Slides on "compatibility test and how you give blood," revised and corrected in April 2010 available at:
http://www.box.net/shared/1pkr80l22t
Wednesday, April 29, 2009
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compatibility tests and how to allocate
Slides on "compatibility test and how you give blood," revised and corrected in April 2010 available at:
http://www.box.net/shared/1pkr80l22t
Slides on "compatibility test and how you give blood," revised and corrected in April 2010 available at:
http://www.box.net/shared/1pkr80l22t
Tuesday, April 21, 2009
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Immediate Spin Cross-proof "fast" at room temperature: Determination
Try crusade "fast" at room temperature: Immediate Spin
essentially serves to confirm the AB0 compatibility between recipient and donor. Should always be done, especially if you do not routinely double check .(....)
Technique:
> mix 3 drops of serum / plasma of the recipient + 1 drop of red blood cells donor in suspension at 3-4% in saline.
> read for agglutination.
Even this test may be performed in test tubes (centrifugation and read out as above) or slide / plate, slide, however, this survey is less reliable and requires more time and a closer reading eventually with the aid of a microscope at low magnification.
Interpretation:
Absence of agglutination: the recipient and the donor are confirmed compatible with respect to the AB0 group.
· Presence of agglutination: mean incompatibility (ABO or other antibodies that may be cold or not clinically significant) NOT transfuse, gruppaggio check a possible error in the direct evidence and assess the possible presence of cold auto-antibodies (see Indirect determination ABO). 
Try crusade "fast" at room temperature: Immediate Spin
essentially serves to confirm the AB0 compatibility between recipient and donor. Should always be done, especially if you do not routinely double check .(....)
Technique:
> mix 3 drops of serum / plasma of the recipient + 1 drop of red blood cells donor in suspension at 3-4% in saline.
> read for agglutination.
Even this test may be performed in test tubes (centrifugation and read out as above) or slide / plate, slide, however, this survey is less reliable and requires more time and a closer reading eventually with the aid of a microscope at low magnification.
Interpretation:
Absence of agglutination: the recipient and the donor are confirmed compatible with respect to the AB0 group.
· Presence of agglutination: mean incompatibility (ABO or other antibodies that may be cold or not clinically significant) NOT transfuse, gruppaggio check a possible error in the direct evidence and assess the possible presence of cold auto-antibodies (see Indirect determination ABO).
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AB0 indirect or "rebuttal"
Determination AB0 indirect or "rebuttal"
It 's a test complementary and not alternative to direct determination. Its usefulness is mainly limited to the confirmation of the AB0 determined using commercial antisera allowing to interpret the possible, though infrequent, questionable results and to highlight the very rare false negatives and false positives of direct evidence .(....)
The counter is usually performed at the first determination group, while for the future control us is limited to the direct determination.
The indirect test for the presence of A and Anti B antibodies that are naturally present if the fees are absent antigens. To test should have known A and B red cells suspended in fresh saline to 3-4%. The red cell suspension may be stored in the refrigerator for several days. In practice this means having in the area known donors of group A and B available to frequent small blood samples.
Technique:
> mix 2 drops of serum or plasma from blood typing + 1 drop of red blood cells suspended in A or B notes to 3-4% in saline.
> read for agglutination
Even this test can be performed in test tubes (centrifugation and read out as above) or slide, slide, however, this survey is less reliable and requires more time and a closer reading eventually with the aid of a microscope at low magnification.
Expected Results:
° samples / plasma of group A and B red cells agglutinate 0 ° samples
/ plasma of group A red cells agglutinate B
° samples / plasma of group B red cells agglutinate A
° samples / group AB plasma did not agglutinate A or B red blood cells or red blood cells
If you are faced with a discrepancy between direct evidence and rebuttal, and that we are faced with the unexpected positive or negative, will need to make a series of additional tests to explain the reason for the discrepancy. In addition to a simple error in identification / execution of the test are different, but rare, situations that may explain unexpected results, by sub-groups AB0 rare (see the A2 subgroup [1] ) in the presence of autoantibodies cold [ 2] . In these cases to clarify the presence of indirect agglutination test is useful to the unexpected execution of a self [3] .
In general in the case of a recipient whose group AB0 found a discrepancy between direct and indirect evidence should be explained not only transfuse blood donor certainly 0.
Babies and up to 4 months of age do not yet have natural anti-A antibodies or anti-B, in these cases the proof is so often negative and its execution is merely confusing. Unexpected indirect negative test can also be observed in elderly or immunocompromised, in these cases, the absence or weakness of the reactivity is due to the low titer of regular.
A test of indirect confirmation of ABO compatibility of donor and recipient groups is represented by the cross-test at room temperature.
[1] Occasionally the plasma / serum A subgroup of A2 agglutinate red blood cells called type A1 is the most common. These clumps have no clinical significance, therefore the cells A1 can be safely transfused to patients with group A2 in spite of the apparent "incompatibility" [2] Presence of autoantibodies cold: in the case of agglutination of both cells ( A and B) which does not correspond to the direct determination, as expected, a group can be 0 in the presence of autoantibodies active at room temperature (antibodies or Fedde crioagglutinine). In these cases it is useful to add also a test of self-control in such cases will be positive, contrary to what is normally expected. If self-control is positive it is highly likely the presence of cold autoantibodies (that agglutinate at room temperature almost all the cells). These are generally not significant, that is not cause transfusion reactions. In case of positive self-repeating all the tests at 37 ° C by incubating cells and serum / plasma in a water bath. If the agglutination of self disappears (that is no longer active cold autoantibodies) can interpret the residual agglutination with red cells A or B or with those of the donor (during the test cross) as significant. If that persists in the agglutination test cross will be prudent not to transfuse blood and look for another donor. [3] Self-control is carried out for the serum / plasma of the subject plus a 3-5% cell suspension in saline of the same subject. You can perform the technique on a plate, or better, with that tube in the same way as the crusade at RT. Normally red cells do not agglutinate when suspended in their serum / plasma. If you look at macroscopic agglutination This argues in favor of the presence of auto-antibodies or paraproteins (in this case will be observed under the microscope and the stacking of agglutination). A positive self-control requires a careful analysis of the agglutination test may be found in AB0 typing directly and indirectly through the repetition of tests At 37 ° tube. 
Determination AB0 indirect or "rebuttal"
It 's a test complementary and not alternative to direct determination. Its usefulness is mainly limited to the confirmation of the AB0 determined using commercial antisera allowing to interpret the possible, though infrequent, questionable results and to highlight the very rare false negatives and false positives of direct evidence .(....)
The counter is usually performed at the first determination group, while for the future control us is limited to the direct determination.
The indirect test for the presence of A and Anti B antibodies that are naturally present if the fees are absent antigens. To test should have known A and B red cells suspended in fresh saline to 3-4%. The red cell suspension may be stored in the refrigerator for several days. In practice this means having in the area known donors of group A and B available to frequent small blood samples.
Technique:
> mix 2 drops of serum or plasma from blood typing + 1 drop of red blood cells suspended in A or B notes to 3-4% in saline.
> read for agglutination
Even this test can be performed in test tubes (centrifugation and read out as above) or slide, slide, however, this survey is less reliable and requires more time and a closer reading eventually with the aid of a microscope at low magnification.
Expected Results:
° samples / plasma of group A and B red cells agglutinate 0 ° samples
/ plasma of group A red cells agglutinate B
° samples / plasma of group B red cells agglutinate A
° samples / group AB plasma did not agglutinate A or B red blood cells or red blood cells
If you are faced with a discrepancy between direct evidence and rebuttal, and that we are faced with the unexpected positive or negative, will need to make a series of additional tests to explain the reason for the discrepancy. In addition to a simple error in identification / execution of the test are different, but rare, situations that may explain unexpected results, by sub-groups AB0 rare (see the A2 subgroup [1] ) in the presence of autoantibodies cold [ 2] . In these cases to clarify the presence of indirect agglutination test is useful to the unexpected execution of a self [3] .
In general in the case of a recipient whose group AB0 found a discrepancy between direct and indirect evidence should be explained not only transfuse blood donor certainly 0.
Babies and up to 4 months of age do not yet have natural anti-A antibodies or anti-B, in these cases the proof is so often negative and its execution is merely confusing. Unexpected indirect negative test can also be observed in elderly or immunocompromised, in these cases, the absence or weakness of the reactivity is due to the low titer of regular.
A test of indirect confirmation of ABO compatibility of donor and recipient groups is represented by the cross-test at room temperature.
[1] Occasionally the plasma / serum A subgroup of A2 agglutinate red blood cells called type A1 is the most common. These clumps have no clinical significance, therefore the cells A1 can be safely transfused to patients with group A2 in spite of the apparent "incompatibility" [2] Presence of autoantibodies cold: in the case of agglutination of both cells ( A and B) which does not correspond to the direct determination, as expected, a group can be 0 in the presence of autoantibodies active at room temperature (antibodies or Fedde crioagglutinine). In these cases it is useful to add also a test of self-control in such cases will be positive, contrary to what is normally expected. If self-control is positive it is highly likely the presence of cold autoantibodies (that agglutinate at room temperature almost all the cells). These are generally not significant, that is not cause transfusion reactions. In case of positive self-repeating all the tests at 37 ° C by incubating cells and serum / plasma in a water bath. If the agglutination of self disappears (that is no longer active cold autoantibodies) can interpret the residual agglutination with red cells A or B or with those of the donor (during the test cross) as significant. If that persists in the agglutination test cross will be prudent not to transfuse blood and look for another donor. [3] Self-control is carried out for the serum / plasma of the subject plus a 3-5% cell suspension in saline of the same subject. You can perform the technique on a plate, or better, with that tube in the same way as the crusade at RT. Normally red cells do not agglutinate when suspended in their serum / plasma. If you look at macroscopic agglutination This argues in favor of the presence of auto-antibodies or paraproteins (in this case will be observed under the microscope and the stacking of agglutination). A positive self-control requires a careful analysis of the agglutination test may be found in AB0 typing directly and indirectly through the repetition of tests At 37 ° tube.
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AB0 Direct Determination
highlights the presence of A and B antigens on red blood cells by means of a typing agglutination reaction. It 's the basic method and simple, it can also be performed in emergency on a few drops of capillary blood obtained by puncture of the earlobe or digital .(....)
Reagents:
· Antiserum A Anti B and commercial [1] (keep refrigerated 2-6 ° C)
Standard:
· blood typing: anticoagulated (citrate or in a tube with EDTA is the preferred material) or fresh capillary (before clotting) or whole blood clot [2]
have 2 alternative methods:
· On white plate or slide:
> lay on the plate a drop of blood diluted + 1-2 drops of antiserum
> stir with a bar of plastic or glass cleaner
> tilting the plate to wait 30-60 seconds (or slide)
verify your agglutination.
• In test tube:
> put in a test tube one drop of cell suspension at 3-5% in saline [3] + 1-2 drops of antiserum
> centrifuge at low rpm (1000 xg approximately) for 30 seconds
> resuspend the bottom with light taps and rotation of the tube - tip and roll-to highlight the agglutination.
Interpretation:
§ §
presence of the agglutination = not = no agglutination antigen
Example: • Anti
A + = blood agglutination
• Anti B + blood = no agglutination
· Interpretation: A group
antisera on the market today are usually made of monoclonal antibodies, highly active, specific and stable even in optimal storage conditions. The reaction agglutination waiting with these commercial antisera is obvious to the naked eye and does not require the use of microscopic examination.
rarely possible false positives [4] , that is of real or apparent agglutination reactions that do not match the presence of the antigen, in this case A or B. The technique in test tubes with diluted red blood cells reduces false positives and questionable results.
E 'should perform regular quality control of the antisera with RBC notes (A, B, and 0) and most importantly it is essential to preserve the antisera in a working refrigerator and regularly check and verify their validity.
From a purely cultural point of view, we recall the existence rare subgroups of A and B that can give false-positive or questionable direct evidence.
[1] You can also use the antiserum anti-A, B, which agglutinate red blood cells both A and B. This serves as confirmation to the reactions observed with the 'anti-A el' anti-B. Today with the monoclonal antisera, more sensitive and specific than polyclonal human use in the past, the use of the antiserum anti-A, B is no longer indispensable. [2] E 'can also be used for the determination of coagulated blood group. After clot retraction test cells trapped the clot can be released and resuspended in serum breaking the branches of fibrin with a Pasteur pipette. [3] To prepare a cell suspension at 3-5% mix a drop of whole blood in 15 drops of saline solution or a drop of packed cells in 30 of saline solution. [4] False positive direct determination: this may be due to stacking of red blood cells (to be distinguished from agglutination), or you can see true agglutination caused by auto-antibodies in the blood plasma by typing, antibodies that affect temperature environment (called "cold"). In both cases, these factors present in blood plasma to be determined and are seen especially in the case of the technique on plate. In test-tube technique the plasma is very dilute, so the possibility of these false positives is also reduced in the test tube, the result is more clearly interpretable. Therefore, whenever possible, the test-tube technique is preferable.
In any case to exclude false-positive indirect evidence, particularly if the blood agglutinates with all the antisera available (in case of group AB + beware!), You should perform a self-control. 
Determination AB0 direct
highlights the presence of A and B antigens on red blood cells by means of a typing agglutination reaction. It 's the basic method and simple, it can also be performed in emergency on a few drops of capillary blood obtained by puncture of the earlobe or digital .(....)
Reagents:
· Antiserum A Anti B and commercial [1] (keep refrigerated 2-6 ° C)
Standard:
· blood typing: anticoagulated (citrate or in a tube with EDTA is the preferred material) or fresh capillary (before clotting) or whole blood clot [2]
have 2 alternative methods:
· On white plate or slide:
> lay on the plate a drop of blood diluted + 1-2 drops of antiserum
> stir with a bar of plastic or glass cleaner
> tilting the plate to wait 30-60 seconds (or slide)
verify your agglutination.
• In test tube:
> put in a test tube one drop of cell suspension at 3-5% in saline [3] + 1-2 drops of antiserum
> centrifuge at low rpm (1000 xg approximately) for 30 seconds
> resuspend the bottom with light taps and rotation of the tube - tip and roll-to highlight the agglutination.
Interpretation:
§ §
presence of the agglutination = not = no agglutination antigen
Example: • Anti
A + = blood agglutination
• Anti B + blood = no agglutination
· Interpretation: A group
antisera on the market today are usually made of monoclonal antibodies, highly active, specific and stable even in optimal storage conditions. The reaction agglutination waiting with these commercial antisera is obvious to the naked eye and does not require the use of microscopic examination.
rarely possible false positives [4] , that is of real or apparent agglutination reactions that do not match the presence of the antigen, in this case A or B. The technique in test tubes with diluted red blood cells reduces false positives and questionable results.
E 'should perform regular quality control of the antisera with RBC notes (A, B, and 0) and most importantly it is essential to preserve the antisera in a working refrigerator and regularly check and verify their validity.
From a purely cultural point of view, we recall the existence rare subgroups of A and B that can give false-positive or questionable direct evidence.
[1] You can also use the antiserum anti-A, B, which agglutinate red blood cells both A and B. This serves as confirmation to the reactions observed with the 'anti-A el' anti-B. Today with the monoclonal antisera, more sensitive and specific than polyclonal human use in the past, the use of the antiserum anti-A, B is no longer indispensable. [2] E 'can also be used for the determination of coagulated blood group. After clot retraction test cells trapped the clot can be released and resuspended in serum breaking the branches of fibrin with a Pasteur pipette. [3] To prepare a cell suspension at 3-5% mix a drop of whole blood in 15 drops of saline solution or a drop of packed cells in 30 of saline solution. [4] False positive direct determination: this may be due to stacking of red blood cells (to be distinguished from agglutination), or you can see true agglutination caused by auto-antibodies in the blood plasma by typing, antibodies that affect temperature environment (called "cold"). In both cases, these factors present in blood plasma to be determined and are seen especially in the case of the technique on plate. In test-tube technique the plasma is very dilute, so the possibility of these false positives is also reduced in the test tube, the result is more clearly interpretable. Therefore, whenever possible, the test-tube technique is preferable.
In any case to exclude false-positive indirect evidence, particularly if the blood agglutinates with all the antisera available (in case of group AB + beware!), You should perform a self-control.
Lorena Herrera Training
• 21 / 4 principles and practice in the HLA tissue typing laboratory (Dr. Laura UXA)
Tuesday, April 14, 2009
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