Multi-transfused Thalassemia Patients Research Articles

Through the Eyes of the Recipient: Navigating Transfusion Services Amidst COVID-19 in Multi-Transfused Thalassaemic Patients

Through the Eyes of the Recipient: Navigating Transfusion Services Amidst COVID-19 in Multi-Transfused Thalassaemic Patients

Alloimmunization and autoimmunization among multitransfused thalassemia and sickle cell disease patients

Blood transfusion is the mainstay for management of hemoglobinopathies, including thalassemia and sickle cell disease (SCD). Apart from other transfusion related adverse effects, immunization is a significant complication, particularly in multitransfused recipients. Alloimmunization rates vary widely, both in thalassemia and SCD patients. Alloimmunization complicates the management in form of hemolytic transfusion reactions, difficulty in finding compatible units, delays in transfusion and increasing costs. Exact pathogenesis and prevention modality have not been elucidated. Apart from antigenic differences, immunomodulatory effects and inflammation also play a role in pathogenesis. Ethnic heterogeneity between donors and recipients predisposes to higher alloimmunization.Transfusion management of hemoglobinopathies across the globe is fragmented, with different levels of care. Guidelines propose red cell antigen profiling, transfusion of leucoreduced red cells, preferably matched for Rh and Kell antigens, over and above ABO-D matching. For alloimmunized patients, corresponding antibody negative red cells need to be transfused. However, patients from different backgrounds are variably transfused with whole blood/red cells, which may or may not be leukoreduced. Prophylactic antigen matching reduces the risk of alloimmunization, however, has limitations. RH variants may not be picked up by phenotyping alone. Moreover, the approach of prophylactic antigen matching is expensive, labour intensive, time consuming and may cause delays in transfusions. There is limited, low quality evidence on best practices for transfusion management of SCD and thalassemia. The review analyses the magnitude of problem, factors contributing and modalities for prevention and management of immunization.

Prevalence of Kell Blood Group Antigens among Blood Donors & Impact of its Alloimmunization in Multi-transfused Thalassemia & Sickle Cell Disease Patients with Recommendation of Transfusion Protocol—Need of the Hour

Background: The aim of the study was to analyze the prevalence of Kell antigen and its correlation to major blood groups, ABO & Rh system in Eastern India. There was simultaneous retrospective analysis of Kell alloimmunization to find out the implication and recommendation of transfusion protocols in multi-transfused thalassemia and sickle cell patients. Methods: The study was a prospective observational conducted on 3000 donors for KELL and ABO grouping. Retrospective analysis was made to identify common alloantibodies in multi-transfused patients. Results: The overall prevalence of Kell antigen was 2.6% (80) in 3000 donors. Among male, it was highly prevalent i.e. 2.77% and in females 0.65%. Kell antigen was highly prevalent among AB donors, i.e. 5.1%. It was 2.5% in A, 2.9% in B, 1.9% in Blood Donors. Kell prevalence was high in Rh D positive donors, i.e. 2.72% and was 1.72% among Rh D negative donors. Anti-K was the 3rd most common alloantibody detected in 638 cases of multi-transfused thalassemia and sickle cell (SCD) patients (9.25%). Anti-E (42.6%)was most common entity followed by anti-c (24.0%). Conclusion: The higher incidence of Kell prevalence in AB & Rh D Positive Blood groups and also in male persons indicate that there should be a donor database and knowledge of red cell antigen prevalence in a population. This will help blood centers in providing antigen negative compatible blood units to patients with corresponding alloantibodies. Hemolytic transfusion reactions due to Kell alloimmunization are of a significant severity. Prevalence of Kell alloantibody is high among multi-transfused patients and is next to anti E & anti c. Kell sensitized mothers may also cause serious consequences like hemolytic disease of fetus and newborn. Therefore, it is suggested that extended phenotyping including Kell blood group antigen should be implemented in cases of multi-transfused patients. Keywords: Blood donors, Extended phenotyping, Kell, Multitransfused thalassemia patients, Multitransfused sickle cell disease patients

Evaluation of diastolic dysfunction in multi-transfused thalassemia patients

Background: An important complication of thalassemia is iron deposition in the cardiac tissue resulting in degeneration, fibrosis and dysfunction. Cardiac disease is the primary cause of death. Aims and objectives were to detect early cardiac involvement in patients with transfusion dependent thalassemia by conventional doppler echocardiography and tissue doppler imaging while they are still asymptomatic. Methods: 150 patients in the age group of 2-18 years who were receiving regular and frequent blood transfusions were enrolled in this study. Institutional ethical clearance was taken. After informed consent, 2D echocardiography was performed on all the patients. Previous blood reports reviewed. Various parameters like mitral flow pattern, LVEF and Tricuspid regurgitant jets were taken into account to diagnose cardiac dysfunction. Results analysed by standard statistical tests. Results: Out of 150 patients, 58% were males and 42% were female patients. The 13 (8.6%) patients were diagnosed as having diastolic dysfunction. Out of 150, majority (86.6%) were below the age of 15 years, 81.33% children were diagnosed as having thalassemia before the age of 2 years, 97% patients were on regular iron chelation therapy, 32% patients had pre-transfusion Hb <7 gm/dl, 78.66% patients had S. ferritin levels >2500 ng/ml and more number of males (7.33%) have undergone splenectomy than females. Conclusions: Diastolic dysfunction on Doppler echocardiography is an early sign of myocardial dysfunction in patients with beta thalassemia.

A comparison of serological phenotyping and molecular genotyping for Kell, Kidd, and Duffy antigens in multi-transfused thalassemia patients.

In multi-transfused thalassemia patients, serological phenotyping fails to test patient's actual blood group antigen profile due to the presence of donor red blood cell (RBC) in the circulation. This limitation of serological tests can be overcome by genotype determination using the polymerase chain reaction (PCR)-based methods. The aim of this study is to compare the serological phenotyping of Kell, Kidd, and Duffy blood group systems with molecular genotyping in the normal blood donors and multi-transfused thalassaemia patients. Blood samples from 100 normal blood donors and 50 thalassemia patients were tested using standard serological techniques and PCR-based methods for Kell (K/k), Kidd (Jka/Jkb), and Duffy (Fya/Fyb) blood group systems. The results were compared for concordance. Genotyping and phenotyping results were 100% concordant for normal blood donors whereas those for thalassemia patients showed 24% discordance. The frequency of alloimmunization in thalassemia patients was 8%. The results of genotyping were used to provide Kell, Kidd, and Duffy matched blood for transfusion therapy to thalassemia patients. The actual antigen profile in multitransfused thalassaemia patients can be reliably determined using genotyping. This would benefit in providing better antigen matched transfusion therapy to such patients hence reducing the rate of alloimmunization.

Prevalence of Transfusion-Transmitted Infections in Multi-Transfused Thalassemia and Sickle Cell Disease Patients: A Single Center Experience from Southern Odisha, India

ABSTRACT Background: Repeated transfusion in sickle cell disease (SCD) and transfusion-dependent thalassemia (TDT) patients lead to transfusion-transmitted infections (TTIs). The prevalence, as well as various risk factors for TTIs in these patients, is found to be inconsistent. This study aimed to observe the prevalence, types of infections as well as risk factors for TTIs in a tertiary health care center. Materials and Methods: This observational study was undertaken on both SCD and TDT patients admitted to the hospital for a transfusion. In all patients, the enzyme-linked immunosorbent assay-based test kit was used for the diagnosis of hepatitis C virus (HCV), hepatitis B virus (HBV), and human immunodeficiency virus 1 and 2 (HIV 1 and 2). However, a rapid test kit was used for the diagnosis of malaria and syphilis. Detailed information on age of the patients, duration of transfusion, and number of transfusion as risk factors were recorded. The association of factors with various infections was analyzed statistically. Results: A total of 153 patients were recruited including 49 SCD and 104 TDT patients. Of 153 patients, 25 patients were found to have infections including HCV in 22 (14.38%) patients, HBV in 2 (1.31%) patients, and HIV in 2 patients (1 patient was positive for both HCV and HBV). Out of 25 TTIs patients, 7 patients had SCD and 18 patients had TDT. On the association, only an increased number of transfusion per year was found to be significantly associated with TTIs (P = 0.008) in SCD patients. However, all the risk factors were found to be associated with TTIs except gender in TDT patients. Conclusion: The occurrence of TTIs was found to be associated with older patients, increased number of transfusion per year, and large duration of transfusion. The increasing trend in the TTIs in these patients calls for a shift toward preventive and comprehensive care to reduce the health burden.

Alloimmunization and Autoimmunization in Multi-Transfused Thalassemic Patients: A Single Center Study

Aim: To determine frequency of alloimmunization and autoimmunization among the patient of thalassemia with history of multiple blood transfusions. Methods: This study was conducted by consecutive sampling in six months duration from September 2020 to August 2021 at the department of Hematology and blood Transfusion Medicine, of University of Child Health Sciences, The Children's Hospital Lahore. Any patient with a positive Direct antiglobulin test (DAT) was labeled as auto-immunized and a positive indirect antiglobulin test (IAT) was labeled as alloimmunized. Results: Total 90 cases were enrolled with 62% male and 38% females with a mean age 6.04 + 3.3 years (range of age: 7 months to 14 years). Antibody screening and auto-antibodies were positive in 4.4% (n= 4) cases each, and allo-antibodies in 6.7% (n=6) cases (anti-E in 2, anti-C in 2, anti-K and anti-e in 1 each). Among all these patients of beta thalassemia, spleen was enlarged in 71% cases; around half of these were <5 years old. Around 96% had first transfusion <2 years. The frequency of patients positive on DAT, IAT and on anti-body screening was significantly related to the frequency of blood transfusion Practical Implications: Autoantibodies produce a positive direct antiglobulin test (DAT), resulting in hyperhemolysis of red cells in thalassemic patients, which exacerbates the existing alloantibodies factor in children who have received several blood transfusions. The gap between transfusions will be shortened. Routinely, greater emphasis is placed on the identification of allo-antibodies, but auto-antibodies should also be evaluated so that this phenomena may be better understood and handled. This study was conducted to examine the link between a greater transfusion demand and the existence of auto or allo antibodies in Thalassemia patients. I Conclusion: We found Alloimmunization and autoimmunization together not very uncommon in transfusion dependent patients of Thalassemia. The frequency of transfusions is affected in co-existing states. Extended matching, as well as early diagnosis and control of Auto and allo-antibodies both can improve the efficacy of blood transfusion. Keywords: Autoantibodies, IAT, DAT, Coomb’s test, Thalassemia, Allo-immunization

Prevalence and Molecular Evaluation of Hepatitis C Virus Infection among Multi-transfused Thalassemia Patients in South of Iran.

ObjectivesInformation regarding the magnitude of hepatitis C virus (HCV) infection among thalassemia patients is of great importance for health care providers to assess blood safety and improve the quality of screening systems. Therefore, this study evaluated the prevalence, risk factors, and genotypic pattern of HCV infection among β-thalassemia patients in South Iran.MethodsThis descriptive-analytical cross-sectional study was conducted from March to June 2019. All patients with β-thalassemia major from Borazjan, Bushehr, Delvar, Kangan, and Ahram cities participated in the study and attended the transfusion center of the Bushehr University of Medical Sciences located in southern Iran. Serum samples were tested for the presence of anti-HCV antibodies by an enzyme-linked immunosorbent assay. The seropositive serum samples were tested for detection of HCV viremia and genotypes by semi-nested reverse transcriptase-polymerase chain reaction and sequencing.ResultsOf 125 thalassemia patients, 22 (17.6%) were positive for anti-HCV antibodies and two (1.6%) had HCV viremia with genotype 3a. HCV seroprevalence increased with age, so anti-HCV seropositive thalassemia patients had significantly higher mean age than anti-HCV seronegative patients. HCV seroprevalence was higher among female patients, residents of Kangan, patients with blood transfusion every two weeks, Fars patients, and thalassemia patients with alanine aminotransferase levels of < 20 IU/L and aspartate aminotransferase levels of > 80 IU/L. Nevertheless, anti-HCV seroprevalence among thalassemia patients was not statistically associated with these variables.ConclusionsThese results indicate ongoing HCV incidence among the thalassemia population in this region. Transfusion of HCV-seronegative viremic blood units donated during the infectious window period contributes to HCV infection in thalassemia patients. These findings highlight the need to include sensitive molecular assays in the screening process of donated blood for HCV infection in Iran.

SEROPREVALENCE OF HEPATITIS B VIRUS AMONG MULTI-TRANSFUSED BETA THALASSEMIA PATIENTS ATTENDING AT THE YEMENI SOCIETY FOR THALASSEMIA - SANA'A, YEMEN

Regular blood transfusion is one of the most important and emergency treatment methods for thalassemia patients. This category of patients is considered sensitive to infection because their continued survival is linked to blood transfusion, and therefore the possibility of infection is great, especially in light of the war and the deterioration of health services in Yemen. Among this infection is the hepatitis B virus, which causes serious complications such as acute hepatitis, liver cirrhosis and hepatocellular carcinoma (HCC). The aim of the study was done to determine incidence of hepatitis B virus and risk factors among Multi-Transfused Beta Thalassemia Patients attending at The Yemeni Society for Thalassemia and Genetic Blood Disorder (YSTGBD) Sana'a –Yemen. This study was conducted during the period from January 2021 to December 2021, during which 200 Blood samples were collected from beta thalassemia patients attending at (YSTGBD). All samples were examined using the ECLIA technique, during which the basic information was collected through special questionnaires for this purpose. The rate of incidence by HBsAg in all study samples was determined to be 7(3.5%). The study showed that there was a statistically significant relationship between infection with the hepatitis B virus and the age group of more than 10 years age (P = 0.042), in addition to that number of blood transfusions received by HBsAg positive was significantly higher than that of HBsAg negative thalassemia patients (P = 0.022).

Burden estimation of hepatitis B &amp; C in thalassemia patients in MGM Medical College and Hospital, Jamshedpur

Introduction: Thalassemia encompasses a group of genetic blood disorders, in which there is altered or inadequate haemoglobin synthesis and requires a repeated blood transfusion. Due to repeated blood transfusion, these patients are vulnerable to HCV, HBV, and HIV like blood transfusion transmitted diseases. Aims and Objective: To determine the prevalence of HCV and HBV infections in multitransfused thalassemia patients in a tertiary care hospital. Material And Methods: Cross sectional and observational studywas done on a total of 110 beta thalassemia patients aged between 2-25years &amp;were categorised into four groups (2-7 years, group 1; 8-13 years, group 2; 14-19 years, group 3 and 20-25 years, group 4).For testing, 5 ml Blood sample was collected, Serum separatedand subjected to testing for HBsAg, and antibody to Hepatitis C, by ELISA method. Screening was done by immunochromatography test (point of care test). Results: A total of 110 patients of beta thalassemia major were included in our study. Out of this, 68(62%) were males and 42(38%) were females.

High Frequency of Post-Transfusion Microchimerism Among Multi-Transfused Beta-Thalassemic Patients.

BackgroundTransfusion-associated microchimerism implies the presence of allogeneic hematopoietic cells in an individual, following the transfusion of a blood product. It is a transfusion-related adverse effect/long-term consequence, which has not been well-investigated among regularly transfused patients with thalassemia.Patients and MethodsWe investigated 64 regularly transfused, homozygous β-thalassemic patients and 21 never-transfused healthy volunteer blood donors (controls) for the presence of microchimerism in their sera, using real-time PCR targeting circulating allogeneic, both, Human Leukocyte Antigen-DR isotype (HLA-DR) and non-HLA alleles. The investigation was longitudinally repeated in patient subsets for more than 2 years. Results were correlated with clinical and laboratory parameters, peripheral blood lymphocyte immunophenotype, blood storage time, and donor's gender to identify potential contributing factors for microchimerism generation.ResultsOverall, microchimerism was detected in 52 of the 64 patients (81.2%) and in 6 of the 21 controls (28.5%, p = 0.0001). Forty-four patients (68.7%) exhibited long-term microchimerism (persisted for more than 6 months), confirmed at all time-points investigated. Microchimerism was more frequent among elderly, women, splenectomized and more heavily transfused patients, and among those who exhibit higher serum ferritin levels. In these patients, a distinct descending pattern of CD16dim+CD56dim+ natural killer (NK)-cells (p < 0.001) and an ascending pattern of CD4+CD25brightCD127– regulatory T-cells (p = 0.022) for increasing allelic burden were noticed, suggesting the establishment of recipient immune tolerance against the donor-derived chimeric alleles. Both splenectomized and non-splenectomized thalassemic patients exhibited the same trend. The storage time of transfused blood products and donor/gender mismatch had no impact on the development of microchimerism.Discussion-Conclusive RemarksTransfusion-associated microchimerism appears to be a very common complication among multi-transfused thalassemic patients. The potential clinical consequences of this phenomenon remain as yet unclear. Immune tolerance attributed to disease itself and to repeated transfusions might at least in part explain its appearance.

Cw Alloimmunization in Multitransfused Thalassemic Patients of North India

Background and Objectives: The C Willis or Cw antigen is a low-incidence antigen of Rh system. The antibody against the Cw antigen (anti-Cw) is an IgG antibody which may occur naturally or may be immune in nature. The identification of Cw antibody is important since it has the potential to cause hemolytic disease of the newborn as well as hemolytic transfusion reaction. This study was conducted with the aim of determining the prevalence of Cw antibody in multitransfused thalassemic patients enrolled in a Regional Blood Transfusion Center (RBTC) of North India. Methods: A retrospective descriptive observational study was conducted at the Department of Immunohematology and Blood Transfusion, LHMC and Associated Hospitals. All transfusion-dependent thalassemic (TDT) patients and non-TDT (NTDT) patients enrolled in the RBTC of the hospital till December 2018 were included in the study. Antibody screening was performed in all recipients before each transfusion. The prevalence of anti-Cw was estimated. Results: A total of 567 thalassemic patients (including TDT and NTDT) were registered in RBTC, LHMC till December 2018. On pretransfusion antibody screening and identification, 3 out of 567 thalassemic patients were found to have alloimmunization against Cw antigen. The prevalence of anti-Cw in multitransfused thalassemics was 0.53%. Conclusion: The prevalence of anti-Cw is variable in different populations and it is not a very commonly reported antibody in patients with thalassemia. One should be aware of the approach to transfusion in thalassemic patients who develop Cw alloimmunization.

Red Blood Cell Immunization and Contributing Factors in 685 Thalassemia Patients.

Background: An analysis of red blood cell alloimmunization in patients with thalassemia can help to devise specific strategies to decrease the alloimmunization rate. This study explored the frequency and specificity of alloantibodies and autoantibodies against red blood cell (RBC) antigens in patients with thalassemia referring to the Iranian Blood Transfusion Organization (IBTO) Immunohematology Reference Laboratory (IRL) in Tehran. Materials and Methods: This study first examined the laboratory records of 23,113 patients suffering from different diseases referring to IBTO’s IRL for pretransfusion testing in the 2008-2015 period. ABO and Rh(D) typing and antibody screening tests were performed for all 23,113 patient records and 685 (2.97%) beta-thalassemia patients with positive pre-transfusion test results (antibody screening and/or DAT) were selected for further investigation. Results: The antibody screening test was positive in 640 out of 685 thalassemic patients (93.4%). DAT was performed for 529 patients, 226 (33%) of which showed positive results. Meanwhile, 161 out of 685 beta-thalassemia patients (23.5%) had positive auto control test results, reflecting the possible presence of allo- and/or autoantibodies. The most common antigen-specific alloantibodies were directed against K and E RBC antigens with a frequency of 25% (Anti-K) and 11.91% (Anti-E), respectively. The development of two antibodies (double antibodies) in one patient was observed in 80 individuals (11.46%). Conclusion: Age, gender, history of pregnancy, and splenectomy were not contributing factors to the antibody presence in the patient population under study. Extended red blood cell phenotyping should be considered as an essential procedure for expected multi-transfused thalassemia patients before blood transfusion. Considering the high frequency of anti-K and anti-E observed in this study, it is recommended that thalassemia patients in Iran are tested through phenotyping of RBC units for K and E antigens before transfusion.

The Prevalence of Viral Hepatitis B and C among Multitransfused Thalassaemic Patients in Azadi Teaching Hospital in Kirkuk City

Background: Patients with thalassemia major are at high risk of hepatitis C and B due to the blood transfusion from donors infected by HCV and HBV. Hepatitis C virus (HCV) is the major cause of post-transfusion hepatitis infection (PTH). Objective: The aim of this study is to detect the prevalence of anti-HCV antibodies and HBs Ag and risk factors in multitransfused thalassemic patients in Kirkuk city to establish better preventive strategies. Patients and Methods: This study is conducted in Kirkuk city from Nov 2012 to March 2013 for HCV and HBV infection. A structured interview questionnaire is developed by the trained researcher to collect the demographic and risk factors. Statistical analysis was done by Chi-square test. Results: The prevalence of HCV is estimated to be (17.8%) and this is increased with increasing the requirement for blood transfusion in splenectomized patients while only (2.0%) is positive for HBV infection. Conclusion: The study shows a higher prevalence of HCV infection among multitransfused thalassemic patients with increasing requirement for blood transfusion in splenectomized patients.

Molecular and genetic characterization of hepatitis B virus among multitransfused thalassaemia patients in Islamabad, Pakistan.

Background:Hepatitis B virus (HBV) is the aetiological agent of transfusion-transmitted hepatitis globally. Beta thalassaemia major individuals are at greater risk of contracting HBV infection due to multiple blood transfusions required for the medical management of these patients. Based on HBV genetic variability, it is divided into 10 genotypes. The determination of HBV genotypes has significant implications for clinical management and treatment regimens.Aim:This study was performed to assess the HBV epidemiology and circulating genotypes in multi-transfused β-thalassemia major patients with the aim to be considered while formulating the treatment pattern taking into account particular needs of thalassaemia patients.Materials and Methods:This study was performed from September 2018 to June 2019, at the Department of Pathology and Transfusion Medicine, Shaheed Zulfiqar Ali Bhutto (SZAB) Medical University, Islamabad. A total of 2,260 thalassaemia patients were enrolled in the study. The study was endorsed by the Ethics Committee of the SZAB Medical University, Islamabad. The samples were serologically screened for HBsAg on the LIAISON® XL Murex HBsAg Quant assay (DiaSorin S.p.A., Italy) a chemiluminescence based immunoassay (CLIA). HBV quantitative PCR kit was used to measure the HBV DNA in serum samples. The HBV genotypes were determined using universal primers targeting the P1 and S1 region amplification.Results:Of 2,260 thalassaemia patients, 64.6% were males while 35.4% were females. The HBsAg was identified in 98 individuals (4.33%). The PCR analysis was done for these 98 patients and in this cohort, genotype D was 59.18% (n = 58), genotype A was 21.42% (n = 21) while genotype C was 19.38% (n = 19).Conclusion:The determination of HBV genotypes in the multi-transfused patients is key to the effective management of chronic HBV patients as the severity and course of the disease is dependent on a specific type of genotypes. Quality assured screening of donated blood will prevent the incidence of HBV in thalassaemia patients.

Genotyping analysis of the MNS blood group system of thalassemia patients with alloantibodies in Iran

BackgroundSerological methods are unreliable for accurate determination of blood group antigens in multi-transfused thalassemia patients. The MNS blood group system has five high-frequency antigens. Many studies demonstrated that some antibodies including anti-S, anti-s, and anti-U may cause acute and delayed transfusion reactions and hemolytic disease of the fetus and newborn. This study aimed to determine the genotype of the MNS blood group in thalassemia patients with alloantibodies by molecular methods. Material and methodsIn this study, 104 blood samples from thalassemia patients were collected. The blood group phenotype for M, N, S and s antigens was determined by the tube hemagglutination method. MNS blood group genotyping was performed using PCR-SSP and DNA Sequencing methods. ResultsAll patients were genotyped with a total of 6 pairs of primers. Discrepancies between genotype and phenotype were observed in 22 patients with S/s alleles and 2 patients with M/N alleles, however, there was full accordance between the results of SSP-PCR and DNA sequencing. The frequency of MNS blood group alleles was determined as follows: 25 % MNSs, 23 % MNss, 21 % MMSs, 9% MMSS, 9% MMss, 8% NNss, 2%MNSS, and NNSS, NNSs, MM genotypes at 1% each. ConclusionIn conclusion, molecular genotyping is more reliable than serological methods in multiple transfusion patients and can lead to a more compatible blood unit for transfusion in these patients.

Genomic analyses of KEL alleles in alloimmunized thalassemia patients from Iran

ObjectivesSerological methods are unreliable for red blood cells (RBCs) antigen typing in multi-transfused thalassemia patients due to the presence of donor RBCs in the recipient’s circulation and interfering antibodies. Kell blood group system is important in transfusion medicine and Kell antibodies have shown as the most prevalent antibodies in thalassemia patients. We intended to determine the genotype of Kell antigens among Iranian alloimmunized thalassemia patients using molecular methods and compare the results with serological phenotyping. MethodsTwo hundred thalassemia patients participated in this study. Blood group phenotype was performed by the serological method, while the genotype was determined for KEL*01, KEL*02, KEL*03, and KEL*04 alleles using PCR-Sequence Specific Primer (PCR-SSP) method. The genotypes of patients with incompatibility between phenotype and genotype were re-evaluated by Restriction Fragment Length Polymorphism-PCR (RFLP-PCR) and confirmed by DNA sequencing in all cases. ResultsTen patients were found with discrepancies between genotype and phenotype; however, there was a complete agreement between the results of SSP-PCR, RFLP-PCR, and DNA sequencing. Six discrepancies were found in the KEL*01/KEL*02 allele when serologically phenotyped as K-k+. One patient with K-k- and three patients with Kpa-Kpb + phenotype were identified as KEL*01/KEL*02 and KEL*03/KEL*04, respectively. ConclusionIt can be concluded that molecular genotyping is more reliable compared with the serological method, especially in the patients who have received multiple transfusions. Therefore, using a combination of these techniques can lead to a better matched transfusion in these patients.

Clinical and Biochemical Assessment of Liver Function Test and Its Correlation with Serum Ferritin Levels in Transfusion-dependent Thalassemia Patients.

AimsThe aim of our study was to correlate liver function tests with serum ferritin levels in multi-transfused thalassemia patients.Methods This was a descriptive cross-sectional study conducted in the department of hematology, Khyber Medical University, from January 2018 to December 2018. Thalassemia patients of either sex dependent on transfusion ≥ 1 year and having a confirmatory report of the disease were included in our study. The nonprobability convenience sampling technique was used. The Pearson correlation coefficient was applied to observe the correlation between serum ferritin level and liver function tests. A p-value of ≤0.05 was considered statistically significant. SPSS version 23 (SPSS Inc., Chicago, Illinois) was used for data analysis.ResultsA total of 138 subjects of age range 2-23 years, with a mean age of 12.08 ± 6.02 years, were included in our study. The mean serum ferritin of patients in our study was 3278.64 ng/ml with the lowest of 285.2 ng/mL and the highest of 10940.2 ng/ml. With the increase in serum ferritin levels, a rapid increase in alanine aminotransferase (ALT), aspartate aminotransferase (AST), and alkaline phosphatase (ALP) levels was seen. When serum ferritin levels were correlated with total bilirubin level, the bilirubin level remains static with a further increase in serum ferritin levels.ConclusionIt was deduced that iron deposition is the ultimate reason for increased liver enzymes. There was a positive correlation between serum ferritin and ALT, AST, and ALP while a weak connection was found between serum ferritin and bilirubin levels.

Detection of red cell alloantibodies in thalassaemia patients

Background: β-thalassaemia patients receive regular blood transfusion to thrive. Due to antigen disparity between the blood donors and these patients they develop red cell alloantibodies due to alloimmunization. The objective of this study is to predict the frequency of red cell alloimmunization amongst β-thalassaemia major patients receiving regular blood transfusion.Methods: This study including 106 patients with β-thalassaemia was conducted in the department of Transfusion Medicine, S. C. B. Medical College, Cuttack for a period of 12 months. Alloantibodies to different red cell blood group antigens in multi-transfused thalassaemia patients were detected using the glass bead technology for blood group serology in the present study.Results: Out of 106 β-thalassaemia major patients included in the study, 7.5% of patients developed alloantibodies, all being clinically significant. The alloantibodies were anti-E, anti c, anti e and anti-D. The rate of incidence of these alloantibodies was 3.8%, 1.9%, 0.9% and 0.9% respectively. There was a significant association between alloantibody formation with number of transfused packed red cells (Mann-Whitney Test: p value = 0.035) and age at first transfusion (p value = 0.001). The factors having no association with alloimmunization to red cell antigens are age and gender.Conclusions: Alloimmunization to various erythrocyte blood group antigens is a common problem in multi-transfused β-thalassaemia patients. There is an association between number of transfused packed red cells and age at first transfusion with alloantibody formation in the study.

Assessment of bone mineral density in multi-transfused thalassemia patients in a tertiary care hospital

Objective: Patients with transfusion-dependent thalassemia (TDT) are susceptible to osteopenia and osteoporosis, the mechanism is multi-factorial. Transfusion-related iron overload in endocrine organs leads to impaired hormonal secretion, all contributing to bone loss. The aim is the assessment of bone loss frequency and related parameters affecting it in TDT patients in tertiary care Centre, Jamnagar. Design: Observational cross-sectional study. Subjects: 33. Methods: The study is conducted over 1 year in TDT patients attending G.G.G. hospital for blood transfusion. After clearance from the ethical committee and consent from parents, patients selected according to inclusion criteria and scanned for Bone Mineral Density (BMD) and biochemical tests. Results analyzed with the help of standard statistical tests. Results: Out of 33 patients, 36% (12) patients had osteopenia while 45% (15) patients had osteoporosis. 84% (16) of 19 male patients and 78% (11) of 14 female patients had low BMD. 80% (4) of 5 patients with high S.TSH levels and all patients with low calcium levels, were detected with low BMD.11% (3) of 27 patients having low BMD, had high S. Alkaline phosphatase level. 85% (6) of 7 patients with low S.FSH levels and 73% (8) of 11 patients with low S.L.H. levels have low BMD. 87% (14) of 16 patients with short stature have low bone mass. Conclusion: Evaluating BMD annually in TDT patients helps prevent and intervene timely. Administration of bisphosphonates, calcium, vitamin-D supplements, hormone replacement therapy, bone-forming agents and newer therapies will help improve bone mass. Changing lifestyle with appropriate diet and exercise, iron chelation therapy, early diagnosis and treatment of endocrine insufficiency and regular blood transfusions help achieve optimal bone density.