SEROLOGY

NARINDER SINGH
 LIST OF TESTS
1. HCV
2. HBsAG
3. HIV
4. GHB
5. WIDAL TEST
6. RF DETECTION
7. ASO
8. VDRL
9. S.TYPHI IgM
10. MALARIA P.F./PAN
 HCV
Hepatitis c virus tri dot is used for the detection of
antibodies to C virus in serum of the patient
Hcv is spherical, ssrna virus causes acute and chronic
hepatitis.
Procedure :
3 drops of buffer solution
1 drop of serum of patient
5 drops buffer
2 drops protien a conjugate
5 drops of buffer
PRINCIPLE
Immobilized hcv antigens are present on the membrane.as the
patient’s serum pass through the membrane, HCV Antibodies
if present in serum bind to antigens . In washing step unbound
Antigens are removed. Protein-A conjugate is added which
binds to HCV antibodies to give pink colour dot at test region
T1 or T2.
Non reactive resultl: Only one dot at “C”
Reactive results : a} 2 dots at C &T1
b}2 dots at C & T2
c} 3 dots at C , T1 & T2
Invalid : if no dot appear
 HBsAg
Hepacard is an immunoassay test for the detection of hepatitis B
surface antigens in serum.
This helps to diagnose infections due to hepatitis B virus
Procedure includes 2 drops ofserum in the Sample well and read
results after 15 minutes.
Non reactive : one pink line at “C” only .

Reactive results :pink coloured line at T & C region

Invalid : when control line does not appear

 HIV
HIV TRI DOT test is immunoassay for the detection of HIV-1
&HIV-2 Antibodies in serum of the patient .it is screening test
for the hiv.
Principle:Immobilized hcv antigens are present on the
membrane.as the patient’s serum pass through the
membrane, HCV Antibodies if present in serum bind to
antigens . In washing step unbound Antigens are removed.
Protein-A conjugate is added which binds to HCV antibodies to
give pink colour dot at test region T1 or T2.
Procedure :
3 drops of buffer solution
1 drop of serum of patient
5 drops buffer
2 drops protien a conjugate
5 drops of buffer
Non reactive resultl: Only one dot at “C”
Reactive results : a} 2 dots at C &T1(reactive to hiv 1)
b}2 dots at C & T2(reactive to hiv 2)
c} 3 dots at C , T1 & T2 (reactive to both )
Invalid : if no dot appear
 GHB(HbA1c)
• The bio-rad D-10 Hemoglobin is
used for th determination of HbA1c
in whole blood using ion-exchange
high performance liquid
chromatography (hplc).
• Separation of hba1c is done.
Haemoglobins are separated on
basis of their ionic interactions.
• used for the diagnosis of diabetes
mellitus includes deficiency of
insulin and hyperglycemia
• The level of hba1c is proportional
to average glucose concenteration
Referance range
Non-diabetic :<5.7
pre-diabeic : 5.7-6.4
diabetics : >or = 6.5
MPG
Mean plasma glucose is estimated
from the hba1c value
Mpg=28.7(hba1c%)-46.7
for Eg;
Mpg = 28.7 (5.8)-46.7
Mpg = 119.76 mg/dL
 Widal test
• For detemination of Antibodioes to S.typhi(O&H antigen) and
S.paratyphi(AH &BH antigen) in serum
• Based on the principle of direct agglutination.when patient’s
serum (containing antibodies to s.typhi &s.paratyphi) is mixed
with respective antigens (O,H,AH&BH). Agglutination
indicates the presence of antibodies
• Slide test: includes 4 circles O,H,AH&BH
• Place one drop of serum in each circles
• Add one drop each O,H,AH &BH in each circles
• Mix and rock the slide gntly for 1 minute and observe
agglutination
 BIOLOGICAL REFERANCE
RANGE: <1:80 TITRE

POSITIVE RESULT:
CARPET FORMATION
O (DISLODGE)

H
NEGATIV RESULT:
AH BUTTON
FORMATION
BH

1:80 1:160 1:320 1:640 1:1280 C

 RF DETECTION
It is a rapid agglutination
procedure for the detection
of rheumatoid factors.
Principle:antigen coated with
human gamma globuin
,agglutinates in the presence
of rheumatoid factors in
patient serum
Place 1 drop of serum in circle
and add one drop of RF latex RF SENSITIVITY : 10 IU/Ml
in circle Positive result : presence of agglutination
Mix and rock the slide for 2 Whch means conc. Of RF in sample is equal
minutes Or greater than 10 IU/mL
Negative result: no agglitination
Rf conc. Less han 10 IU/mL
 Other tests
• VDRL TEST :syphilis is STD .after
the infectionhost forms
treponemal antibodies and when
serum is reacted with trepolipin
reagent , flocculation reaction is
produced
• ASO TEST: Aso latex reagent is a
agglutination procedure for the
detection of antistreptolysin-
o.streptolysin o is toxic produced
by beta haemolytic streptococcus
bacteria.
Aso sensitivity : 200 IU/mL
 s.typhi IgM:
• enterocheck:used to detect typhoid
fever caused by S.typhi & S.paratyphi.
It is immunoassay for the detection of
IgM antibodies to S.typhi in serum

Malaria antigen test

p.f./pan cassete is used to detect malaria
Which contains membrane strip which is
precoated with one monoclonal
antibody (specific to p.falciparum) and
polyclonal antibody for (plasmodium
species) as two separate lines across a
test strip.
 Rf dilution 100 ul normal saline

100uL serum
100 uL 100 uL 100 uL
100 uL

1:2 1:4 1:8 1:16 1:32

For eg; Antibody titre is 1:4
Then, conc of RF = titre * sensitivity
conc.=4* 10 IU/mL = 40 IU/mL
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