ID- Lab Testing and Diagnostics

Question 1

Signal amplification has reduced false-positive results from cross contamination because the number of target molecules is not altered.

Answer 1

Consequently, the signal is directly proportional to the amount of target sequence present in the original specimen.

Question 2

Lab procedures can be useful tools for three things

Answer 2

1. Diagnosis of infection
2. Assess the stage of infection or infectious process
3. Evaluate HAI

Question 3

What are the two key elements in selection and implementation of a test method?

Answer 3

1. Accuracy
2. Precision

Question 4

What is accuracy and what two terms can be used to describe it?

Answer 4

Accuracy is the closeness of the result obtained to the true value.
1. Sensitivity
2. Specificity

Question 5

What is sensitivity?

Answer 5

1. The ability to detect all true cases of the disease
2. The absence of of false-negatives.
   #true positives/ (#true positives + # false negatives)

Question 6

What is specificity?

Answer 6

The ability to correctly identify a negative result. The absence of false positive results.
# true negatives/ (True negatives + False positive results)

Question 7

What is precision?

Answer 7

Precision= repeatability. When repeat testing of same sample produces same or similar results.

Question 8

What is a proficiency testing program?

Answer 8

An external evaluation of the quality of a laboratory’s performance.

Question 9

3 Common purposes for tests include:

Answer 9

1. Screening
2. Confirmation
3. Diagnosis

Question 10

What is screening used for?

Answer 10

Used to identify a disease process in individuals without signs or symptoms of the disease. In general they have high sensitivity, but require confirmation due to low specificity.

Question 11

What is confirmation used for?

Answer 11

Used after a positive screen to ensure accuracy of the result and confirm presence of the condition.

Question 12

Diagnosis

Answer 12

used for the evaluation of persons suspected of having a given disease state or characteristic.

Question 13

How can definitive diagnosis be made (4)

Answer 13

1. visualization of the agent in a specimen.
2. Detecting specific products from an infectious agent (CDI Toxin)
3. Detecting an immunological response (Ag-Ab) (Hep B surface antibody)
4. Detecting presence of agent through nucleic acid hybridization and amplification techniques.

Question 14

Direct Examination Methodologies include (3)

Answer 14

1. Gram Stain
2. Histology/Cytology
3. Wet Mount

Question 15

Antigen Detection

Answer 15

1. Helpful for early diagnosis where cultures not yet positive, possible or practical.
2. Direct method to test for infectious agents
3. They differ from direct visualization in that they Include immunologic or serologic procedures
4. Methods designed to detect entire agent (virus) or part of the agent (bacterial cell wall). (ELISA)

Question 16

Antibody

Answer 16

Antibody immunoglobulin Produced by B-lymphocytes.
Facilitate the removal of the agent. Humeral.
5 Classes of antibodies- each with different properties and functions

Question 17

Antibody Detection

Answer 17

Indirect Method by assessment of host response
Results may be qualitative (+/-) or quantitatively (titres)- should be interpreted in consideration of the predictive value of the specific test.
Consider historical infection.

Question 18

Molecular Diagnostic Testing

Answer 18

subject of false +, sensitive to contamination
(PCR)
1. Target Amplification
2. Probe Amplification
3. Signal Amplification- signal directly proportional to the amount of target sequence present in the original specimen (reduced false +)

Question 19

Tests for infectious process include:

Answer 19

1. Body Fluid Analysis: total protein, specific gravity, cell count, boy fluid glucose, gram stain, culture.
2. Cerebrospinal Fluid Analysis: color and clarity, protein, glucose, and WBCs.

Question 20

What is the preferred method of differential cell count?

Answer 20

Centrifugation-> allows Wright’s staining of air dried cytospins and has good cell yield and preservation of cells.

Question 21

C-reactive Protein (CRP)

Answer 21

Abnormal serum glycoprotein produced by liver during acute inflammation. Usually disappears quickly when inflammation subsides.
Sometimes used in diagnosis of meningitis, pneumonia, sepsis, TB, UTI.

Question 22

Cold Agglutinins

Answer 22

Antibodies that cause clumping of type O red blood cells at cold temperatures. M. pneumonia, viral pneumonia, primary atypical pneumonia.

Question 23

Liver Function Tests

Answer 23

Help aid in diagnosis of infectious processes such as chronic syphilis, subacute bacterial endocarditis, TB, inflammation due to certain viral infections (Hep b, C).

Question 24

Arterial Blood Gas

Answer 24

Determines gas exchange across alveolar-capillary membrane in the lungs.
Worsening gas exchange may indicate inflammation in lung from bacterial or viral infection.
ABGs critical components in ID of HA pneumonias and ventilator associated pneumonias.

Question 25

The WBC count is the total number of WBCs (leukocytes) in 1mm3 of peripheral blood

Answer 25

WBC count > 10 000 usually indicates infection, inflammation, or leukemic neoplashia.

Question 26

Neutraphil

Answer 26

Primary function= phagocytosis. Acute bacterial infections stimulate immature neturophilis (shift to left). Appendicitis.

Question 27

Lymphocytes

Answer 27

increases associated with pertussis, syphilis, toxoplasmosis. Lymphocytes with frothy cytoplasm (atypical lymphs) associated with viral infections: cytomegalovirus, mono).

Question 28

Monocytes

Answer 28

Increased monocytes commonly associated with with EBV, TB, subacute bacterial endocarditis, syphilis, protzoan.

Question 29

Eosinophils

Answer 29

typically associated with allergic reactions. May be increased in parasitic infections, leprosy, tb, systemic fungal infections, scarlet fever, chlamydial pneumonia in infancy.

Question 30

Basophilia

Answer 30

Uncommon finding can be found in food/drug allergies, variola/varicella infections, ulcerative colitis.

Question 31

Complete Blood Count: Absolute Neutrophil Count

Answer 31

Neutropenia is reduction of ANC to 1.5-2 x109/L. ANC= Total WBCx ((Segs +Bands)/100) Severe Neutropenia (agranulocytosis)= <0.5. Determine whether an immunocompromised patient should be placed in protective isolation

Question 32

Lymphocyte Subset : 2 types

Answer 32

T cells (mature in thymus)- involved in cellular-type immune reactions. B cells (bone marrow) humoral activity- antibodies T cells= killer cells CD8= suppressor cells CD4 Helper cells

Question 33

Sedimentation Rate

Answer 33

Rate at which RBCs or erythrocytes fall. Lack sensitivity and specificity for disease processes; however, increased ESR rate associated with acute infection and inflammation.

Question 34

Fecal Leukocytes

Answer 34

Determine whether diarrhea is invasive or non-invasive to the mucosa of the colon. Presence of leukocytes indicates the cause of diarrhea is an organism or process, such as Salmonella, Shigella, Amoeba, Campylobacter, helicobacter, yersinia is breaking mucosal barrier of the colon. Not common in viral enteritis or toxin-mediated diarrhea. CDI may or may not be positive.

Question 35

Exotoxin

Answer 35

Proteins secreted by gram (-/+) (CDI, Clostridium tetani, Corynebacterium diphtheriae, staph aureus). Many have A/B motif. A active portion, B binding portion. Ability to lyse host cells, alter function of host cells, resulting in death of cell. Stimulates T-Cell responses in a cytokine cascade. EIA and HPLC to detect.

Question 36

Endotoxins

Answer 36

lipid A section of lipoolysaccharides that are only found on the outer membrane of some gram negative organisms (Ecoli, salmonella, shigella, pseudomonas, Neisseria, H Influenza, Bordetella pertussis, vibrio cholerae). Cause damage to host through cytokine production by macrophages. Limulus amebocyte lysate LAL test-> used to test water quality for hemodialysis.

Question 37

Weil-Felix Agglutinin

Answer 37

To detect and differentiate rickettsial antibodies in serum. Used to diagnose: Rocky Mountain spotted fever, Q fever, epidemic typhus, murine typhus, scrub typhus, rickettsialpox.

Question 38

Urinalysis

Answer 38

Color, Clarity, presence of proteins, glucose, ketones, blood, nitrate, leukocyte esterase. Dipstick: presence of leukocyte esterase and nitrite- screening tool for infection. Nitrite- bacteria produce an enzyme that reduces urinary nitrates to nitrites. leukocytes >10WBC/mm3 unspun or urine, or >5 WEBC high power field of spun urine. pyuria can be helpful when differentiating between colonization and true infection in the presence of bacteriuria.