Wednesday, October 24, 2007

Reticulocyte Count

Hi all.
I am kinda lost with the shedule on whoose gona blog this week or what, so i'll just post something okay (: sorry to the one who ought to blog this week! HA.

Okay, i shall introduce Recticulocyte Count Test that is done in the Haematology Section;

1) Recticulocyte Count (RC) (in another words, rectic count)

Introduction: The recticulocyte count is used in the evaluation of anemia as it accurately reflects the amount of erythrocytes production taking place in the bone marrow.
Relationship btwn anaemic condition and erythrocyte production in bone marrow (in normal cases):
anaemic condition= increase in RBC production therefore increase of RC in blood
However, if the RC is not raised, it shows an indictation of impaired bone marrow function or lack of eythrocytes stimulus.

Principle: Recticulocytes are junvenile red cells. Thus they contain remnants of the ribosome and the ribonucleic acid, which are present in larger amt in the cytoplasm of the nucleated precursors from they are derived.
New Methylene Blue or Brilliant Cresyl Blue are supravital dyes that are used to measure reticulocytes.
Currently, there are 2 methods being employed:
a)Manual Method: The use of Brilliant Cresyl Blue
b) Automated method: Reticulocytes Package by Cell Dyn Ruby analyzer
We used the automated method cause its more efficient and contributes to a faster Turn Around Time (TAT).

Procedure of Automated Method

1) When using the retoculocyte reagent, verify the expiration date and store the stock reagent in the dark at room temperature
2) Label patient accession No. on to the tube of reticulocyte reagent
3) Verify that the whole blood specimen is warmed to room temperature and well mixed pior to
sampling
4) Pipette 20 uL of the blood sample into each labelled tube of reticulocyte reagent
5) Incubate the stained Reticulocyte specimens on a rotator or in a rack, after fully inverting the stained specimens 4-6 times. Inbubation must be performed according to the reagent package insert.

Note: The stained Reticulocyte specimens must incubate for at least 15mins but no more than 2 hours pior to processing on the Cell Ruby machine.

Although the process is time-efficient, there are some limitations of the procedure.

Okay im done with elaborating. Hm i shall list some definations of defined abnormalities (that we 'kinda' need to know and understand in the haema section)


Leukocytosis is an elevation of the white blood cell count (the leukocyte count) above the normal range. The normal adult human leukocyte count in peripheral blood is 4.4-10.8 x 109/L. A white blood count of 11.0 x 109/L or more suggests leukocytosis.
Leukocytosis is very common in acutely ill patients. It occurs in response to a wide variety of conditions, including viral, bacterial, fungal, or parasitic infection, cancer, hemorrhage, and exposure to certain medications or chemicals including steroids. Leukocytosis can also be the first indication of
neoplastic growth of leukocytes.
For lung diseases like pneumonia,tuberculosis etc. WBC count are very inportant for the diagnosis of the disease that means leucocytosis can be seen in above mentioned diseases


Neutrophilia is a condition where a person has a high number of neutrophil granulocytes in their blood.
Neutrophils are the primary
white blood cells that respond to a bacterial infection, so the most common cause of marked neutrophilia is a bacterial infection.
Neutrophils are also increased in any
acute inflammation, so will be raised after a heart attack or other infarct.
As well as increasing in number, neutrophils show other changes in infection and inflammation.
A neutrophilia might also be the result of a
malignancy. Chronic myelogenous leukemia(CML or chronic myeloid leukaemia) is a disease where the blood cells proliferate out of control. These cells may be neutrophils. Neutrophilia can also be caused by appendicitis.

Lymphopenia is the condition in which there exists an abnormally low number of
lymphocytes in the blood.
Lymphopenia can be caused by various types of
chemotherapy, such as with cytotoxic agents or immunosuppresive drugs. Some malignancies in the bone marrow will also cause lymphopenia.
A decreased number of lymphocytes (notably
T cells) is present in those with AIDS. People exposed to large doses of radiation, such as those involved with Chernobyl can also exhibit a lymphopenia.
Lymphopenia may be present as part of a
pancytopenia, when the total number of blood cells are reduced. This can occur in marrow failure.

Monocytosis is an increase in the number of circulating
monocytes. In humans, 950/μL is regarded as at the upper limit of normal; monocyte counts above this level are regarded as monocytosis.
Monocytosis often occurs during
chronic inflammation. Diseases that produce this state:
Infections:
tuberculosis, brucellosis, listeriosis, subacute bacterial endocarditis, syphilis, infectious mononucleosis and other viral infections and many protozoal and rickettsial infections (e.g. kala azar, malaria, Rocky Mountain spotted fever).

Eosinophilia is the state of having a high concentration of eosinophils (
eosinophil granulocytes) in the blood. The normal concentration is between 0 and 0.5 x 109 eosinophils per litre of blood. Eosinophilia can be reactive (roughly, allergic) or non reactive.
Diseases that feature eosinophilia:
Hypereosinophilic syndrome, Parasitic infections, allergic disorders.
The release of interleukin 5 by T cells, mast cells and macrophages stimulates the production of eosinophils
Anemia is a deficiency of
red blood cells (RBCs) and/or hemoglobin. This results in a reduced ability of blood to transfer oxygen to the tissues, causing tissue hypoxia.
The three main classes of anemia include excessive blood loss (acutely such as a hemorrhage or chronically through low-volume loss), excessive blood cell destruction (hemolysis) or deficient red blood cell production (ineffective hematopoiesis).

Basophilia is an uncommon cause of leukocytosis. Basophils are inflammatory mediators of substances such as histamine. These cells, along with similar tissue-based cells (mast cells), have receptors for IgE and participate in the degranulation of white blood cells that occurs during allergic reactions, including anaphylaxis
Causes; Infections: viral infections (varicella), chronic sinusitis Inflammatory conditions: inflammatory bowel disease, chronic airway inflammation, chronic dermatitis
Thrombocytosis is the presence of high platelet
counts in the blood, and can be either reactive or primary (also termed essential and caused by a myeloproliferative disease).
High platelet counts can occur in patients with
polycythemia vera (high red blood cell counts), and is an additional risk factor for complications.

p.s. the definations above are just F.Y.I okaaay.

See yall soooon! (:

Natalie
TG01

13 comments:

The Lab Freaks said...

Hello Nat,

Erm, ask u ah.. What u mean by supravital dyes? Hopefully how does the New Methylene Blue or Brilliant Cresyl Blue stain the recticulocytes?

Thanks anyway!

Royston
TG01

VASTYJ said...

Hi Nat,

Are the reagents pre-packaged into individual tubes? What will happen if the sample is incubated for more than 2 hours?

Ying Ying
TG01

The Lab Freaks said...

Nat!

What are the limitations of this test?

see ya soon!

charmaine
tg01

MedBankers said...

hey there!

regarding RC, do you do periperal blood film after adding the brilliant cresyl blue?

cheers!
elaine

The Lab Freaks said...

Nat!!
Eh, Im also curious to know what supravital dye means. Another thing, I find it strange that you do Retic count in this manner at your lab. At my lab, we do a PBF and then stain it. I guess thats what you meant by manual method right? Cool.

Sharifah
TG01

ALsubs said...

Hello Natalie,
why do you have to incubate the blood sample? Thank you.

Shu Hui
TG02

first6weeks said...

Hi Nat,

Whats the advantages/disadvantages of using the automated method as opposed to using the manual method?


Desmond Heng
0503179D
TG02

we are the XiaoBianTai-7! said...

hi natalie,

from your blog i understand that your lab do automatic count for retic but i would like to know does your lab do manual count for retic? and under what kind of situation.

Lizzie TG01 0503194C

royal physicians said...

Hello!

I will just like to know does the lab still use the reagents that just expired? Or they are thrown away straightaway? Thanks!!

kangting
0503331A

The Lab Freaks said...

hello nat!

how much time does this automated method save compare to the manual one? and is this method cost effective?

~Jeremy~
TG01

The Lab Freaks said...

Hello to all. sorry for the late delay of answering yall qns.

Hi Royston,
Supravital dyes are of, relating to, or capable of staining living cells after their removal from a living or recently dead organism.
The Brilliant Cresyl Blue/New Methylene Blue will stain the reticulocytes and cause it to appear greenish blue with deep intracellular precipitate. The dye exerts a stablizing effect on the RBCs.

Hi Ying Ying,
Yes the reagents are all pre-packed into individual tubes in a box.
If the sample is incubated for <2hours, the results will be invalid as the RBCs might not be stablized for accuracy of results.


Hi Charmaine,
The limitations are:
1) Difficulty in differeniating reticulocytes from cells containing "Pappenheimer bodies"
2)Cell Dyn Ruby TM method is a nucleic acid staining method. Therefore otehr substances that contain nucleic acid cld potentially be enumerated as rectics.
3) RC can be increased with pregnancy.
4)If tourniquet is applied on the arm too long, (i.e. >1min) may cause inaccurate results.

Hi elaine:
erm yes we do, but that is mainly for the manual method which we seldom use here!


Hi sharifah!
erm you can look at my ans for roy's for your ans! and yes, its the manual method.

Hi shu hui,
The incubation time is for the reaction to take place; staining of the immature red cells with the brilliant cresyl dye.


Hi Desmond,
Advantages are it is cost-effective(large production), high accuracy of results and most of the procedure are automated thus lowers the rate of errors(human).
As for disadvantages, you could probably look at my ans for charmaine? its rather the same as limitations! ha. ouh. erm one major disadvantage is that there are alot of interfernces that is able to affect the result e.g. nucleated RBCs, Hemolysis, Parasites.

Hi Lizzie,
Yes we do do manually. We do it only when the machine has a flag on a certain specific sample, or if the result of the RC of a sample is too high.


Hello Kangting,
Urm, as far as i know of, we do not use any reagents are past the expiry date for, it may cause inaccurate test results.


Hi jeremy!
urmmmm i cant really determine how much time saved compared the manual and automated cause frankly speaking, the incubation time of both methods already takes up 15mins. hmm perhaps the automated is faster by a few mins but <5mins? lol but the automated has a higher accuracy of results thou.
And yes, it is cost-effective when there is a large amt of RC tests requested.


Finally i've completed all the qns! (its really alot! HA)
Thanks for asking, hope yall will be able to understand the brief answers! (:

cheers,
natalie.

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