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Immunology and Serology [LAB] > Laboratory Activities 8, 9 and 10 – Non-specific tests for Syphilis: VDRL and RPR > Flashcards

Laboratory Activities 8, 9 and 10 – Non-specific tests for Syphilis: VDRL and RPR Flashcards

1
Q

is a contagious sexually transmitted disease caused by the spirochete Treponema pallidum subspecies pallidum.

A

Syphilis

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2
Q

This spirochete causes an immune response in the body by producing antibody-like substance in plasma or serum known as reagin.

A

Syphilis

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3
Q

[?], a phospholipid derived from beef heart, reacts with reagin, and this antigenic property is used in non-Treponemal tests.

A

Cardiolipin

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4
Q

NON-TREPONEMAL TESTS AND TRUE-TREPONEMAL TESTS

A
  1. Wasserman test
  2. Direct Detection of Spirochetes
  3. Serologic Tests
  4. DNA probe
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5
Q

Direct Detection of Spirochetes

A

• Darkfield microscopy
• Direct immunefluorescence

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6
Q

Non-Treponemal Serologic Tests

A

1) Venereal Disease Research Laboratory (VDRL)
2) Rapid Plasma Reagin (RPR)

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7
Q

Treponemal Serologic Tests or True Treponemal Tests

A

1) Treponema pallidum Immobilization (TPI) test
2) Fluorescent Treponemal Antibody Absorption Test (FTA-ABS)
3) Enzyme immunoassay
4) Hemagglutination Tests

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8
Q

DNA probe

A

a. Polymerase chain reaction (PCR)
b. Southern blot

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9
Q

SYPHILIS
Caused by:

A

Treponema pallidum subspecies pallidum (originally called Spirochaeta pallida)

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10
Q

SYPHILIS
Transmission:

A

Sexual contact, direct blood transfusion or transplacental route

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11
Q

SYPHILIS STAGES

A
  1. Primary syphilis
  2. Secondary syphilis
  3. Latent syphilis
  4. Tertiary syphilis
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12
Q

• Associated lesion lasts from 1-6 weeks, during which time the lesion heals spontaneously

A
  1. Primary syphilis
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13
Q

• Generalized lymphadenopathy, malaise, fever, pharyngitis, and a rash often involving the mucous membranes and the skin

A
  1. Secondary syphilis
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14
Q

• Lack of clinical signs and symptoms

A
  1. Latent syphilis
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15
Q

• Manifested by gummatous syphilis, cardiovascular disease, and neurosyphilis

A
  1. Tertiary syphilis
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16
Q

o The spirochete is transmitted from mother-to-fetus

A

Congenital syphilis

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17
Q

Congenital syphilis
o Manifested by the Hutchinsonian triad:

A

notched incisor teeth, keratitis, and deafness

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18
Q

Congenital syphilis
o Best to use a test that detects IgM antibodies

A
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19
Q

o Detection of IgM antibodies that are produced by the infant’s immune system

A

Congenital syphilis

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20
Q

Congenital syphilis
o Treatment:

A

Arsphenamine or Salvarsan or compound 606 Penicillin

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21
Q

Wasserman test

First diagnostic blood test for syphilis developed in 1906 based on the serologic principle of

A
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22
Q

o Also known as cardiolipin; identified as a phospholipid (diphosphatidyl glycerol)

A

Wasserman antigen

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23
Q

o Derived from the host or extract from beef heart

A

Wasserman antigen

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24
Q

derived from extracts of liver from newborns that had died of congenital syphilis

A

Wasserman antigen

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25
Q

demonstrated that tissues from animals without syphilis, mainly beef heart extracted in alcohol, could be used equally well as antigens

A

Landsteiner

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26
Q

• Darkfield microscopy o Positive result:

A

Corkscrew motility

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27
Q

o Uses anti-treponemal antibody with fluorescent tag against Treponema pallidum antigen in a patient

A

• Direct immunefluorescence

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28
Q

Non-Treponemal Serologic Tests
o Determine the presence of ____________________

A
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29
Q

Non-Treponemal Serologic Tests
o Based on the principle of ____________________

A
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30
Q

Non-Treponemal Serologic Tests
Examples:

A

• Venereal Disease Research Laboratory (VDRL)
• Rapid Plasma Reagin (RPR)
• Toluidine Red Unheated Serum Test (TRUST)
• Unheated Serum Reagin (USR)
• Reagin Screen Test (RST)

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31
Q

Non-Treponemal Serologic Tests
False-positive: 10-30% may be biologic false positives

A

infectious mononucleosis, infectious hepatitis, malaria, leprosy, lupus erythematosus, rheumatoid arthritis, advance age, pregnancy

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32
Q

Non-Treponemal Serologic Tests
Positive:

A

Treponemal infections

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33
Q

Causes yaws: chronic non venereal disease of skin and bones

A

Treponema pallidum subspecies pertenue

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34
Q

Causes bejel: lesions in the oral cavity, oral mucosa, skin, bones, and nasopharynx

A

Treponema pallidum subspecies endemicum

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35
Q

Causes pinta: ulcerative skin disease

A

Treponema carateum

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36
Q

Transmission: traumatized skin comes in contact with an infected lesion

A

Treponema pallidum subspecies pertenue

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37
Q

Transmission: mouth to mouth by utensils

A

Treponema pallidum subspecies endemicum

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38
Q

Transmission: traumatized skin comes in contact with an infected lesion

A

Treponema carateum

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39
Q

Both a qualitative and quantitative slide flocculation test for serum and cerebrospinal fluid (CSF)

A

Venereal Disease Research Laboratory (VDRL)

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40
Q

Venereal Disease Research Laboratory (VDRL) Antigen consists of:

A

• 0.03% cardiolipin: main reacting group
• 0.9% cholesterol: enhance the reacting surface of cardiolipin
• 0.21% lecithin: removes anti-complement activity of cardiolipin

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41
Q

main reacting group

A

• 0.03% cardiolipin:

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42
Q

enhance the reacting surface of cardiolipin

A

• 0.9% cholesterol:

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43
Q

removes anti-complement activity of cardiolipin

A

• 0.21% lecithin:

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44
Q

Aside from the VDRL antigen, (?) is also employed

A

buffered saline containing sodium chloride, formaldehyde, secondary sodium phosphate, and primary potassium phosphate

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45
Q

o Rotation:
• VDRL serum _____ rpm for _____ minutes
• VDRL CSF _____ rpm for _____ minutes

A
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46
Q

Venereal Disease Research Laboratory (VDRL)
Examination for flocculation:

A

Microscopic

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47
Q

o 18 gauge needle without bevel that will deliver 60 drops of antigen suspension per ml

A

Qualitative Serum VDRL

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48
Q

o 19 gauge needle without bevel that will deliver 75 drops of antigen suspension per ml

A

Quantitative Serum VDRL

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49
Q

o 23 gauge needle that with or without bevel that will deliver 100 drops of saline per mL

A

Quantitative Serum VDRL

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50
Q

o 21 or 22 gauge needle that will deliver 100 drops per ml

A

• CSF VDRL

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51
Q

Rapid Plasma Reagin (RPR)
o Reagent: Antigen is similar to the VDRL antigen with the addition of the following:

A

• Charcoal: allows macroscopic visualization
• EDTA: prevents oxidation
• Thimerosal: preservative
• Choline chloride: inactivates the complement

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52
Q

allows macroscopic visualization

A

• Charcoal:

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53
Q

prevents oxidation

A

• EDTA:

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54
Q

preservative

A

• Thimerosal:

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55
Q

inactivates the complement

A

• Choline chloride:

56
Q

RPR
o Specimen

A

serum

57
Q

RPR
o Rotation: _____ rpm for _____ minutes

A
58
Q

RPR
o Examination for flocculation:

A

Macroscopic

59
Q

RPR
o Antigen delivery needle:

A

• 20 gauge disposable needle without bevel, 60 drops are obtained in 1 ml

60
Q

Detect Treponemal antibodies

A

Treponemal Serologic Tests or True Treponemal Tests

61
Q

Involves mixing of patient serum with live, actively motile Treponema pallidum extracted from testicular chancre of a rabbit

A

Treponema pallidum Immobilization (TPI) test

62
Q

o Patient serum is heat-inactivated and made with a sorbent consisting of nonpathogenic treponemes (Reiter strain) which removes the cross-reactivity with treponemes other than Treponema pallidum

A

Fluorescent Treponemal Antibody Absorption Test (FTA-ABS)

63
Q

o Nichols strain of Treponema pallidum has been fixed to slides used for the test

A

Fluorescent Treponemal Antibody Absorption Test (FTA-ABS)

64
Q

For detection of anti-treponemal antibody from a patient

A

Enzyme immunoassay

65
Q

Antigen: RBCs sensitized with Nichols strains

A

Hemagglutination Tests

66
Q

Hemagglutination Tests

A

• Hemagglutination Treponemal Test for Syphilis (HATTS)
• Treponema pallidum Hemagglutination Assay (TPHA)
• Microhemagglutination Assay for Antibodies to Treponema pallidum (MHA-TP)

67
Q

o Reagent: Sheep RBCs coated with antigens from Nichols strain of Treponema pallidum; Sorbent

A

Hemagglutination Tests

68
Q

Hemagglutination Tests
o Positive reaction:

A

Rough or jagged pattern of cells in microtiter wells

69
Q

• Patient DNA matched with treponemal DNA

A

DNA probe

70
Q

• A means of amplifying tiny quantities of nucleic acid using a heat-stable polymerase enzyme and a primer that is specific for the DNA sequence desired

A

Polymerase chain reaction (PCR)

71
Q

Technique for the identification of specific DNA sequences in which DNA is cleaved into fragments by enzymes, separated electrophoretically, denatured, transferred to a nitrocellulose membrane, and incubated with a labeled probe that is specific for the sequence of interest

A

Southern blot

72
Q

 is a protein found in blood plasma that shows increased levels in response to inflammation.

A

C-reactive protein (CRP)

73
Q

 It is synthesized in the liver and has a half-life of 20 to 30 hours.

A

CRP

74
Q

 It belongs to a group of proteins known as acute phase reactants whose concentrations increase during a general, nonspecific response to infectious and inflammatory processes, cellular necrosis, and malignant neoplasia.

A

CRP

75
Q

 contributes to non-specific immunity by activating complement and stimulating phagocytosis.

A

CRP

76
Q

A nonspecific, acute-phase, reactant glycoprotein that increases very significantly with the onset of a substantial inflammatory stimulus, such as infection, myocardial infarction, or surgical procedures

A

C-reactive protein (CRP)

77
Q

CRP Common serologic test:

A

Latex agglutination

78
Q

(?) are reacted with patient serum. In this case, the CRP is acting as the antigen. If CRP is present above normal threshold levels, the antigen antibody combination will result in a (?)

A

Latex particles coated with antibody to CRP
visible agglutination reaction

79
Q

sensitive, although nonspecific, indicator of inflammation

A

elevated CRP level

80
Q

Normal levels range from __________ in newborns to __________ in adults

A
81
Q

CRP

gives a positive result with the undiluted specimen

A

0.6 mg/dL or higher

82
Q

CRP levels can also be used to monitor the outcome of

A

surgery, graft rejection, drug therapy with anti-inflammatory agents, and recurrence of malignancies

83
Q

 is a chronic disease that affects the joints characterized by inflammation, stiffness, and soreness.

A

Rheumatoid arthritis (RA)

84
Q

 Most individuals affected by (?) experience a gradual onset of the disease, but usually, all joints in the body will be affected.

A

rheumatoid arthritis

85
Q

 As Rheumatoid arthritis progresses, other organs of the body can develop an

A

inflammatory condition.

86
Q

 The determination of the rheumatoid factor is essential in the (?) of rheumatoid arthritis

A

prognosis and therapeutic management

87
Q

• An anti-antibody that binds to the Fc portion of abnormal IgG
• Associated to rheumatoid arthritis

A

Rheumatoid factor (RF)

88
Q

o Chronic systemic inflammatory disorder in which joint cartilage, ligaments, and tendons are destroyed

A

rheumatoid arthritis

89
Q

o An autoimmune disease that affects the synovial membrane of multiple joints

A

rheumatoid arthritis

90
Q

Also noted in chronic hepatitis, syphilis, systemic lupus erythematosus (SLE), scleroderma, Sjogren’s syndrome, and Bcell lymphoproliferative disorders

A

Rheumatoid factor (RF)

91
Q

• Principle:
• Passive agglutination can be employed to test for the presence of the antibody, using a carrier particle such as (?) with IgG. When (?) is mixed with the sensitized reagent cells, visible agglutination occurs.

A

sheep erythrocytes sensitized
patient serum containing RF

92
Q

Latex fixation test by

A

Singer and Plotz

93
Q

Latex agglutination titer lower limit based on an undiluted sample and diluted sample at

A

1:10

94
Q

• Very low titer (present also in other diseases such as SLE, endocarditis, tuberculosis, syphilis, viral infection, and diseases of the liver, lung, or kidney)

A

o If positive only in undiluted

95
Q

• Low titer can be found in approximately 1 percent of healthy individuals

A

o If positive only in undiluted

96
Q

• RF is present in the specimen at a level generally associated with RA

A

o If positive at 1:10 dilution

97
Q

• Interpretation of results based on titer:
o Positive: ____________________
o Weakly positive: ____________________
o Negative: ____________________

A
98
Q

Rheumatoid factor (RF) Methods

A
  1. Latex agglutination
  2. Sheep cell agglutination test: Rose-Waaler Test
  3. Sensitized alligator erythrocyte test: Cohen et.al
  4. Bentonite flocculation test: Bloch and Bunim
99
Q

 is useful in the diagnosis of Streptococcal infection and its relative effects on organs, especially the heart and kidneys.

A

Anti-Streptolysin O (ASO) test

100
Q

 Though the causative agent of Group A Streptococcal infection can be cultured, it can also be detected through serologic techniques.

A

Anti-Streptolysin O (ASO) test

101
Q

is a gram-positive coccus responsible for several human infections, some of which can have serious sequelae

A

Streptococcus pyogenes

102
Q

: Major virulence factor for Streptococcus pyogenes; more than 60 M serotypes have been identified

A

o M protein

103
Q

• Virtually all strains of Streptococcus pyogenes produce two hemolysins

A

o Bacterial toxins

104
Q

• Enzyme that causes hemolysis by binding to cholesterol in the RBC membrane

A
  1. Streptolysin O (SLO)
105
Q

• Enzyme that causes hemolysis by disrupting the selective permeability of the RBC membrane

A
  1. Streptolysin S
106
Q

caused by Streptococcus pyogenes include cellulitis, impetigo, and erysipelas

A

Skin infections

107
Q

caused by Streptococcus pyogenes are characterized by fever, sore throat, and pharyngeal edema

A

Upper respiratory tract infections (URTI)

108
Q

caused by a strain of Streptococcus pyogenes that produces an erythrogenic toxin, which results in a characteristic rash; fever and sore throat are also present

A

Scarlet fever

109
Q

a complication seen following upper respiratory tract infections

A

Rheumatic fever (RF)

110
Q

o All M serotypes that cause pharyngitis have been implicated in

A

RF

111
Q

o results in damage to heart valves, and patients with rheumatic heart disease have an increased risk of developing endocarditis and other cardiac problems in later years

A

RF

112
Q

may occur after pharyngitis or skin infections

A

Poststreptococcal glomerulonephritis

113
Q

o Only a few M serotypes cause this type of glomerulonephritis; patients have an increased risk of developing renal failure later

A

Poststreptococcal glomerulonephritis

114
Q

• The mechanisms by which these (?) occur are not fully understood

A

sequelae

115
Q

• Antibodies to streptococcal cell membranes cross-react with (?) in cardiac muscle cells, which results in cell damage

A

myosin

116
Q

• Antigen-antibody complexes form at the glomerular basement membrane and attract inflammatory cells that cause

A

renal tissue damage

117
Q

During infection with Streptococcus pyogenes, SLO produced by Streptococcus pyogenes elicits an immune response, and specific antibodies are formed→ these antibodies neutralize the

A

hemolytic activity of the SLO

118
Q

The ASO titer begins to increase approximately (?) and peaks (?)

A

7 days after infection
after 4 to 6 weeks

119
Q

SLO is added to serial dilutions of patient serum, along with (?) as indicator cells → if the patient serum contains antibodies against SLO, the antibodies will complex with the corresponding antigens → these complexes block the hemolytic activity of the antigen, and no hemolysis occurs

A

group O RBCs

120
Q

• The ASO titer is reported as the reciprocal of the highest dilution that shows no hemolysis and is expressed in

A

Todd units

121
Q

o Normal: titer of __________ Todd units

A
122
Q

o Moderately elevated: titer at least (?) in an adult and (?) in a child

A

240 Todd units
320 Todd units

123
Q

• High titers with Streptococcal infections, rheumatic fever, glomerulonephritis

A

Anti-streptolysin O (ASO) titer

124
Q

Based on the neutralization of the hemolytic activity of streptolysin O

A

Conventional method:

125
Q

Conventional method: Positive Resul
Serum with Ab (ASO) → + streptolysin O reagent → neutralization of streptolysin O by patient Ab (ASO) → + RBC (indicator) → _________________________ (presence or absence of hemolysis?

A
126
Q

Conventional method:

Negative Result

Serum without Ab (no ASO) → + streptolysin O reagent ® neutralization of streptolysin O by patient Ab (ASO) → + RBC (indicator) → _________________________ (presence or absence of hemolysis?)

A
127
Q

Conventional method Controls:

A

 RBC control (RBCs + buffer) should show NO hemolysis
 Streptolysin control (streptolysin O + buffer + RBCs) should show hemolysis

128
Q

Rheumatoid factor (RF)

Common methods:

A

 Nephelometry
 Latex agglutination

129
Q

sometimes appear earlier than Anti-streptolysin O in streptococcal pharyngitis

A

Anti-DNase B

130
Q

 are increased in the 15% to 20% of RF patients who do not have elevated ASO titers

A

Anti-DNase B levels

131
Q

 Sensitivity is increased for the detection of glomerulonephritis preceded by streptococcal skin infections as this type of disease does not stimulate ASO antibodies

A

Anti-DNase B testing

132
Q

Measurement is based on neutralization methodology

A

Anti-DNase B testing

133
Q

If anti-DNase B antibodies are present, they will neutralize reagent DNase B, preventing it from depolymerizing DNA → presence of DNase is measured by its effect on

A

DNA-methyl green conjugate

134
Q

Slide agglutination screening test for detection of antibodies to several streptococcal antigens

A

Streptozyme testing

135
Q

Principle: Sheep RBCs are coated with antigens so that antibodies to any of the streptococcal antigens can be detected

A

Streptozyme testing

136
Q

 Several rapid tests are now available to detect streptococcal antigens
 The advantage is their (?) over other test methods; however, a significant number of false-negative results occur

A

speed

Immunology and Serology [LAB] (16 decks)

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