Добавил:
Sekretar
kiopkiopkiop18@yandex.ru
t.me/Prokururor I Вовсе не секретарь, но почту проверяю
Опубликованный материал нарушает ваши авторские права? Сообщите нам.
Вуз:
Предмет:
Файл:Ординатура / Хирургия / Библиотека им академика М.И. Перельмана / Книга_5512_Библиотеки_им_академика_М_И_Перельмана.pdf
Laborato cedures in Serology
O
Specimen collection and storage
DO NOT USE PLASMA, AS USE SERUM.
Principle
Reagents, supplies and equipment
Note
Note
Caution preservative
Procedure
Caution
1s2 I
negative reaction
Medical Laborato Technology: Volume 2
streptococcal exoenzyme
antigen
Principle
ex
tracellular antigens,
Reagents and supplies
Procedure
Laborato cedures in Serology
J1s3
streptolysin O unit
°
Todd unit
Principle
°
Reagents and Supplies
Streptolysin-O:
Do not freeze.
Laboratory supplies other than those available in the kit:
Procedure
°
=
=
=
°
°
Medical Laboratory Technology: Volume 2
784
additional SerodiagnoSiS KitS
Table 24.4 Preparation of dilution series for the anti-streptolysin-O test
Tube
No.
Volume of inactivated
and diluted serum (mL)
1:10
(A)
1 0.8
2 0.2
3
4
5
6
7
8
9
10
11
12
13
14
−
−
−
−
−
−
−
−
−
−
− −
− −
1:100
(B)
−
− −
1.0
0.8
0.6
0.4
0.3
Volume of
buer (mL)
1:500
(C)
0.2 1 : 12.5 0.5 0.5
0.8 1 : 50 0.5 0.5
−
−
−
−
−
1.0 0.0 1 : 500 0.5 0.5
0.8 0.2 1 : 625 0.5 0.5
0.6 0.4 1 : 833 0.5 0.5
0.4 0.6 1 : 1250 0.5 0.5
0.2 0.8 1 : 2500 0.5 0.5
0.0 1 : 100 0.5 0.5
0.2 1 : 125 0.5 0.5
0.4 1 : 167 0.5 0.5
0.6 1 : 250 0.5 0.5
0.7 1 : 333 0.5 0.5
1.5 control 0.0 0.5
1.0 control 0.5 0.5
Resulting
serum dilution
Volume of
reduced SLO (mL)
Volume of 5%
suspension of RBC
Observation and reporting
The end-point is the last tube (i.e., the highest dilution) showing no haemolysis. Control
tube 13 should show no haemolysis. If there is haemolysis in this tube, the test should be
repeated. Control tube 14 should show complete haemolysis.
Adoption to slide agglutination test for titre determination
• Use serum without inactivation.
• Prepare serum dilution as given in Step 1 and Table 24.4.
• Mix one drop of diluted serum and one drop of antigen-coated latex suspension in
order to determine the immunologic reaction with increasing dilution of serum.
• The highest dilution of serum that shows positive reaction is the titre.
• Follow manufacturer’s direction to report the result in Todd units.
Staphylococcus strains are easily cultured in laboratory that takes 1–2 days. Serodiagnosis
kits are now available that can conrm the species of Staphylococcus aureus. It dierentiates
Staphylococci which possess clumping factor and/or Protein A. Blue latex powder pre-coated with the antibody to Staphylococcus aureus (in short Staph aureus) is mixed with a Staph
culture. If a specic antigen of Staph aureus is present, it will react with the latex-antibody,
producing a noticeable agglutination (which is more easily seen with the blue of the latex).
Another test is Clearview Exact Strep dipstick. It is a rapid test used to detect Strep (strep
throat) antigen directly from throat swabs. It is an immunoassay test, consisting of a strip that
has been pre-coated with anti-Strep antibody plus another Strep antibody that is coloured
with gold. The Strep from the patient’s mouth is taken on a coon swab and the Strep antigen
is then extracted with extraction reagents. The test strip is inserted into this antigen reagent
mixture and it is moved chromatographically up the test strip, aaching to the antibody. This
Laboratory Procedures in Serology
785
infeCtiouS MononuCleoSiS and MonoSPot teSt
antigen–antibody complex then binds with the second Strep antibody (bound to gold), forming a ‘sandwich.’ A coloured ‘sandwich’ of antibody–antigen–antibody–gold is formed on the
test lines, showing up as a pink line.
Serological identification of Staphylococcus and Streptococcus
Supplies
• A Staphylococcus culture on an agar medium plate
• A Streptococcus culture on an agar medium plate
• Difco Staph latex-kit or Staphylo slide kit
• Clearview test for Streptoccus
Serological typing of Staphylococcus aureus
Procedure
1. Mix the latex reagent by shaking; expel any latex from the dropper for complete mixing.
2. Dispense 1 drop of Test Latex (contains Staph antibody) onto one of the circles on the
reaction card and add 1 drop of Control Latex onto another circle.
3. Using a loop, pick up and smear 5 suspect colonies (either a suspect Staph from your
body or a known Staph aureus supplied from the lab) onto the Test Latex-containing
circle and mix this into the Test Latex reagent. Spread to cover the circle; be sure to have
a homogenous bacterial suspension. Clumps will make the reaction dicult to identify,
and possibly give a false + reaction.
4. Repeat Step 3 for Control Latex using the same bacterial culture.
5. Pick up and hand rock the card for up to 20 s and observe for agglutination under
normal lighting conditions.
6. Rock the slide back and forth for no longer than one minute and look for blue agglutination.
Clumping is a positive reaction for the identication of Staphylococcus aureus.
(Note An overhead light will facilitate to observe the clumping).
Clearview test for Streptococcal antigen
The kit contains all the required reagents. Only minimum laboratory equipment are necessary.
Procedure
1. Place the extraction tube in the tube holder. Add 3 drops of extraction reagent 1 to the
extraction tube. Add 3 drops of extraction reagent 2 to the extraction tube.
2. Place the throat swab specimen in the extraction tube. Rotate the swab inside the tube
using a circular motion (you may use the pure culture of Strep provided) to roll against
the side of the extraction tube so that liquid is expressed and reabsorbed from the swab.
3. Let it stand for a minimum 1 min.
4. Gently squeeze the swab rmly against the tube to expel as much liquid as possible
from the swab. Discard the swab.
5. Immerse the test strip into the extraction tube with the arrows pointing towards the
specimen extract. Leave the test strip in the extraction tube. Start the timer.
6. Read results in 5 min. Depending on the number of organisms on the swab, a positive
result may be visible as soon as less than 1 min. However, to conrm a negative result
the complete reaction time of 5 min is required. Do not read results after 10 min.
Infectious mononucleosis, often called “mono”, refers to a group of symptoms usually caused
by the Epstein-Barr virus (EBV). It spreads through saliva and found commonly in newly
married couples and often referred as “kissing disease.” People with mono often have high
fever, swollen lymph glands, and sore throat. Fortunately, most cases of mono are mild and
1s6 I
Medical Laborato Technology: Volume 2
Monospot Test (Heterophil Test)
antibody test.
Supplies
Procedure
negative
positive
patient's serum
latex
ote
ote
Acquired lmmuno-Deficiency Syndrome (AIDS)
human
immunodeciency virus
Laborato cedures in Serology
ote
J1a1
788 I
Reagents and supplies
Procedure
Observation and reporting
Medical Laborato Technology: Volume 2
Viral Hepatitis
jaundice
markers
ELISA for hepa B surface antigen
Laborato cedures in Serology
Reagents and supplies
Procedure
Note
J1s9
Precautions
Dipstick test for hepatis B surface anen
red bands.
190 I
Medical Laborato Technology: Volume 2
Reagents and supplies
Procedure
Malaria
Plasmodium.
Dipstick test for malaa
immunochromatographic assay Plasmodium
falciparum, P. vivax P. malariae P. ovale.
P. falciparum
P. vivax P. lciparum
Reagents and supplies
Procedure
Falciparum