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Which is the main limitation of standard PCR that is not a limitation of real-time
PCR?
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The main limitation of standard PCR is the inability to quantitate the DNA product
due to reliance on endpoint analysis. The final yield of a product is not always equal
to the estimated exponential growth per cycle. Therefore, quantitation is not possible
with endpoint analysis. Real-time PCR overcame this limitation.
Question Difficulty: Level 6
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PCR can be modified for the amplification of RNA with one additional step prior to
the PCR process - the addition of a retrovirus enzyme called reverse transcriptase. The
reverse transcriptase is used to create a copy of DNA using the original RNA
specimen. This method is used for many viruses, such as HIV, that have an RNA
genome. RT-PCR is extremely advantageous for many reasons. First, one can use a
blood sample of less than 1 mL which makes it available to test and monitor
neonates.Second, RT-PCR can detect infection in individuals before antibodies
appear. In addition to HIV, RT-PCR is the primary mode for the detection and
monitoring of hepatitis C.RT-PCR is also used for identifying Mycobacterium
tuberculosis and cytomegalovirus (CMV). RT-PCR is commonly combined with real
time PCR.
Fc
Fab
heavy chains
light chains
Hinge regions
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Hybridization
Extension
Annealing
Denaturation
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Anti-mitochondrial antibody
Immunoglobulin quantitation
Screen for Sm and RNP antibodies
Anti-DNA antibody using C. lucillae
Mitochondrial antigen identification
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In SLE, a homogeneous pattern is usually present, although SLE can also demonstrate
a speckled pattern. A nucleolar pattern is commonly seen in scleroderma. A
centromere pattern is common in CREST syndrome. Mixed connective tissue disease
(MCTD) is highly associated with a speckled ANA pattern.
IgM
IgA
IgG
IgE
Incomplete
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A decrease in which of the following in an AIDS patient are associated with increased
susceptibility to infection:
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CD4 positive lymphocytes, or T-helper cells, are infected and destroyed by the HIV
virus.
rheumatic fever
rheumatoid arthritis
lupus erythematosus
glomerulonephritis
lyme disease
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The ANA test, or anti-nuclear anitbody, is a screening test used to detect lupus
erythematosus. In this case the ANA test was positive. The RA test, or rheumatoid
factor, screens for the presence of an antibody linked with rheumatoid arthritis and
other conditions such as lupus erythematosus. Again, in this case, the RA test was
positive. Finally, a decrease in complement proteins indicates that the classic pathway
may have been activated resulting in immune complexes, a clinically significant
symptom of lupus erythematosus.
Question Difficulty: Level 6
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Multiplexing is the process of assaying several different genes within the same PCR
reaction.
True
False
A traveler is being evaluated at a local hospital for gastrointestinal pain, fever and
diarrhea. The patient has recently returned from a month-long visit to Southeast Asia.
The patient has no recent history of sexual activity or IV drug use but did describe
conditions of poor sanitation where they were visiting. If liver function test results
were elevated and bilirubinuria was noted, which of the following would be the next
logical test to order?
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The correct response is option D: ELISA testing for Hepatitis A antigens in the stool.
Hepatitis B, C and HIV are spread primarily through sexual contact or IV drug use
and needle sharing. While it is possible to contract these viruses through other means,
the patient history of visiting areas of low sanitation in Asia is more indicative of an
enterically (fecal-oral) transmitted organism like Hepatitis A. The earliest detectable
marker for Hepatitis A is the presence of viral antigens in the stool, which are most
often tested for using ELISA methodology.
What has happened in a titer, if tubes #5-7 show a stronger reaction than tubes #1-4?
poor technique
postzone reaction
prozone reaction
equivalence reaction
technical error
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If a cocktail that includes detergents, enzymes, and salts is used for DNA isolation,
which one of these ingredients will pull the DNA out from the sample?
Detergents
Salts
Enzymes
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If a cocktail that includes detergents, enzymes, and salts is used for DNA isolation, the
salts will pull the DNA out from the sample. The detergents break down the
membrane lipids and the enzymes remove the proteins.
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Cells do not only phagocytize bacteria, the can phagocytize other cells, cellular
remnants, and other waste products as well.
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A hapten is a low molecular weight molecule that can elicit an immune response only
when attached to a large carrier such as a protein.
A linear molecule
A lipid
Less than 5,000 molecular weight
Recognized as non-self
A haptene
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Only non-self antigens can be immunogenic. Self antigens are normally recognized by
the immune system as part of the host, so an immune response does not normally
occur. Non-self antigens are immunogenic since they have the potential to cause an
immune response.
Hay fever
Serum sickness
Arthus reaction
Anaphylactic shock
Tuberculin reaction
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The tuberculin reaction is a type of delayed hypersensitivity reaction that appears 48-
72 hours after antigen exposure, which is different from an immediate
hypersensitivity response, which generally appears within 12 minutes of an antigen
challenge. Arthus reactions and serum sickness are classified as a Type III
hypersensitivty reactions involving the formation of immune complexes. The other
choices above are all associated with immediate hypersensitivity reactions.
IgA
IgD
IgE
IgG
IgM
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Your answers are on the left. The correct answers are on the right and highlighted.
Nucleus A
Nucleolus B
Cytoplasm C
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Cultured T cells
Human plasma cells
Hybridomas
Cytotoxic T cells
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Monoclonal antibodies are monospecific antibodies that are the same because they are
made by one type of immune cell which are all clones of a unique parent cell, also
called a hybrid cell line, which usually arise from a hybridoma. The fusion of a
specific antibody-producing lymphocyte with a myeloma cell will multiply to become
a source of pure monoclonal antibody. This is often used in the manufacturing process
for monoclonal antibody reagents.
The DNA strand ending with a phosphate group is the 5' end.
True
False
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None
Testing for antibodies to the extractable nuclear antigens (ENA)
Testing for antibodies to dsDNA
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Speckled ANA patterns are usually followed-up with testing for antibodies to the
extractable nuclear antigens (ENA). Frequently this includes: Sm (Smith), U1-RNP,
SSA/Ro, SSB/La, and Scl-70.
Match terms with fragments resulting from papain digestion of an IgG molecule:
Your answers are on the left. The correct answers are on the right and highlighted.
Fc fragment B
Fab fragment A
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A = Fab fragment
B = Fc fragment
Which one of the following is not descriptive of the different branches of the immune
system?
Cell-Mediated
Extrinsic
Humoral
Innate
Non-specific
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Cell mediated, humoral, innate, and non-specific are all branches of the immune
system. Extrinsic is a branch of the coagulation cascade.
immunoglobulins
thymosin
serotonin
lymphokines
cytoplasmic granules
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T cells secrete lymphokines, which encourage cell growth, promote cell activation,
direct cellular traffic, destroy target cells, and initiate macrophages.
Which of the following antibody types is chiefly seen in the primary immune
response:
IgG
IgA
IgM
IgD
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IgM is the main immunoglobulin seen in the primary immune response. IgG is chiefly
seen in the secondary immune response. IgA is the predominant immunoglobulin in
body secretions. Finally, IgD is found in very small quantities and accounts for less
than 1% of the total immunoglobulin pool.
IgA
IgE
IgG
IgM
IgD
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Anti-human globulin (AHG) is created to bind with IgG antibodies at the Fc portion
of the antibodies. AHG will bind to both bound and unbound IgG antibodies. When
IgG is bound to RBCs, the AHG reagent will allow visible agglutination to occur.
Systemic sclerosis
Lupus erythematosus
Rheumatoid arthritis
Multiple sclerosis
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HBc antibody
HBc total
HBs antigen
Hbs antibody
Hbe antigen
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Hepatitis B surface antigen, or HBs, is a protein that is present on the surface of the
virus. The protein will be present via laboratory testing in the blood with acute and
chronic HBV infections
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Homogeneous and speckled is correct. In some labs this combination of patterns is the
most frequent of all positive ANAs.
Remember, however, any combination is possible and the other combinations listed
above are not uncommon.
2.3 days
2.8 days
5 - 6 days
18 - 23 days
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The half life of IgM and IgA are approximately 5-6 days. The half life of IgD and IgE
are 2.8 and 2.3 days, respectively.
The reaction that occurs when a soluble antigen is mixed with its specific antibody is
termed:
Precipitation
Agglutination
Sensitization
Fixation
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The reaction that occurs when a soluble antigen is mixed with its specific antibody is
termed precipitation. The visible precipitate is composed of large aggregates of
antigen crosslinked by antibodies.
Which T cell expresses the CD8 marker and acts specifically to kill tumor or virally
infected cells?
Helper T
T suppressor
T inducer/suppressor
T cytotoxic
Natural killer
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T-cells with the CD8 marker (also known as cytotoxic T cells) belong to a sub-group
of T lymphocytes that are capable of inducing the death of infected somatic or tumor
cells. Cytoxic T cells kill other cells that are infected with viruses or organisms as
well as other damaged or dysfunctional cells.
Myelin
Nuclear antigen
Surface antigens of bone marrow stem cells
Surface antigens of renal cells
Cytoplasmic processes
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The anti-nulear antigen test, or ANA test, is a test used to screen for autoimmune
diseases, especially systemic lupus erythematosus. It determines whether auto-
antibodies to cell nuclei are present. In 97% of cases diagnosed as SLE, patients have
a positive ANA test.
Cell Mediated
Specific
Humoral
Innate
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The innate immunity system is inherent and nonspecific; meaning that all pathogens
are attacked similarly. The skin, mucus in respiratory tracts, acid pH, and others are
all examples of the innate immunity system that the body has to prevent infection
upon first exposure.
The part of the immunoglobulin molecule that may be kappa or lambda chains is:
Fc
Fab
heavy chains
light chains
Hinge regions
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Light chains must always be either the kappa or lambda types in the formation of an
antibody.
You are performing a manual antibody titer in a clinical laboratory. You have made
serial dilutions of a patients plasma sample and you are looking for anti-streptolysin
O antibodies. The first tube contains pure patient plasma. The second tube contains a
1:2 dilution of the first tube. The third tube contains a 1:2 dilution of the second tube
and so on. The patients antibodies no longer give visible reaction in tube 5, indicating
their titer is tube 4 (or the last tube where there is a positive result). What is the
concentration of antibodies in tube 4 compared to the concentration in tube 1?
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Twelve weeks after onset of the disease, patients with uncomplicated acute hepatitis B
usually will demonstrate which of the following in their serum?
HBsAg
Anti-HTLV
Anti-HBe
Anti-HIV
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Jane Doe is a 39-year-old female who felt tired for several months, had pain in the
joints of her fingers, and recently developed a dermatitis following exposure to the
sun. The following test results were obtained on a blood sample drawn during the
initial evaluation:
Based on the clinical and laboratory findings, what disease should be suspected?
Rheumatoid arthritis
SLE
Sjogren's syndrome
Scleroderma
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The ANA titer is usually less than 1:160 in diseases other than SLE, so that the
possibility of SLE is increased in patients with higher titers. A speckled pattern is
commonly found in patients with SLE indicating binding to a variety of nonhistone,
small ribonucleoprotein (RNP) particles. A high crp level indicates that inflammation
is occuring, as in SLE for example, but does not indicate the location or cause of the
inflammation. Decreased complement levels also are associated with autoimmune
diseases. Both C3 and C4 levels are typically depressed in SLE
A 2 month old infant presented with history of chronic diarrhea and failure to thrive.
The child's mother also revealed that the baby has suffered from thrush (Candida
infections) since birth. Preliminary findings showed lymphopenia (<2,000),
hypogammaglobulinemia, and physical findings showed a small thymus, no tonsils or
lymph nodes. Which of the following immune deficiencies DOES the patient present?
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HDN is associated with alloantibodies from the mother's bloodstream directed against
the baby's red cell antigens.
Question Difficulty: Level 6
For the ANA test to be positive what must be present in the nucleus of the interphase
cell?
Strong staining
Centromeres
dsDNA
A clearly discernible pattern
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The strength of the reaction doesn't matter, only that it is clearly discernible.
Centromere and dsDNA are just some of the many different antigens, however,
without the corresponding (cognate) antibodies from the patient sample there is no
ANA pattern. Remember, to be ANA positive there must be a clearly discernible
pattern in the nucleus of the interphase cells.
Which of the following would you expect to find in the serum of a patient who has
recovered from Hepatitis B infection within 6 months after infection?
Anti-HBs
Anti-HBe
HBe
HBsAg
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The anti-Hbs antibody is produced in response to HBV surface antigen. The anti-Hbs
levels in the blood of patient who are infected with hepatitis B will rise during the
recovery phase. This test in the laboratory is used to detect previous exposure to HBV,
but can also be used to detect successful vaccination. This test is performed mainly to
determine whether or not vaccination is necessary or to determine if a person has
recovered from a hepatitis B infection and is immune.
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The correct response is option B: The Hepatitis B "e" Antigen (HBeAg). This antigen
indicates the virus is actively replicating and therefore the patient is very infectious.
The hepatitis B e antigen is present when the virus is actively replicating. In cases
of unintentional needlesticks, infectivity is of highest concern. The risk for infection is
greatest during phases of increased HBeAg serology. The Hepatitis B surface antigen
is the first detectable marker, but if the patient is known to have Hepatitis B already, it
would be relatively unhelpful to confirm the condition with another HBsAG test. The
core antigen is not detectable because it is covered by the nuclear envelope. Antibody
response patterns would not be very helpful either as the patient has already been
diagnosed with acute Hepatitis B. IgG antibodies would indicate recovery, which is
not the case for this patient and IgM antibodies indicating a recent or acute infection
would only confirm what is already known. Recall, in cases of unintentional
needlesticks, infectivity is of highest concern.
What is the MOST likely cause when clinical signs indicate HIV infection, but
antibody tests are negative?
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When clinical signs indicate HIV infection, but antibody tests are negative, a patient
can be in the "window phase" before antibody production. This means that the HIV
viral antigens have yet not produced an immune response great enough for laboratory
studies to detect anti-HIV antibodies. This window period can last up to 6 months
after infection.
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A specimen with a last reacting tube containing a 1:100 dilution, would be said to
have a titer of 100.
Which two stages of cell division are the most important for reading ANAs?
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While it is necessary to recognize all stages of cell division, the two most important
are interphase and metaphase. In order for the ANA to be positive there must be a
clearly discernible pattern in the nucleus of the interphase cells. Metaphase mitotic
cells are used to assist in identification of the ANA pattern.
The antinuclear antibody (ANA) test pattern that is seen in this image, viewed using
fluorescent microscopy, is characterized by smooth staining in the nuclei of the
interphase cells (a) and smooth staining in the chromosomal areas of the metaphase
mitotic cells (b). Which pattern is this?
Note: (a) points to the nuclei of two interphase cells. the nucleus of the interphase cell
is the primary consideration for discerning the ANA pattern. (b) indicates a metaphase
mitotic cell. Observing the chromosomal area and cytoplasm of the metaphase cell
may assist in identification of the ANA pattern.
Speckled
Centromere
SSA/Ro
Homogeneous
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Homogeneous is correct.
This pattern is characterized by smooth staining in the nuclei of the interphase cells
(a). The nucleoli may or may not stain. Notice the smooth staining in the
chromosomal areas of the metaphase mitotic cells (b).
In order for the ANA to be positive there must be a clearly discernible pattern in the
nuclei of the interphase cells. Metaphase mitotic cells are used to assist in
identification of the ANA pattern.
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A graft which is matched based on HLA and ABO typing is ideal. The mixed
lymphocyte culture, or MLC, should be negative also to prevent graft vs. host disease
in transplant procedures.
Which branch of the immune system has an immediate response on first exposure to a
foreign antigenic stimulus?
The correct answer is highlighted below
Cell Mediated
Specific
Humoral
Innate
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The innate immunity system is inherent and nonspecific; meaning that all pathogens
are attacked similarly. The skin, mucus in respiratory tracts, acid pH, and others are
all examples of the innate immunity system that the body has to prevent infection
upon first exposure.
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Though automated extraction machines have many benefits over manual methods the
costs are high and generally require a high throughput of samples in order to justify
the costs.
Automated extraction has many benefits over the traditional manual methods. The
most important benefit is that the nucleic acid isolated is constantly consistent. There
is a reduced amount of manipulation with dramatically decreases the chance of cross
contamination. Also, automated extraction machines are considered moderate
complexity and can be performed by a wider variety of laboratory professionals.
Question Difficulty: Level 6
Which of the following is the activation unit in the classical complement pathway?
C1
C2
C4, C2, C3
C5, C6, C7, C8, C9
C1, C2, C9
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Determining viral load is how one determines a positive diagnosis for HIV infection.
True
False
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Determining viral load is a way to monitor already infected individuals with HIV and
other viral diseases. The viral load can show how well treatments are working, show
potential medication resistance, and also indicate relapses.
B lymphocytes
Macrophages
Polymorphonuclear cells
T lymphocytes
Chemokines
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When an antigen comes in contact with the skin, the antigen is processed by cells in
the epidermis and come in contact with T lymphocytes. T lymphocytes recognize the
antigen as foreign and circulate through the bloodstream back to the epidermis and
produce an inflammatory response to eliminate the antigen, but this immune response
can produce a characteristic rash in the skin called contact dermititis.
Which one of the following would not be considered an anatomic barrier of the innate
immune system?
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The innate immunity system is inherent and nonspecific; meaning that all pathogens
are attacked similarly. The skin (dermis and epidermis), mucus membranes, acid (low)
pH, tears, saliva, cilia, mucous and others are all examples of the anatomic barriers of
the innate immune system which the body uses as its first line of defense. T
lymphocytes play a main role in the adaptive immune system and are not anatomical
barriers of the innate immune system.
The lymphokine most important for increased natural killer cell activity and decreased
viral replication in cells is:
Interferon gamma
Interleukin 3
Interleukin 4
Interleukin 5
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The greatest risk for the manifestation of anomalies in maternal rubella infection
during gestation is the:
first trimester
second trimester
third trimester
delivery
eighth month
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Infection early in the pregnancy is the most dangerous time period for the
manifestation of anomalies due to rubella. In fact, defects are rare when infection
occurs after 20 weeks gestation.
Your answers are on the left. The correct answers are on the right and highlighted.
Which immunoglobulin class is able to cross the placenta from the mother to the
fetus?
IgA
IgD
IgE
IgG
IgM
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Immunoglobulins of the IgG class are able to cross the placenta since they circulate as
monomers; thus they are small in size. The other types of immunoglobulins are too
large to cross the placental barrier, for example, IgM is a pentamer.
Basophils
Eosinophils
Polymorphonuclear neutrophils
Macrophages
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Your answers are on the left. The correct answers are on the right and highlighted.
Heavy chain A
Light chain B
Antigen binding occurs here C
Variable region D
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A = Heavy chain
B = Light chain
D = Variable region
Patient 1234 has a positive antinuclear antibody (ANA) test with a centromere pattern
at a 1:1280 titer.
Surprised by the result, the clinician calls and says the patient is only having mild
stiffness of her fingers and asks your opinion of the result. Which is your best reply?
The correct answer is highlighted below
You point out that centromere antibodies have been known to occur years prior
to the onset of the disease and the patient should be carefully examined and
monitored for the development of other symptoms.
You comment that it's common to over-read ANA results and since the ANA
testing is not diagnostic and prone to false results it's unlikely the results are of
any concern.
You state that the ANA results alone are diagnostic of systemic sclerosis and
since there is no cure for this disease the clinician is powerless to do anything.
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Centromere antibodies can precede the diagnosis by 10 to 15 years. While the ANA
alone is not diagnostic this patient is having clinical symptoms of stiff fingers that fits
nicely with the centromere ANA pattern. Early diagnosis and early treatment are
important to improve quality of life for the patient.
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Your answers are on the left. The correct answers are on the right and highlighted.
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Both IgG and IgM can activate complement. IgM is the largest immunoglobulin, as it
is a pentamer. IgG immunoglobulins have the ability to cross the placental barrier.
Finally, immediate hypersensitivity reactions are associated with IgE
immunoglobulins.
During which stage in real-time PCR can the fluorescent signal start to be recorded?
Plateau stage
Exponential stage
Leveling stage
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In real-time PCR the fluorescent signal is generally too low to detect until after the
first couple of cycles. However, detection usually occurs before the end of the
exponential stage.
Question Difficulty: Level 5
The immunoglobulin molecule is made up of both heavy and light chains - the light
chains can be comprised of which of the following:
Alpha or beta
Alpha or lambda
Kappa or beta
Kappa or lambda
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Only kappa or lambda chains can compose the light chains of an immunoglobulin
molecule.
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In real-time PCR, data are collected during nucleic acid amplification rather than at a
single endpoint.
Question Difficulty: Level 5
A cause of FALSE-POSITIVE results in the rapid plasma reagin (RPR) test for
syphilis is:
Gastroenteritis
Gonococcal urethritis
Infectious monocucleosis
Streptococcal pharyngitis
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The RPR test can show false positive results in the absence of syphilis if infectious
mononucleosis, SLE, antiphospholipid antibody syndrome, hepatitis A, leprosy, or
malaria are present.
Note: (a) points to the nucleus (nuclei) of the interphase cell(s), the primary
consideration for discerning the ANA pattern and (b) indicates the metaphase mitotic
cells. Observing the chromosomal area and cytoplasm of the metaphase cell may
assist in identification of the ANA pattern.
Pattern A
Pattern B
Pattern C
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In order for the ANA test to be positive there must be a clearly discernible pattern in
the nucleus of the interphase cells. Metaphase mitotic cells are used to assist in
identification of the ANA pattern.
Pattern C has no clearly discernable pattern in the nucleus of the interphase cells (a).
However, there is a discernable pattern staining in the cytoplasm (b). This pattern is
characteristic of anti-golgi antibodies.
ASO
anti-DNAse
M-antigen
Hemolysin
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hemolysins
opsonins
specific antitoxins
protective (neutralizing) antibodies
chemokines
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Radioimmunoassay (RIA)
Lateral Flow Immunoassay
Enzyme Immunoasay (ELISA)
Slide-based immunofluorescent assay (IFA) or Colorzyme
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Slide-based ANA tests using HEp-2 or HEp-2000 cells are the gold standard.
Dozens of different antibodies can be detected using slide-based assays. Solid phase
assays such as bead and ELISA assays lack sensitivity, resulting in an unacceptable
number of false negative results.
RIAand Lateral flow are not methods used for ANA testing.
How does the secondary immune response differ from the primary immune response?
All of the above choices are correct when differentiating the secondary immune
response differ from the primary immune response. In the secondary immune
response:
The antibody levels produced are higher in the secondary immune response
The lag phase (the time between exposure to antigen and production of
antibody) is shorter in the secondary
The primary immune response typically does not involve specific IgG
antibody
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The only statement that is not true of prozone is the last answer choice.
The remaining answers all describe the prozone effect. Prozone is the result of
antibody excess; it appears as a false negative, which becomes positive as the patient's
serum is diluted.
20oC
72oC
95oC
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Taq polymerase has optimal activity at 72C but can survive up to 95C.
IgE
T cells and macrophages
Nkcells and IgG antibodies
B cells and IgM antibodies
Epithelial skin cells
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When the tuberculosis (TB) skin test is performed, the T-lymphocyte cells with
receptor sites for tuberculosis antigens will produce a positive test result. The T-
lymphocytes secrete cytokines which attract macrophages to the site of the skin test.
Together these cells cause the swelling pattern to occur in the characteristic positive
skin test.
Question Difficulty: Level 7
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All lymphocytes originate, and are therefore derived from, hematopoietic stem cells
via hematopoiesis. The stem cells eventually mature into common lymphoid
progenitor cells. The progenitor cells then differentiate into their distinct lymphocyte
types. B cells mature into B lymphocytes in the bone marrow, while T cells migrate to
and mature in the thymus.
The prozone effect ( when performing a screening titer) is most likely to result in:
False positive
False negative
No reaction at all
Mixed field reaction
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Prozone effect (due to antibody excess) will result in an initial false negative in spite
of the large amount of antibody in the serum, followed by a positive result as the
specimen is diluted.
This antinuclear antibody (ANA) test is viewed using fluorescent microscopy. The
pattern is characterized by staining of the nucleoli in the nuclei of the interphase cells
(a). Which pattern is this?
Note: (a) points to the nuclei of the interphase cells, the primary consideration for
discerning the ANA pattern and (b) indicates a metaphase mitotic cell. Observing the
chromosomal area and cytoplasm of the metaphase cell may assist in identification of
the ANA pattern.
Nuclear lamins
SSA/Ro
Nucleolar
Large speckled
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Nucleolar is correct.
This pattern is characterized by staining in the nucleoli of the interphase cells (a). The
nucleolar staining can display subtle variations in staining inside the nucleoli
including smooth, speckled and clumpy. All are reported as ANA positive, Nucleolar.
In this sample staining is present in the chromosomal area of the metaphase mitotic
cells (b) along with some staining in the area outside of the chromosomal area. The
staining of the mitotics can be different with different anti-nucleolar antibodies.
Remember the ANA pattern is determined by staining in the interphase cells and the
mitotics are used to assist in interpretation.
The most common rapid slide test (MONOSPOT) for infectious mononucleosis
employs:
Horse erythrocytes
Sheep erythrocytes
Intact beef erythrocytes
None of the above
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A positive spot test shows agglutination of horse erythrocytes when added to patient
serum previously absorbed with guinea pig kidney, but not when added to patient
serum previously absorbed with beef erythrocyte stroma. The test is generally simple,
sensitive, and specific, but false negative are common in young children with
mononucleosis.
rheumatic fever
rheumatoid arthritis
lupus erythematosus
glomerulonephritis
lyme disease
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The ANA test, or anti-nuclear anitbody, is a screening test used to detect lupus
erythematosus. In this case the ANA test was positive. The RA test, or rheumatoid
factor, screens for the presence of an antibody linked with rheumatoid arthritis and
other conditions such as lupus erythematosus. Again, in this case, the RA test was
positive. Finally, a decrease in complement proteins indicates that the classic pathway
may have been activated resulting in immune complexes, a clinically significant
symptom of lupus erythematosus.
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In the humoral response, B-lymphocytes are stimulated to when an antigen binds to its
surface receptors. This sensitizes or primes the B cell and it undergoes clonal
selection, where it reproduces asexually by mitosis. Most of the family of clones
become plasma cells. These cells produce antibodies while other B cells become long-
lived memory cells.
The part of the molecule responsible for differences among immunoglobulin classes
is:
Fc
Fab
heavy chains
light chains
Hinge regions
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Antibodies are differentiated based on their heavy chains; different heavy chains
create different isotypes. Humans have five different isotypes of immunoglobulins.
C1
C4
factor D
C3
Factor B
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True
False
True
False
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The use of real-time PCR can detect differences as small as one base pair in a DNA
segment.
Corneal tissue may be transplanted successfully from one patient to another because:
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No HLA matching is performed for corneal tissue transplants as the cornea occupies a
privileged site not usually seen by the immune system. This term has been coined
"immune privileged".
The strength with which a multivalent antibody binds a multivalent antigen is termed
the:
Affinity
Avidity
Reactivity
Valence
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Fibronectin opsonizes bacteria and promotes their rapid phagocytosis. The remaining
choices are incorrect, as acute phase proteins NOT help to deactivate the complement
process, thiocyanate is NOT a secretory molecule found in skin secretions, and tumor
necrosis factor does NOT reduce phagocytosis at the site of an infection.
Question Difficulty: Level 6
IgG
IgA
IgM
IgE
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IgE levels are often increased in patients with allergic disease. IgE binds to the
membranes of mast cells and basophils, and if specific antigen is present to react with
the IgE molecule, degranulation of these cells occurs, releasing histamines, and other
substances into the blood or tissues.
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IgG
IgM
IgA
IgD
IgE
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active
passive
adoptive
innate
inactive
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Core antigen
Surface antigen
e antigen
Delta antigen
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Patients who have antibodies directed against the hepatitis B surface antigen, or anti-
HBs, have an immunity to hepatitis B. This is the principle behind hepatitis B
vaccinations as proteins associated with the hepatitis B surface antigen are
administered to an individual, who then produces an immune response against the
surface antigen. The antibodies produced through this process allows for the
immunization against the hepatitis B virus.
HbsAg - neg
HbeAg - neg
Anti-Hbc - neg
anti-HBs - pos
anti-HBe - neg
acute infection
chronic infection
immunization
susceptible
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Anti-HBs is expected to be positive post-vaccination. Since there are no other
antigens or corresponding antibodies present in the patient sample, it is likely that
patient has been immunized to hepatitis B and is no longer susceptible.
HbsAg - neg
HbeAg - neg
Anti-Hbc - neg
anti-HBs - pos
anti-HBe - neg
acute infection
chronic infection
immunization
susceptible
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Given the following results, what is the immune status of the patient?
HbsAg: positive
HbeAg: positive
Anti-HBc IgM: positive
Anti-HBs: negative
acute infection
chronic infection
immunization
susceptible
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HbsAg is positive in acute and chronic Hepatitis B infections, since the antigen is
found on the actual surface of the virus. HbeAg is present in the blood when the
hepatitis B viruses are replicating, indicating an active infection. Anti-Hbc IgM is
present due to the immune response to the presence of the hepatitis core antigen and
indicates an acute infection. Anti-HBs is generally interpreted as indicating recovery
and immunity from hepatitis B virus infection, according to the CDC.
The Western Blot Assay is used as a confirmatory test for which of the following:
HBsAg
CMV Antibody
Anti-HCV
Anti-HIV-1
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Genetic predisposition
Exposure to a triggering event
Low birth weight
Gender
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The amplification of DNA for northern blot is not an application of real-time PCR.
However, real-time PCR can be used to: diagnose infectious diseases, measure gene
transcription, measure drug therapy efficacy, and more. In contrast to regular reverse
transcriptase-PCR and analysis by agarose gels, real-time PCR gives quantitative
results. An additional advantage of real-time PCR is the relative ease and convenience
of use compared to some older methodologies.
There are two patterns present in this microscopic field from an antinuclear antibody
ANA test. The test is viewed using fluorescent microscopy. One pattern can be seen in
the interphase cells (a) and the area outside of the chromosomal area of the mitotics
(b). The other pattern is recognizable in the chromosomal area of the metaphase
mitotic cells (c). What are these two patterns?
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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.
Note: (a) points to the nuclei of several interphase cells, the primary consideration for
discerning the ANA pattern and (b) indicates a metaphase mitotic cell. Observing the
chromosomal area and cytoplasm of the metaphase cell may assist in identification of
the ANA pattern.
Homogeneous
Speckled
Nucleolar
Centromere
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