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short case study I need help to answer short questionssee the attachment
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You are selected to be a key member of an assay development team at your local hospital.
There has been an uptick of infections with a newly discovered spirochete. Physicians have
requested that an in-house serology assay to detect if someone has been previously infected
with this microbe. Researchers have discovered that the Z protein/antigen is very unique and
specific to this pathogen. You have been assigned to develop assays that can detect IgG
antibodies to this Z antigen.
1. Fortunately, researchers have already purified Z protein/antigen and have successfully
artificially adsorbed the antigen on a carrier latex bead. Which technique would be the
most useful: Direct Agglutination Assay, Passive Agglutination Assay, or Reverse
Passive Agglutination Assay? (1 pt)
2. Describe the principle of the assay you indicated in question 1. Indicate the steps
involved, the analyte, how the assay detects the analyte (e.g. purchased/kit antibodies or
antigens), and how to interpret the results? (3 pts)
3. One of your known undiluted “positive” samples does not cause agglutination for the
assay you have chosen. However, once you dilute the sample, agglutination is
observed. Explain the phenomenon that is occurring in this example (hint: think zone of
equivalence). (2 pts)
4. Your team is asked to develop a more sensitive ELISA assay. Z antigens/protein are
successfully adsorbed onto a microtiter plate. Knowing this, which technique would be
most useful for the detection of IgG antibodies against Z protein in a patient sample:
Sandwich ELISA (capture), Competitive EIA, or classical non-competitive EIA? (1 pt)
5. Describe the principle of the assay you indicated in question 4. Indicate the steps
involved, the analyte, how the assay detects the analyte (e.g. purchased/kit antibodies or
antigens, enzyme labelled antigens or antibodies, etc), and how to interpret the results?
(3 pts)
A 26 year-old male presented to urgent care complaining of open sores on his mouth
and penile area. He indicated having multiple sexual encounters with different
individuals over the past year. The physician ordered several STD tests. The rapid
plasma regain (RPR) test was reactive/positive. The HIV rapid was negative, and PCR
Panel for gonorrhea, chlamydia and trichomonas was also negative.
1. Based on patient’s history and symptoms, what disease do you suspect the
patient most likely has? (1 pt)
2. What is the causative microbe of this disease (genus and species)? (1 pt)
3. In addition to the RPR, what is another screening test that can be utilized to
detect this disease? (1 pt)
4. Because the RPR was positive, does that confirm the patient had the disease
indicated in question 1? If not, indicate a confirmatory test that could be ordered.
(2 pts)
5. If the RPR was negative, should the physician order a confirmatory test? (1 pt)
6. What stage of this disease do you suspect the patient to be in? (1 pt)
7. What would be the best treatment option for the patient? (1 pt)
A 46-year old female complained at a recent appointment of stiffness and pain in her
wrists, hands and fingers that had worsened over the past year. Serum was collected,
and a series of autoimmune tests were ordered. The fluorescent antinuclear antibody
results were positive (low titer) with a speckled pattern. Anti-SM and Anti-dsDNA ELISA
tests were negative. The rheumatoid factor (RF) test was positive, and her c-reactive
protein (CRP) levels were above normal.
1. Based on the results and history, what autoimmune disease does the patient
most likely have? (1 pt)
2. What is RF, and does the presence confirm the autoimmune condition you
indicated in question 1? (2 pts)
3. As an MLS, what ELISA test would you suggest the physician ordered to confirm
the disease indicated in question 1 (ie what antibody would be more specific for
this condition). (1 pt)
4. What type of hypersensitivity does this condition induce? Briefly explain your
answer. (2 pt)
5. What does high levels of CRP indicate in patients? (1 pt)
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