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Recall 3
Recall 3
E. nana:
E. histolytica:
3.Maximum interval which a recipient sample maybe used for crossmatching if the patient has
been recently transfused, has been pregnant? 3 days
9. CPT blood was drawn @ 10 Am, pooled @ 11:30 am. Patient has xray @ 2 pm, what will you
do with the product?
- transfuse the blood before xray
10. Enzymes:
Destroy = M N S Duffy
Enhance = Rh Lewis I Kidd
- LE
- RF
- IM
- Infectious Hepatitis
- Leprosy
- Malaria
- Pregnancy
- Aging process
- Pneumococcal pneumonia
In this sample notice the speckled ANA is the dominant pattern in the interphase cells
(a) and some speckling in the area outside of the chromosomal area of the mitotics (b).
Also notice the smooth staining of the chromosomal area of the metaphase mitotic
cells (c). This represents the presence of a homogeneous ANA pattern.
a. Peripheral (rim) – the central protein of the nucleus is only slightly stained or not
stained at all , but nuclear margins fluoresce strongly and appear to extend into
the cytoplasm
- i.e. anti – DNA
- associated with SLE in the active stage of the dis. and in Sjogren’s dis.
d. Nucleolar - a few round, smooth nucleoli that vary in size will fluoresce when
examined with UV.
- i.e. anti- nucleolar
- present in 50% with Scleroderma, Sjogren’s syndrome, SLE
-
-
In order to understand the ANA test (antinuclear antibody test), it is first important to
understand different types of antibodies.
Antibodies are proteins, produced by white blood cells, which normally circulate in the blood to defend against
foreign invaders such as bacteria, viruses, and toxins.
Autoantibodies, instead of acting against foreign invaders, attack the body's own cells. This is an abnormality.
Antinuclear antibodies are a unique group of autoantibodies that have the ability to attack structures in the
nucleus of cells. The nucleus of a cell contains genetic material referred to as DNA (deoxyribonucleic acid).
An ANA test (antinuclear antibody test) can be performed on a patient's blood sample as
part of the diagnostic process for certain autoimmune diseases.
To perform the ANA test, sometimes called FANA (fluorescent antinuclear antibody test), a
blood sample is drawn from the patient and sent to the lab for testing. Serum from the
patient's blood specimen is added to microscope slides which have commercially prepared
cells on the slide surface. If the patient's serum contains antinuclear antibodies, they bind to
the cells (specifically the nuclei of the cells) on the slide.
A second antibody, commercially tagged with a fluorescent dye, is added to the mix of
patient's serum and commercially prepared cells on the slide. The second (fluorescent)
antibody attaches to the serum antibodies and cells which have bound together. When the
slide is viewed under an ultraviolet microscope, antinuclear antibodies appear as fluorescent
cells.
ANA Titer
A titer is determined by repeating the positive test with serial dilutions until the test yields a
negative result. The last dilution which yields a positive result (fluorescence observed under
the microscope) is the titer which gets reported. Here is an example:
1:10 positive
1:20 positive
1:40 positive
1:80 positive
1:160 positive (reported titer)
1:320 negative
Positive or negative
If positive, a titer is determined and reported
The pattern of fluorescence is reported
ANA titers and patterns can vary between laboratory testing sites, perhaps because of
variation in methodology used. These are the commonly recognized patterns:
Homogeneous - total nuclear fluorescence due to antibody directed against nucleoprotein. Common in systemic
lupus erthematosus (SLE).
Peripheral - fluorescence occurs at edges of nucleus in a shaggy appearance. Anti-DNA antibodies cause this
pattern. Also common in SLE.
Speckled - results from antibody directed against different nuclear antigens.
Nucleolar - results from antibody directed against a specific RNA configuration of the nucleolus or antibody
specific for proteins necessary for maturation of nucleolar RNA. Seen in patients with systemic sclerosis
(scleroderma).
ANA test results are just one factor considered when a diagnosis is being formulated. A
patient's clinical symptoms and other diagnostic tests must also be considered by the
doctor. Medical history is also significant because some prescription drugs can cause "drug-
induced antinuclear antibodies".
Statistically-speaking, the incidence of positive ANA test results (in percent per condition)
is:
Subsets of the ANA tests are sometimes used to determine the specific autoimmune
disease. For this purpose, a doctor may order anti-dsDNA, anti-Sm, Sjogren's syndrome
antigens(SSA, SSB), Scl-70 antibodies, anti-centromere, anti-histone, and anti-RN.
The ANA test is complex, but the results (positive or negative, titer, pattern) and possible
subset test results can give physicians valuable diagnostic information.
Several different serum tests are used to detect autoimmunity. These are conditions where the
immune system acts directly against the bodys own tissues. One test, the ANA, or anti-nuclear
antibody test, detects antibodies that are directed against various components of the nucleus of
the cell. These include antibodies that have been formed against double-stranded or single-
stranded DNA (two ways in which the cells DNA can be found in the serum after being released
from old and dying cells). Other components of the nucleus such as histones are also released
from old cells and can also become targets of the immune response. When they appear they
may be markers for excess or inappropriate immune responses directed against ones own
tissues. Physicians in our group test for various autoantibodies in order to characterize patients
as those who might have a tendency for autoimmune responses. Those who test positive have
been found to have a higher risk for recurrent pregnancy loss and are more likely to benefit from
therapeutic interventions (see diagram).
Consequences
A blood test determines the presence of antibodies to polynucleotides, histones and DNA. This
process involves running 27 different tests on a sample of blood.
The presence of antibodies is also tested for by doing the ANA test. This is a less sensitive test
but one that many doctors have already done on their patients before we ever see them.
The test is reported as a titer and a pattern. Any titer above 1:40 is significant. The titers can get
into the thousands such as 1:2,500. This simply means that the test is positive when the blood
serum is diluted many times.
In women with autoimmune diseases these antibodies cause inflammation in joints and organs.
In women with no autoimmune diseases but a positive antibody, the antibody causes
inflammation around the embryo at the time of implantation or in the placenta after implantation.
This inflammation is exactly the same as occurs if you get a splinter under your fingernail. The
tissue around the splinter gets hot, red and swollen and it happens quickly.
15. Read about MIC for susceptibility test to report in microbiology
16. Epstein Barr Virus, How to detect? Know the procedure.
- infectious mononucleosis
- test to detect:
a) Mono spot
b) CBC
c) EBV serology
- can help detect if an individual has an infection due to EBV, and if they are prone
to future infections due to dormant virus.
- VCA-IgM
VA-IgG Tests ---------- help to identify current infection
EA-D
EBNA Test -------------- help to dx future infection due to an existing dormant virus.
d) throat culture
e) Liver profile
-
I M V C:
E. coli + + - -
KES - - + +
Shigella + + + +
Salmonella - + - -
Edwardsiella + + - -
- Sensitization
= 1st step in agglutination
= physical attachments of antibody molecules to antigens on the RBC membranes
- Lattice Formation
= establishments of cross – links between sensitized particles and antibodies
resulting in aggregation (clumping), is a much slower process thant the sensitization
phase
30. Dilution:
1st tube 2nd tube 3rd tube 4th tube 5th tube 6th tube
0.1 Serum 0.5 serum
0.9 diluent 0.5 diluent
32. Green top tube , blood is collected and refrigerated for 3 hours. Should you not accept?
My answer: plasma should be separated before refrigeration
38. automaterd method for measuring Chloride which generates silver ions in the reaction.
- cystic fibrosis
- Coulometry
42. Which of the following methods is MOST reliable for determining the appropriate
dosage of Rh immune globulin to give to an identified Rh immune globulin candidate
after delivery?
Flow cytometry is the most reliable method of those listed. It is a quantitative method,
whereas Keihauer-Betke and the rosette test are very subjective tests.
- Drugs
48. Machines set @ 150 wavelenght, then wavelength @ 0 when used. What is the
problem?
Hemochromatosis
Iron deficiency anemia
Anemia of chronic diseases
Thalassemia
- multiple myeloma
- presence of Bence jones protein in the urine
- monoclonal gammopathy
Bone marrow cytology in a dog with multiple myeloma. There are large numbers of plasma cells (*) in the
aspirate, some of which are binucleate. Hematopoietic cells (both myeloid [M] and erythroid [E]) are found
in normal numbers and maturation sequence.
I.Labs: Findings
A. Serum Protein Electrophoresis and Urine Protein electrophoresis for
Monoclonal Peak
1. M Protein in either serum or urine: 97% of patients
2. Serum M Protein by electophoresis (82%) or immunofixation
(93%)
3. Urine M Protein by electrophoresis: 75%
B. Chemistry panel with Serum Calcium
1. Hypercalcemia
a. Serum Calcium >11 mg/dl (present in 13% of patients)
Renal Insufficiency
. Serum Creatinine >2 mg/dl (present in 23% of patients)
C. Complete Blood Count with platelets
Normochromic Normocytic Anemia
. Hemoglobin <12 grams/dl (present in 65% of patients)
D. Bone Marrow Aspiration and biopsy
E. Peripheral Smear
Myeloma Cells
Rouleaux of Red Blood Cells
F. Erythrocyte Sedimentation Rate (ESR)
Increased >50 mm/hour in most cases (except bence-jones Myeloma)
G. Serum Viscosity
H. Urinalysis
Bence-Jones Protein
Serum:
-Immunoelectrophoresis
-Immunofixation
-Quantitation of immunoglobulins by radial immunodiffusion or nephelometry
- Screening for croglobulins
- Determination of serum viscosity of IgM, IgA, or IgG , or signs and symptoms suggestive of
Hyperviscosity
Urine:
- Screening of urine for increased protein, e.g. sulfosalicylic acid
- Total protein assay of a 24 hour urine specimen
- Urinary protein electrophorsis
- Urinary immunoelectrophoresis
- Immunofixation
DIC
CLL
Smudge cells
Big Platelet
66. Ferritin :