CRP and Hemolysis, Neutralization, and Complement Fixation Test

Originally thought to be an antibody to the C-polysaccharide of pneumococci

C-Reactive Protein

C reactive Protein is discovered by _________ and ______ in ________

Tillet and Francis in 1930

C reactive Protein is capable of the following:

O

A

P

CA

Opsonization

Agglutination

Precipitation

Complement Activation

CRP concentration:

Low:

Average:

High:

<1mg/dL

1-3 mg/dL

>3mg/dL

Determination of CRP is important in these process:

S

D

A

D

Screening

Diagnosis

Assessing

Differentiation

Procedure of CRP:

Bring ____________, ______________ control, ____________ control, and _________ to Room Temp

Prepare _________ of patient sample

Place _______ of Positive and Negative Control

On the third circle, put ___uL of _____________ sample

On the 4th circle put ___uL of _____________ sample

Positive, Negative, Serum sample

1:2 dilution of patient sampe

50uL of (+) and (-)

50uL of Undiluted serum

50uL of 1:2 diluted sample

Procedure of CRP (Part 2):

Add ____uL of ________ reagten to the sample

Mix with the stirrer, and place on the mechanical motor at ___-___ rpm for ____ mins

50uL of Latex Reagent

80-100 rpm for 2 mins

In CRP. examine macroscopically the presence or absence of visible _____________ immediately after removing the slide from the rotor

Agglutination

________________ - Distinct agglutination within 2 minutes indicates the presence of CRP in the sample

________________ - Smoot suspension without visible agglutination after 2 minutes

Positive Control

Negative Control

The approximate CRP concentration in the sample may be obtained by multiplying the CRP sensitivity ___mg/L with the last dilution factor with agglutination

6mg/L

It is the alteration, distribution of red blood cells in such a manner that hemoglobin is liberated

Hemolysis

Process of inactivation by the presence of antibodies that are known as neutralizing antibodies, antibodies that can inhibit the infectivity of a virus or the toxicity of a toxin molecule

Neutralization

The triggering of the classical complement pathway due to combination of antigen with specific antibody

Complement Fixation

Procedure, hemolysis Test:

Label tubes 1A to 6A and 1B to 6B

________ drops: 0, 1, 2, 3, 4, 5,

________ drops: 2, 2, 2, 2, 2,

Mix well, centrifuge for ____ seconds in the ________

Do not dispose the sediments, observe changes in the __________

Fresh Serum

RBC suspension

15 seconds in the serofuge

Supernatant

Hemolysis Test result
___________ - Pink to faint red supernatant

___________ - Clear, colorless supernatant

Positive

Negative

Complement Fixation Test Procedure:

Serum + Reagent 1:

Plus Reagent 2:

Plus Reagent 3

Bacterial Antigen

Complement Proteins

Indicator Red Cells (Sensitized RBCs)

Complement Fixation Test Result

_______________ - pink to faint red supernatant

_______________ - Clear, colorless supernatant

Negative

Positive