|Year : 2021 | Volume
| Issue : 1 | Page : 33-35
The screening and identification of atypical red cell antibodies by simultaneous LISS/coombs and NaCl/enzyme gel methods prior to blood transfusion
Purvee Agrawal, Arvind Bhake
Department of Pathology, Jawaharlal Nehru Medical College, Datta Meghe Institute of Medical Sciences (Deemed to be University), Wardha, Maharashtra, India
|Date of Submission||17-Apr-2020|
|Date of Decision||28-Dec-2020|
|Date of Acceptance||15-Feb-2021|
|Date of Web Publication||29-Jul-2021|
Dr. Purvee Agrawal
F-7 Meghe Heights, Jawaharlal Nehru Medical College Campus, Sawangi, Meghe, Wardha, Maharashtra
Source of Support: None, Conflict of Interest: None
Background: In this retrospective study, we measured the frequency of unexpected antibodies in the blood by Nacl/enzyme gel method or LISS/Coombs method and compared the result and frequency of identification in patients with a history of gestation or blood transfusion. Aims and Objectives: To detect unexpected/atypical red cell antibodies by simultaneous LISS/Coombs and NacL/enzyme gel methods prior to blood transfusion. Materials and Methods: Atypical antibody screening has done in all patients with the history of recurrent blood transfusion or history of gestation by commercially available red cell panel (ID–Dia panel, Diamed–ID Micro Typing System). Results: A total of 30 patients were screened for the presence of atypical antibody. Out of 30 patients, 1 (3.33%) patient showed the presence of alloantibody in the serum. The alloantibody identified was of anti-Lewis system. Conclusion: Screening for the presence of alloantibodies in patients is important to provide compatible blood and avoid adverse transfusion reactions.
Keywords: Alloantibody, atypical antibody, LISS
|How to cite this article:|
Agrawal P, Bhake A. The screening and identification of atypical red cell antibodies by simultaneous LISS/coombs and NaCl/enzyme gel methods prior to blood transfusion. J Datta Meghe Inst Med Sci Univ 2021;16:33-5
|How to cite this URL:|
Agrawal P, Bhake A. The screening and identification of atypical red cell antibodies by simultaneous LISS/coombs and NaCl/enzyme gel methods prior to blood transfusion. J Datta Meghe Inst Med Sci Univ [serial online] 2021 [cited 2021 Sep 16];16:33-5. Available from: http://www.journaldmims.com/text.asp?2021/16/1/33/322597
| Introduction|| |
Unexpected/atypical antibodies are sometime referred to as irregular antibodies because their existence and type are unknown before conducting an antibody screening test. This category of antibodies includes most blood type antibodies excluding ABO and some P blood type antibodies. Depending on the type of antibody, some develop naturally in patients but most unexpected antibodies are formed as part of the immune response after exposure to foreign antigens during pregnancy of transfusion.
Clinically significant unexpected/atypical antibodies are capable of causing hemolytic transfusion reactions secondary to accelerated destruction of a significant proportion of transfused red blood cells. Therefore, screening for unexpected antibodies should be part of all pretransfusion testing, with antibody identification in the event of a positive result. In the 1990s, the microcolumn gel technique was introduced for screening and identification of such unexpected antibodies. This method is not only easy to perform and economical of time but also easy to standardize and read, so it has become the most common technique in the blood bank laboratories of many countries.
The two principal techniques for unexpected antibody screening and identification are the indirect antiglobulin and enzyme methods. The most frequently used method is the indirect antiglobulin with gel (LISS/COOMBS), and the microcolumn assay technique using the LISS/Coombs gel test is the most popular for this purpose in Korea.,, However, the NaCl/enzyme method is used only for antibody identification.
Aim and objective
To detect unexpected/atypical red cell antibodies by simultaneous LISS/Coombs and NacL/enzyme gel methods prior to blood transfusion.
- To detect atypical/unexpected antibodies in a person receiving multiple transfusion by LISS/Coombs and NaCL/enzyme tests for the identification of atypical antibody
- To compare LISS/Coombs and NaCL/enzyme test methods for the detection of atypical antibody
- To determine the frequency of each antibody and its association with repeated transfusion or gestation.
Enzyme test, unexpected antibodies were further identified in 29 cases (anti-Lewis, 14; antirhesus, 13; and antiP1, 2). On the other hand, 9 cases (anti-M, 5; anti-Fyb, 3; and anti-N, 1) were identified by the LISS/Coombs test only. Of the unexpected antibodies found in patients without a previous history of transfusion or gestation, anti-Lewis (50.0%, 10/20) was the most common, while in patients with the history, antirhesus (48.1%, 26/54) was the most frequent.
The author concluded that NaCl/enzyme combined with LISS/Coombs gel test was useful for the identification of unexpected antibodies, and antibodies found in patients without a previous history of transfusion and gestation were clinically less relevant than those found in patients with the history.
| Materials and Methods|| |
Place of study: Blood bank, AVBRH Sawangi.
Duration of study: December 2017–December 2018.
Type of study: Observational (retrospective and prospective study).
Sample size: 30.
Technical Method: Atypical antibody detection and to identify the specificity of irregular erythrocyte antibody by commercially available red cell panel (ID–Dia panel, Diamed–ID Micro Typing System).
A 50 μl sample of 0.8% screening or identification cell reagent and 25 μl of patient serum were added to the microtube of each gel card. After 15 min incubation at 37°C , the card was centrifuged for 10 min and the reaction of agglutination was examined macroscopically on an illuminated view box.
All tests were carried out using the Diamed–ID Micro Typing System (DiaMed Ag, Cressier, Morat, Switzerland). For the LISS/Coombs screening method, the LISS/Coombs card and two test reagents ID-Diacell I-I (Diamed Ag) were used. For the NaCL/enzyme screening method, the NacL/enzyme identification test, the NaCL/enzyme card, and three test reagents Diacell I-II-III P (Papainized) (DiaMed Ag, ID) were used.
When unexpected antibodies were detected by either test, those antibodies were identified using both methods. For the LISS/Coombs identification test, the LISS/Coombs card and ID-panel test reagent (DiaMed Ag) were used. For the NaCL/enzyme identification test, the NaCL/enzyme card and the ID-Pane P test reagent (DiaMed Ag) were used.
Study material: Freshly drawn sample should be drawn in citrate, EDTA, or CPDA anticoagulant. Samples were drawn into the plain tubes (no anticoagulant).
Positive: Agglutinated cells forming a red cell line on the surface of the gel or agglutinate dispersed in the gel.
Negative: Compact button of cells on the bottom of the microtube.
Ethics and consent: No ethical issues risking the human life or animal is involved in the method carried out in the present study.
Statistical test: Chi-square test.
The Institutional Ethics Committee of DMIMSDU has approved the Research work proposed to be carried out at Jawaharlal Nehru Medical College, Sawangi(M), Wardha. Date: 17th Oct 2016 with Reference no DMIMS(DU)/IEC/2016-17/1120.
| Observation and Results|| |
A total 30 patients with a history of repeated blood transfusion including pregnant women with a history of multiple pregnancies were screened for the presence of atypical antibody. Out of 30 patients, 1 (3.33%) patient showed the presence of alloantibody in the serum. The alloantibody identified was of anti-Lewis system.
Among 30 patients, there were 13 males and 17 females; out of 17 females, 13 females showed a positive history of multiple pregnancy. Among 30 patients, a history of transfusion was present in 20 patients [Table 1].
| Discussion|| |
It is known, several difference in frequency and type of unexpected antibody are present in subjects who undergone repeated blood transfusion or pregnancy.
Previously, screening for unexpected antibodies was performed with an indirect antiglobulin test or an enzyme test using a conventional tube method. However, the recently introduced gel test has proved to be more sensitive and has many advantages.,, The use of LISS has increased the number of antibodies detected, and clinically important antibodies have been found in increasing numbers. In addition, the gel method is rapid, and interpretation of the results is easy. Therefore, the gel test gained widespread usage throughout the world, including Korea.
In this study, we compared the results of the LISS/Coombs and NaCl/enzyme tests using the gel method for screening and identifying unexpected antibodies and evaluated the clinical usefulness of simultaneous testing by two methods. Of the 30 patients' samples tested, 0% with the LISS/Coombs screening method and 3.33% NaCl/Enzyme. This result indicates that the NaCl/enzyme method is sensitive in detecting unexpected antibody, especially Rh antibodies.
Sol et al. also had similar results with unexpected antibodies were detected in 79 patients (1.43%). 39 (49.4%), 59 (74.7%), and 68 patients (86.1%) by the LISS/Coombs test, the NaCl/Enzyme test, and the two tests combined, respectively. With the addition of the NaCl/enzyme test, unexpected antibodies were further identified in 29 cases.,,
The Lewis antibody, which was the most frequently identified antibody in our study, is an IgM antibody which cannot pass through the placenta and so is unrelated to hemolytic disorders in newborns. This antibody, which is detected through direct agglutination, can occasionally cause hemolytic complications associated with transfusion and is frequently found in expectant mothers with no great clinical significance. Ko et al. also have similar findings with our study. However, Han et al. determined that anti-Le a, which is a type of Lewis antibody, causes delayed hemolytic reactions. The facts that anti-Le a antibodies are mostly formed not from immune response but spontaneously and that the patient in their case had no history of pregnancy or transfusion were cited as the reasons for their conclusion.,,
| Conclusion|| |
Screening of patient with repeated blood transfusion and multiple pregnancies is important and detection of alloantibody in donor blood is important to provide compatible blood to the receivers and to avoid adverse transfusion reaction in as patient with unexpected blood antibody may have increased risk of delayed transfusion.
Financial support and sponsorship
Conflicts of interest
There are no conflicts of interest.
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