Trinity University of Asia College of Medical Technology

SIM100 Lecture Handouts (Prof. Jude Anthony Trinidad, RMT, MSMT, ASCPi CM)
de Guzman, Angelo Christian O. | 3MT01

BACTERIAL SEROLOGIC TESTS  Detection of Group A Streptococcal Antibodies

BACTERIA AND IMMUNE DEFENSES o The most diagnostically important Antibodies:
 Immune defense against bacteria  Anti-Streptolysin O (ASO)
o Lysozyme  Anti-DNAse B – Most common
o Humoral Factors  Anti-NADase
o Phagocytosis  Anti-Hyaluronidase
o Humoral Immunity o Importance: Diagnose the Acute Rheumatic Fever and
o Cell-mediated Immunity Post Streptococcal Glomerulonephritis
 Bacterial ways to inhibit the immune system  ASO Testing
o Avoiding Antibody o Neutralization reaction
o Blocking Phagocytosis o Dilution of patient serum, to which a measured amount
 Inhibition of chemotaxin of Streptolysin O reagent is added
 Inhibition of opsonin o Titer – Reciprocal of the highest dilution demonstrating
 Capsule no hemolysis
o Inactivating complement cascade o Units: Todd Units/IU
o Performed by Nephelometric Methods
GROUP A STREPTOCOCCI  Anti-DNAse B Testing
o Highly specific for Group A streptococcal sequelae
o Neutralization reaction
 (+) Without clearing
o Detect patients with Acute Rheumatic Fever
 Streptozyme testing
o Excellent screening test
o Slide Agglutination test

Helicobacter pylori
 Agent of Gastric and Duodenal Ulcer
 Based on the Lancefield classification (which is based on  Strong Urease Producer
Antigen class) o Produces urease to counteract the acidity of the stomach
 Streptococcus pyogenes  Spiral bacterium
o β-Hemolytic  Virulence Factor:
o M and T Protein o CagA
 M Protein – inhibits C3b (inhibits the o Vacuolating Toxin (VacA)
complement pathway)  Antigen detection procedures
o Streptolysin o Endoscopy
 Streptolysin O – Heat-labile; Antigenic o Culture: Most specific
 Streptolysin S – Heat-stable; non-antigenic o Gastric Biopsy
 Clinical Manifestation o Urease Biopsy test
o Streptococcal Toxic Shock Syndrome o PCR
o Upper Respiratory Tract Infection – Pharyngitis  Detection of Helicobacter pylori Antibodies
o Skin Infection – Impetigo o IgG, IgM, IgA
o Scarlet Fever – Erythrogenic Toxins  IgG – Primary Antibody detected
 Exotoxin A, B, C o Primary screening detection
 Damaging Sequelae
o Damage done after infection with Streptococcus pyogenes Mycoplasma pneumoniae
 Acute Rheumatic Fever – complication after  Class Mollicutes
sore throat/Pharyngitis  Eatons Agent
 Post Streptococcal Glomerulonephritis – follow  Leading cause of upper respiratory infection worldwide
after infection of either skin or pharynx  Pleomorphic – no cell wall but with Sterol (Required for growth)
o The Anti-M Protein can attack the heart and kidneys  Culture: Fried Egg Appearance
because the Antigen of the Streptococcus and of the  Causes Walking Pneumonia
heart and the kidney shares the same epitope – cross-  Detection of Mycoplasma pneumoniae by culture
reactivity o Transport media
 Detection of Group A Streptococcal Antigen  Trypticase soy broth with 0.5% Albumin
o Culture (Bacitracin)  SP4 Medium
o Enzyme Immunoassay  Viral Transport Medium
o Latex Agglutination o Delay in plating – frozen at -70°C
o PCR  Detection of Antibody to Mycoplasma pneumoniae
o Cold Agglutinin that reacts with the I/i Antigen

BS Medical Technology | Third Year, Second Semester | March, 2015 | Clinical Immunology and Serology Semi-Final Period |Page 1 of 4
 Trinity University of Asia College of Medical Technology
SIM100 Lecture Handouts (Prof. Jude Anthony Trinidad, RMT, MSMT, ASCPi CM)
de Guzman, Angelo Christian O. | 3MT01

 Ii Blood Group – Blood group where Humans – Accidental Hosts except Rickettsia prowazekii
Mycoplasma pneumoniae reacts with  Serological Diagnosis
o Enzyme Immunoassay, Immunofluorescence Assay o Weil-Felix (Agglutination)
o PCR, Nucleic Acid Amplification o IFA, IBA (Immunoblot Assay), Micro-IF
o *Need tissue culture
FEBRILE AGGLUTININ o Gold Standard – IFA, Micro-IF
 Antibody demonstrated in Microbial diseases that are  Weil-Felix Interpretation
manifested by high fever Rickettsial disease OX-19 OX-2 OX-K
o Typhoid – Salmonella typhi Scrub Typhus - - ++++
o Typhus – Rickettsia RMSF
o Brucellosis – Brucella Epidemic Typhus ++++ + -
Marine Typhus
o Tularemia – Francisella tularensis
Q Fever - - -
 Harder to culture – so use Brucella Ag to detect
Rickettsial pox - - -
Tularemia – cross reactivity
 Use Proteus Antigen – non specific  Cross reactivity
 Detects Antibodies
o OX-19 (Proteus vulgaris)
o OX-2 (Proteus vulgaris)
TYPHOID FEVER S
 Salmonella typhi o OX-K (Proteus mirabilis)
 Acquired by ingestion
LYME DISEASE
 Typhoid Mary
 Causative Agent: Borrelia burgdorferi
 Salmonella Antigen
 Transmitted via Ixodes spp.
o H Antigen and K Antigen
 Laboratory diagnosis
 Thermolabile
o Immunofluorescence
 H Antigen (Houch) – Flagellar
o EIA
 K Antigen (Kapsel) – Capsular
 Vi – Virulence Antigen o Western Blot
o O Antigen o PCR
 Thermostable  Bull’s Eye Rash
 (One) Somatic/Body Antigen o Erythema cronicum migrans – Rash in Lyme disease
 Laboratory Diagnosis of Typhoid Fever
o Culture Method – Gold Standard LEPTOSPIROSIS
 Causative Agent: Leptospira interrogans
o Widal Test – Detect Agglutination
 Infection stages
 (+) Agglutination
o Septicemia stage
o Typhidot
o Immunological stage
 Detect IgG (Past infection) or IgM (Present
 Laboratory Diagnosis
infection)
o Culture Method
 Samples
o 1st and 2nd weeks – Blood o Microscopy
o 3rd week – Urine o Serological Test
o 4th week - Stool
SYPHILIS/GREAT POX/EVIL POX/SPANIS DISEASE
 Caused by Treponema pallidum subsp. pallidum
RICKETTSIAL INFECTION
 Corkscrew motility
 Treponema pallidum subsp. pallidum
o Treponema pallidum subsp. pallidum – Syphilis
o Treponema pallidum subsp. pertenuae - Yaws
o Treponema pallidum subsp. endemicum – Nonvenereal
(endemic) syphilis/Bejel
o Treponema carateumi – Pinta
o Species are indistinguishable from each other
 General characteristics of Treponema pallidum
o Too thin to be seen with light microscopy in specimens
 2 Distinct groups: stained with Gram stain or Giemsa stain – Dark field
o Spotted Fever Group (SFG) microscopy or Fluorescence Microscope
o Typhus Group (TG) o Intracellular pathogen
 Intracellular, Obligate (Need host to live) o Has Treponemal rare outer membrane proteins
 Gram negative, short rods, coccobacillus (TROMPS) – delays immune system of host
 Associated to the Arthropod Vector  Mode of transmission
o Biological Vector o Sexual intercourse

BS Medical Technology | Third Year, Second Semester | March, 2015 | Clinical Immunology and Serology Semi-Final Period |Page 2 of 4
 Trinity University of Asia College of Medical Technology
SIM100 Lecture Handouts (Prof. Jude Anthony Trinidad, RMT, MSMT, ASCPi CM)
de Guzman, Angelo Christian O. | 3MT01

o Parental exposure  Principle: Flocculation

o Congenital infections during pregnancy  Reagent: Cardiolipin, Lecithin, Cholesterol
 Clinical Manifestations  Cardiolipin – Main reagent to detect
o Primary regain
 Appearance of Hard Chancre  Lecithin – Neutralizes the anti-
 Laboratory Tests complementary properties of
 Dark Field Microscopy (Presence of Cardiolipin
coiled organism with Corkscrew  Cholesterol – Provides Adsorption
motility)  Rotator: 180 rpm for 4 minutes (Serum VDRL),
 Early infection – No Antibody production 180 rpm for 8 minutes (CSF VDRL)
 Sample: Lesions  Ring Diameter: 14mm (Serum VDRL), 16mm
o Secondary (CSF VDRL)
 Characterized by generalized rash and lesions  Depth – 1.75 mm
 Appearance of Condylomata lata  Qualitative VDRL – Detect presence or absence
 Laboratory Tests of Reagin (Use Gauge 18)
 Dark Field Microscopy  Quantitative VDRL – Measures the
 Serologic Test concentration of the Reagin Antibodies
 Sample: Fluid from lesions  Gauge 19
o Latent  Syringe used: Humilton’s Syringe
 Generally after 2nd year of infection  CSF VDRL – Gauge 21 or 22
 Patient is Asymptomatic – not infectious  Reporting
 The only indicator of infection is a positive  Non-reactive – no clumps
serologic test  Weakly reactive – Small clumps
o Tertiary  Reactive – medium to large clumps
 Characterized by destructive lesions known as  False (+) VDRL Results – SLE, RF, IM, Malaria,
Gummas Pregnancy
 3 Manifestations o Rapid Plasma Reagin (RPR)
 Gomatosyphilis  Uses unheated serum, results are read
 Cardiovascular macroscopically
 Neurosyphilis  Principle: Flocculation
 Congenital Syphilis  Reagent: colorless alcoholic solution containing
o Hutchinsonian triad Cardiolipin, Lecithin, Charcoal (for easier
 Hutchinson’s teeth visualization), Choline Chloride, and Thimerosal
 Interstitial keratitis (Preservative)
 Nerve deafness  Rotator: 100 RPM for 8 minutes
o Other characteristics  Ring Diameter: 18 mm
 Fissuring around the mouth and anus  Antigen delivery needle: Gauge 20 needle, 60
 Skeletal lesions drops/mL
 Perforation of the Palate and Collapse of nasal  Use Humilton’s syringe
bones  Saddle-nose deformity  Serologic Test for Syphilis
 Laboratory diagnosis o Treponemal Serologic Test/Specific Methods
o Direct detection of Spirochetes  Uses true Treponemal Antigen – detect
o Non-treponemal serologic test Treponemal Antibodies
 Screening confirmed by treponemal serological o 2 sources:
test  Non-pathogenic Reiter strain
o Treponemal serological test  Pathogenic Nichol’s strain
 Specific method o FTA-ABS
 Diagnostic Markers  Principle: Indirect Fluorescent Immunoassay
o Reagin Antibodies  Direct Relationship
 Aka Wasserman reagent/Phosphatidyl Glycerol  Used with Dark Microscopy
 Antigen used: Cardiolipin (From the heart of a  Reagent Antigen: Nichol’s strain dried and fixed
cow) on slide
 Non-treponemal marker  Absorbent: Reiter treponemes
o Anti-treponemal Antibodies  Removes cross-reactivity with other
 Treponemal marker treponemes
 Antigen used is Treponemal Antigen o Treponema pallidum Immobilization test (TPI)
o VDRL (Venereal disease research laboratory)  Principle: The Antibody produced against
 Uses heated serum/CSF (Inactivated serum) Treponema pallidum plus complement can
 Result is read microscopically immobilize the 5 treponemes

BS Medical Technology | Third Year, Second Semester | March, 2015 | Clinical Immunology and Serology Semi-Final Period |Page 3 of 4
 Trinity University of Asia College of Medical Technology
SIM100 Lecture Handouts (Prof. Jude Anthony Trinidad, RMT, MSMT, ASCPi CM)
de Guzman, Angelo Christian O. | 3MT01

 Reagent Antigen: Live actively motile

Treponema pallidum organisms (extracted from
the lesions of infected Rabbits)
 (+): >50% Immobilizaed Treponemes

BS Medical Technology | Third Year, Second Semester | March, 2015 | Clinical Immunology and Serology Semi-Final Period |Page 4 of 4