/ College of Engineering Biomedical engineering

Evening study

2019/2020

Report in the Biomaterials Engineering course

By:

Mustafa98

second stage

Supervision :
Introduction crucial Currently,the spread of the viral transmission
become fast so that the prevention of local transmission
the third Zoonotic human coronavirus requires apoint-of care test (POCT),which shows quick
(CoV)of the century emerged in outcome within 20 minutes . COVID-19Ab test is an in
December 2019,with a cluster of vitro diagnostic medical device that helps you to
patients connected to wuhan , Hubei diagnose Noval Coronavirus infections quickly and
province, China. this virus ,the newly accurately by measuring the lgG or lgM antibody for the
identified corona virus 2019 nCOV, 2019-nCoV.
could cause risky pneumonia so that
prevention and control of the infection The Aim of this way to test materiel
has become highly required .the 2019- methods
nCOV is a member of the
Betacoronavirus Genus ,that also crating way to detect Co_19 virus the current test
includes severe Acute Respiratory method is by using PCR called polymerase chain
syndrome corona virus (SARS-COV)and reaction
middle east Respiratory syndrome
coconavirus (MERS-CoV).Since it is
identified that symptoms become
rapidly severe without a proper
treatment after onset of illness, early
diagnosis of the virus infection is quite
1
method its very In this case we try to detect covid-19
sensitive and can find using sandwich immunodetection
infection period Even method, fluorescence-labeled
in the early phase of conjugates in dried detection buffer
disease and take binds to antibody in sample forming
hours to obtain, so antibody-antigen complex ,and migrates
the staff doctors have onto nitrocellulose matrix to be capture
waste sever all hours by other immobilized anti human iGg
to find if the patient and anti human IGm on test cartridge
infected or not this ,the more anti bode in the sample foms
wasted time can be the more antigen-antibody complex and
used to treat the leads to stronger intensity of
patient Even its on the fluorescence signal on detector antigen,
early time of Disease which is processed by to show
so the other way is to concentration of anti-Ncov iGg and IGm
use antibodies that in sample respectively
present in the body
that’s creates to fight Diagnosis Antibodies .. antibody’s
the virus .. are protein that bind target
material(microbes .viruses..) by specific
2. Methods and Mateial structure we can generate Antibodies
by injecting the mouse ,rabbit with virus
Principle so that immune system of this Animal
will produce specific antibodies to this
virus according to the physiology . A
single rabbit can produce . A million
antibody’s that we can use selective
way to screen AB candidates by
laboratory Department we can develop
vary of antibodies FIG.1
and by clinical microbiology
Department We can grow the virus and
produce Antibodies to produce test not
only for covid_19 but also for wide
range of Disease and infection (HIV,HCV,
anti HBs ,PCT CEA ect Fig.1 Antibody Structure

Latral flow Immunoassay

?{LFDs}.
1
 not test line to find if presence of
Simple stricture to diagnostic Device use
Analyte (Antigen ).Many type of LFDS
to know if presence or absence of
are found But the commen one is
Analyte Such as Bacteria .pathogens
sandwich assays .It is Properties it is
.ect... in sample { plasma .Serum .Blood
that the posetive test Represent by
} LFDs it is Structure contain control Line
present of analyte in the sample . How
to confirm if the strip work properly or
does the flow work)..Lateral flow assays
have a wide array of applications and
can test a variety of samples like urine,
blood, saliva, sweat, serum, and other
fluids. They are currently used by clinical
laboratories, hospitals, and physicians
for quick and accurate tests for specific
target molecules and gene expression.
Other uses for lateral flow assays are
food and environmental safety and
veterinary medicine for chemicals such
as diseases and toxins.Lateral flow tests
are also commonly used for disease
identification such as ebola, but the
most common lateral flow test is the
home pregnancy test.and currently used
to test covid-19 (FIG 2)

FIG .2 A rapid lateral flow immunoassay

1
 Obtain Antibodies
Rabbits are well-known for munity
sorting immune Response Against
foreign Amplified and Rabbit polyclonal
AB are among the lirs Established and
most widely used For Reserch tool in
immunology interms of Affinity and
Specificity Rabbits produce superior AB
against many antigen ,the majority
antibodies is immunoglobulin Igg within
developing lymphocytes and which is
leads to develop the secondary AB IGm
.Manufacturing process Monoclonal
AB.. as show in FIG.3
FIG.3 Recombinant Rabbit Monoclonal
Antibodies
3 Test Principle and Procedure .
Covid-19 test can Detect targets using
fluorescence immunochrona to graphic
based on lateral flow assay that involve
disposable test strip .100 ML of the
sample was add to the sample pad
internal control, both dried reagent and
containing dried fluorescence -labeled
detect or antibody then the sample
move on to the test strip by the capillary
the target antigen or antibody was
present in the sample they will react
with
the fluorescence -Conjugted Antibody
or Antigen to form Antigen Antibody
complex on the nitrocellulose
Membrane ,Fluorescence intensities of
the target marker and Control Material
were detectedusing fluorescence
Reader .Line of the Cartridge indicate
adequate sample and capillary
movement the Result appear via
calibration Statistic Based on the values
of Sevially diluted Standard material
with other Device Like (PCR ).The
calibration curve as used in Analytic
chemistry also known as standard curve
used to Determined the concentration
of substance in an unknown Sample
Comparing with known Result as We
compare with (PCR) ..By using
equation of cal Curve the equation will
be General in form of(Y =MX+b) Where
M is slop (b) is constant So the
concentration that will be optioned is
(Y= MX+b)
4.Automated Fluorescent
Immunoassay System
The cartridge(LFDs) contains a
detection buffer including the dried
reagent and monoclonal IgY as an
monoclonal IgY are labelled with europium
 chelate[Eu(III)] , When mixed with the cut-off index
sample, the detector binds to target
(COVID-19) in the sample, forming
antigen-antibody complexes.
When the complexes migrate onto the
nitrocellulose matrix, the other capture
(antigen or antibodies) immobilized on
the “Test line”that containing IGm,IGG
that are specific to Covid-19 forms a
sandwich complex. The scanner [FIG.4
A] quantifies the analytes by measuring
the fluorescence intensity on the test
strip induced by a laser by using light
source to stimulate the conjugated
fluorescence material that bind to the [FIG.4 A] The scanner
secondary antibody , so the target
material will emit light when stimulated
and this light can be picked up by a
detector . The fluorescence detector
[FIG.4B] has a fixed absorption
wavelength of 333 nm and an emission
wavelength of 613 nm, Fluorescence of
the target material its intensity is
proportional to the concentration of the
target molecules in the samples. The
result of the samples is given as
“positive,” “negative,” or [FIG.4 B] Optic detector .
“indeterminate” in the form of a relative

Result result numerically in terms of cut-off

index (COI) value as follows
According to the boditech Company as
they make Covid-19 test so they ahve
been developed great way to test this
virus ,test result qualitatively as
‘Negative’ or ‘Indeterminate’ or
‘Positive’ while also displaying test
 COVID-19 Ab IgM *COI Negative or
Indeterminate or Positive
COVID-19 Ab IgG *COI Negative
or Indeterminate or Positive

2019-nCOV RT-PCR assay

Test result is negative if COI is < 0.9, Posi Nega Tot
indeterminate if COI is 0.9-1.1 and Positive
tive
46
tive
0
al
4
6
positive if COI is >1.1-200. AFIA
Negative 0 145 14
S COVID-
5
19 Ab
Indetermin 2 5 7
ate
Total 48 150 19
1. COVID-19 Ab test result is ‘Negative’ 8
- Positive Percent Agreement: 95.8%
for IgG as well as for IgM, the patient
- Negative Percent Agreement: 96.7%
may not be infected with anti-SARS-
CoV-2 but follow-up testing with a
molecular diagnostic test should be
considered to rule out infection in these
individuals.
2. If the test result is ‘Indeterminate’
for IgG and/or for IgM, the patient may
be infected with anti-SARS-CoV-2 and
hence there is need to retest another
sample from the patient.
3. If the test result is ‘Positive’ for IgG
and/or for IgM, the patient is most likely
to be infected with anti-SARS-CoV-2.

Clinical performance evaluation

COVID-19 Ab has demonstrated
following clinical performance results
when clinical samples collected from
various asymptomatic individuals and
patients suspected of COVID-19 disease
were tested with COVID-19 Ab test with
analyzer and confirmed by testing with
Allplex™ 2019-nCoV Assay
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