FLAT is a patented lateral-flow
immunodiagnostic device
References
• World Gold Counsil
• FDA
• Corresponding company websites.
• For patents, usa patent database, and
company website.
Thank you and Happy X- Mas
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