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 Table of Contents  
ORIGINAL ARTICLE
Year : 2021  |  Volume : 16  |  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


Department of Pathology, Jawaharlal Nehru Medical College, Datta Meghe Institute of Medical Sciences (Deemed to be University), Wardha, Maharashtra, India

Date of Submission17-Apr-2020
Date of Decision28-Dec-2020
Date of Acceptance15-Feb-2021
Date of Web Publication29-Jul-2021

Correspondence Address:
Dr. Purvee Agrawal
F-7 Meghe Heights, Jawaharlal Nehru Medical College Campus, Sawangi, Meghe, Wardha, Maharashtra
India
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Source of Support: None, Conflict of Interest: None


DOI: 10.4103/1319-4534.322597

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  Abstract 


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 Top


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.[1]

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.[2] 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.[3] 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.[4]

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.[5],[6],[7] However, the NaCl/enzyme method is used only for antibody identification.

Aim and objective

Aim

To detect unexpected/atypical red cell antibodies by simultaneous LISS/Coombs and NacL/enzyme gel methods prior to blood transfusion.

Objectives

  1. To detect atypical/unexpected antibodies in a person receiving multiple transfusion by LISS/Coombs and NaCL/enzyme tests for the identification of atypical antibody
  2. To compare LISS/Coombs and NaCL/enzyme test methods for the detection of atypical antibody
  3. 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 Top


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).

Interpretation

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.

Ethical clearance

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 Top


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].
Table 1: Characteristics of patients (n=30)

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  Discussion Top


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.[5],[10],[11] 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.[5] 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.[8],[9],[10]

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.[12] also have similar findings with our study. However, Han et al.[11] 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.[13],[14],[15]


  Conclusion Top


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

Nil.

Conflicts of interest

There are no conflicts of interest.



 
  References Top

1.
Han KS, Park MH, Kim SI. Transfusion Medicine. 3rd ed.. Seoul, Korea: Medical Publisher; 2006. p. 153-287.  Back to cited text no. 1
    
2.
Chapman JF, Elliott C, Knowles SM, Milkins CE, Poole GD, Working Party of the British Committee for Standards in Haematology Blood Transfusion Task Force. Guidelines for compatibility procedures in blood transfusion laboratories. Transfus Med 2004;14:59-73.  Back to cited text no. 2
    
3.
Lapieere Y, Rigal D, Adam J, Josef D, Meyer F, Greber S, et al. The gel test a new way to detect red cell antigen-Antibody reactions. Transfusion 1990:30:109-13.  Back to cited text no. 3
    
4.
de Castilho LM, Pellegrino J Jr., Bechelli AP, Le Pennec PY, Mendes NF. Evaluation of recent techniques for detection of red blood cell antibodies in sera of reference samples, patients, pregnant women, and blood donors. J Clin Lab Anal 1996;10:250-6.  Back to cited text no. 4
    
5.
Lee WH, Kim SY, Kim HO. The incidence of unexpected antibodies in transfusion candidates. Korean J Blood Transfuse 2000;11:99-103.  Back to cited text no. 5
    
6.
Cho YK, Kim DS, Lee HS, Choi SI. The frequency and distribution of unexpected antibodies in surgical patients at Chonbuk National University Hospital. Korean J Lab Med 2004;24:67-71.  Back to cited text no. 6
    
7.
Jung TK, Lee NY, Bae HG, Kwon EH, Park SH, Suh JS. Unexpected antibody positivity with the use of the LISS/Coombs gel test. Korean j Clinpathol 2001;21:422-58.  Back to cited text no. 7
    
8.
Garg N, Sharma T, Singh B. Prevalence of irregular red blood cell antibodies among healthy blood donors in Delhi population. Transfus Apher Sci 2014;50:415-7.  Back to cited text no. 8
    
9.
Bromilow IM, Adams KE, Hope J, Eggington JA, Duguid JK. Evaluation of the ID-gel test for antibody screening and identification. Transfus Med 1991;1:159-61.  Back to cited text no. 9
    
10.
Nathalang O, Chuansumrit A, Prayoonwiwat W, Siripoonya P, Sriphaisal T. Comparison between the conventional tube technique and the gel technique in direct antiglobulin tests. Vox Sang 1997;72:169-71.  Back to cited text no. 10
    
11.
Han KS, Cho HI, Kim SI. A study on the hemolytic transfusion reactions due to irregular antibodies. Korean J Hematol 1989;24:2733.  Back to cited text no. 11
    
12.
Ko KH, Yoo BH, Kim KM, Lee WY, Yon JH, Hong KH, et al. Frequency of unexpected antibody and consideration during transfusion. Korean J Anesthesiol 2012;62:412-7.  Back to cited text no. 12
    
13.
Chandi DH, Patil P, Damke S, Basak S, Ashok R. Bacteriologic Antibiography Outline of Isolates from Blood Culture at Tertiary Center. J Pure Appl Microbiol 2020;14:2801-6. Available from: https://doi.org/10.22207/JPAM.14.4.55. [Last accessed on 2020 Jun 18].  Back to cited text no. 13
    
14.
Mohammad S, Bhute A, Acharya N, Acharya S. Moschcowitz Syndrome or Thrombotic Thrombocytopenic Purpura and Antiphospholipid Antibody Syndrome as a Rare Cause of Thrombocytopenia in Pregnancy Mimicking Hemolysis, Elevated Liver Enzymes, and Low Platelets Syndrome in a Patient with Bad Obstetric History: A Diagnostic Dilemma. J SAFOG 2020;12:250-3. Available from: https://doi.org/10.5005/jp-journals-10006-1791. [Last accessed on 2020 Jun 18].  Back to cited text no. 14
    
15.
Yadav N, Madke B, Kar S, Gangane N. Netherton Syndrome: An Atypical Presentation. Cutis 2019;103: E27-9.  Back to cited text no. 15
    



 
 
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