Mono & CMV

Rapid Agglutination Tests: Mono test kits examples

1) Monospot, 2) Mono slide. Both have an abosorption step with guinea pig kidney antigen and an agglutination step with Horse RBC which is more sensitive than sheep RBC

Paul Bunnell Screening Test: Serial Dilutions

1:7, 1:14, 1:28, 1:56, 1:112 dilutions made with Saline and sample. 0.1 mL of Sheep RBC suspension added to each dilution, mixed and observe for agglutination

Rapid Agglutination Tests: Procedure

2 reagents: Guinea pig antigen and Horse RBC. Add 1 drop of pt serum to 1 drop GP antigen, mix with paddle on card. This removes Foressman and Serum Sickness Ab. Then only non-Foressman remain. Add 1 drop of Horse RBC mix and rock & observe for agglutination (+ result)

Davidsohn Differential Tests (similar to Paul Bunnell Test)

2 titers: 1) Absorbed with guinea pig kidney antigens, 2) Absorbed with beef (ox) RBC.

Incubation of Mono lasts

4-7 weeks

CMV: Clinical Significance: Adults

40-90% adults carry CMV (asymptomatic

Heterophile tests: False negative

5-10% of adults and >50% of children less than 4 years old have NO heterophile antibodies

Mono: US Statistics

95% of those 35-40 yrs have been infected @ some point. 35-50% will have symptoms

Paul Bunnell Screening Test: Interpretation EBV

>28

Davidsohn Differential Tests: Guinea Pig Kidney Antigens

Absorb (remove) Ab from Foresman Group and Serum Sickness. Non - Forsmann remain in the supernatant after centrifugation. (discard RBC with other Ab at the the bottom). Serial Dilute and Add sheep RBC to both sets of titer.

Heterophile antigens

Ag that correlate with Heterophile Antibodies

Paul Bunnel Experimens: Horse and Sheep RBC react with which Heterophile Groups

All (Forssman Group, Non-Forssman Group, Serum Sickness)

Mono: Antibody that may or may not appear

Anti-EBNA (EBNA = Epstein Barr nuclear antigen). Show up in cells that have been infected when they die. Seen in RECURRENT infections.

EBV: Cancer

Burkitt Lymphoma and Nasopharyngeal carcinoma

CMV: Laboratory Testing: Tests for RAI

CMV coated well + pt serum w/CMV Ab--Incubated with Buffer with Urea and One w/o Urea. Wash both tubes. Add AHG labeled Enz, Add substrate, Colored product is directly proportional

CMV: Clinical Significance: Pregnant women (only if primary infection--contracted while pregnant)

Congenital CMV 2.5% of neonates that are affected by CMV. Disabilities may include hearing, vision, mental, liver, spleen, lungs, growth, bleeding (most will be blind and deaf)

Transmission of EBV

Contact with infected saliva (aka: Kissing Disease")

CMV: Laboratory Testing: Viral Culture

Done in research. After culture do a Direct Fluorescent Assay to detect Ag.

CMV: Latency (subclinical)

Dormant in myeloid cells. Hide in myeloid cells while still in the bone marrow.

Mono-Infectious Mononucleosis cause

EBV-Epstein Barr Virus

EBV specific serological tests are based on

ELISA (expensive). Usefulness is in a heterophile negative person or to differentiate between a primary and recurrent infection.

Mono Laboratory Result: Chem. -- Sever Liver infectoin

Increase in AST, ALT, LD. GGT and ALP and BIlirubin will also increase. Albumin and total protein will decrease.

Mono Laboratory Result: Hematology

Inverted ratio (lymphs higher than PMN). Lymphocytosis (high lymph). >/=15% Reactive lymph. Leukocytosis (increase WBC).

CMV: Laboratory Testing: RAI-Relative Avidity Index Interpretation

Low RAI - Primary Infection (result close to 0) High RAI - Recurrent Infection (result close to 1) Lab dependent cut off about interpretations.

How many reactive lymphs are seen in mono versus CMV

Mono: >/=15%, CMV <15% (maybe none in CMV)

CMV: Laboratory Testing: Indirect test: Latex Agglutination

Most used test to detect CMV

CMV: Laboratory Testing: Serological Markers

No name for antibody. IgM and then IgG created. IgG is low in primary infection and High in recurrent infection.IgM in created in primary infection.

Which Heterophile Antibody/Ag group is Mono part of

Non-Forssman Group

Paul Bunnel Experimens: Beef (Ox) RBC react with which Heterophile Groups

Non-Forssman Group and Serum Sickness (NOT Forssman Group)

Burkitt lymphoma

Non-Hodgkin Lymphoma that affects B-cell. Africa/Uganda have genetic mutation that make them prone to this cancer once infected by EBV. The EBV creates a cancerous mutation in the cell and then these cells replicate out of CTRL. Usually cancer goes to lymph node around throat.

Paul Bunnel Experimens: Human and Rabbit RBC react with which Heterophile Groups

None of the group

Paul Bunnell Screening Test: Interpretation Normal

Normal = 28 (we all have some heterophile antibodies).

CMV: Laboratory Testing: RAI calculation

RAI = Abs with Urea/ Abs w/o Urea (ex: 0.200/0.600 =1/3 =0.33 avitdity)

CMV: Laboratory Testing: Indirect hemagglutination

RBC covered in CMV Ag. Red Cross uses so that they can select seronegative donors for newborns, unborn babies (uterine transfusion), and immunocompromised.

CMV: Recurrent

Reactivated CMV infect lymph node and epithelial cells Then can go go Liver, Spleen, BM, Kidney, Lungs.

Mono: Convalescence

after acute disease person will feel better but not 100%. The virus goes dormanent. Virus goes dormant. Lasts 4-6 weeks.

CMV: Laboratory Testing: ELISA-VIDITEST

anti-CMV IgG and IgG avidity (used to calculated RAI)

Mono: Antibody that is second to appear

anti-EA (EA = early antigen). Starts in acute phase and decreases after convalesce period.

Mono: Antibody that is to 1st appear in Acute phase

anti-VCA (IgM that turns into IgG) can measure IgM or total. (VCA = Viral capsid Antigen). IgM is seen in Primary infections. IgG remain for life.

Must select CMV free blood products in blood banking for which populations

baby and immunocompromised.

ALP and GGT most affected when

biliary system is affected. These Enz stick out of the membrane in the biliary system so when bile comes through it will remove GGT and ALP and emulsify it so it will be absorbed in the circulatory system. Alcohol can also remove GGT

Mono: Lymphs can last a long time so may be infected in low numbers and evade the immune system

can become reinfected which will affect the liver only.

Mono: EBV goes into epithelial cells in the oropharynx through what receptors

cell receptor 2 = CD21 [Uses epithelial mechanism to replicate then burst cells resulting in a sore throat from a layer of destroyed epi in the oropharynx area]

Mono: Body activates Reactive lymphs which is what type of cell

cytotoxic T-cell to kill the virus in B-cell and throat. The Reactive lymph itself is NOT infected.

Nasopharyngyl cancer

has been connected to EBV in the USA

EBV Serological Tests: Done if we

have a False neg. (person doesn't produce heterophile antibodies) Then must use a serological tests specific for EBV. These are ELISA test that are very expensive.

Epstein Barr Virus is part of what family of viruses. What are other members of this family?

herpesviridae. Herpes Simplex I: cold sore and Herpes Simplex II: genital herpes are both part of this family in which the virus can become dormant and then will surface when immunity is decreased.

What is most sensitive to EBV antibodies (non-Forsemann)

horse RBC are capable of detecting agglutination at lower amt. of Ag and Ab

Mono Laboratory Result: Chem.--Liver is Infected

increase in AST and ALT and LD.

Heterophile Antibodies

most EBV tests are based on these type of Ab. These Abs are made against 1 species but have x-rxn with other species.

Davidsohn Differential Tests: Beef (ox) RBC

non-Foresman Group is absorbed and only Foresman and Serum sickness Ab remain. Serial Dilute and Add sheep RBC to both sets of titer.

EBV treatment

none--bed rest

CMV - Cytomegalovirus (may be written as HCMV) is transmitted

oral, sexual, organ transplant, blood transfusion, transplacental (main concern).

CMV: Clinical Significance: Immunocompromised

organ transplant or blood products with CMV

CD21

oropharynx epithelial marker and B cell marker. EBV may invade both of these cells through this recptor.

Davidsohn Differential Tests: is used

reference lab only. This is what Rapid mono tests are based on

Symptoms that CMV and Mono may have in common

sore throat, fatigue, fever, malaise

Reactive Lymph

stretched nucleus, cytoplams large, basophilia in peripherial or radial.

Mono: Acute Disease

symptoms include sore throat w/fever and malaise. Serology will be positive. Lasts about 2 weeks.

Age of ppl who get mono

teens and early adulthood is most common.

What is an important reason to diagnose EBV

to rule out other conditions

Paul Bunnel Experimens: Guinea Pig Kidney RBC react with which Heterophile Groups

Forssman Group and Serum Sickness (NOT to Non-Forssman Group)

Heterophile group

Forssman Group, Non-Forssman Group, Serum Sickness

Paul Bunnell Screening Test: False Positive

Forssman Group, Serum Sickness also react with Sheep RBC

Family of the CMV

Herpesviridae (just like mono)

Paul Bunnel Experimens: Which antigen react with Infectious mono (non-Forssman Group)

Horse, Sheep, Beef (Ox)

CMV: Laboratory Testing: ELISA

IgG and IgM

Paul Bunnell Screening Test: Reporting

Reported as the inverse of the highest dilution to show agglutination.

CMV: Laboratory Testing: Molecular Dianositics

Research only for Direct test of Ag

Heterophile tests: False positive

Rubella, CMV, viral heptitis, malaria, autoimmune. Pancreatic cancer.

CMV: Primary Infection

Sore throat, fever, swollen glands (esp. neck), fatigue. Mononucleosis features. Elevated liver enzymes may or may not be seen. Reactive lymph but not as high as mono.