Guidelines for screening, diagnosis and management of hemoglobinopathies.

The b‑thalassemias and sickle cell disorders are a major health burden in India. Diagnosis and management of these disorders both in adults and in newborns using appropriate approaches and uniform technology are important in different regions of a vast and diverse country as India. In view of a National Thalassemia Control Program to be launched soon, a need was felt for guidelines on whom to screen, cost‑effective technologies that are to be used as well as for establishing prenatal diagnosis programs in regional centers. Newborn screening for sickle cell disorders is in its infancy in India and uniform approaches need to be followed. Also, included are guidelines for monitoring and managing patients who are now growing older and need comprehensive care as well as management of complications of the disease.

Serial assessment of biochemical parameters of red cell preparations to evaluate safety for neonatal transfusions.

Background & Objectives: Neonatologists often prefer fresh blood (<7 days) for neonatal transfusions. The main concerns for stored RBCs are ex vivo storage lesions that undermine red cell functions and may affect metabolic status of neonatal recipients. This study was designed to evaluate serial in vitro changes of biochemical parameters in different RBC preparations during storage to consider for neonatal transfusions even after storage beyond one week. Methods: Twenty five units each of whole blood (CPDA-1 RBC, SAGM RBC) were selected for serial biochemical parameter assessment after each fulfilled the quality criteria (volume and haematocrit). These units were tested serially for supernatant potassium, pH, lactate, haemoglobin, glucose and red cell 2,3 diphosphoglycerate (2,3 DPG) up to 21 days of storage. Results: Within each group of RBC, rise in mean concentration of potassium, lactate and plasma haemoglobin from day 1 to 21 of storage was significant in CPDA-1 RBC having the highest levels at day 21. From day 3 to 21, SAGM RBC had higher mean pH value than CPDA-1 RBC though this difference was not statistically significant. SAGM RBC had highest mean glucose concentration during storage than other two types of red cell preparations (P<0.005). Within each group, fall in mean 2,3 DPG concentration from day 1 to 7 was significant (P<0.05). A positive correlation existed between mean plasma potassium and haemoglobin in all three types of red cells (r=0.726, 0.419, 0.605 for CPDA-1 RBC, SAGM RBC and whole blood respectively, P<0.005). Interpretation & Conclusions: All the three red cell preparations tested revealed biochemical changes within acceptable limits of safety till 21 days of storage. CPDA-1 RBCs had the highest degree of these changes.

Transfusion related acute lung injury.

Transfusion related acute lung injury (TRALI) is an uncommon but potentially fatal adverse reaction to transfusion of plasma containing blood components. We describe a case of 10-year-old male child with aplastic anemia, platelet count of 7800/΅l, B positive blood group who developed fever (39.2΀C), difficulty in breathing and cyanosis within 2 hrs after transfusion of a random platelet concentrate. Despite the best resuscitative efforts, the child died within next 24 hrs. The present case highlights the fact that TRALI should be kept as a differential diagnosis in all patients developing acute respiratory discomfort within 6 hrs of transfusion. Without a 'gold standard' the diagnosis of TRALI relies on a high index of suspicion and on excluding other types of transfusion reactions. Notification to transfusion services is crucial to ensure that a proper investigation is carried out and at-risk donor and recipients can be identified, and risk reduction measures can be adopted.

Evaluation of iron stores in blood donors by serum ferritin.

BACKGROUND & OBJECTIVES: Regular blood donation can lead to pre-clinical iron deficiency as well as iron deficiency anaemia. There is a need to increase the national voluntary blood donation for safe blood supply. However, there is paucity of data in the country regarding impact of regular voluntary blood donation on iron status of donors. Hence, iron stores were evaluated by serum ferritin estimation in the voluntary blood donors at Chandigarh. METHODS: 400 voluntary blood donors included in the study were divided into four groups depending upon their periodicity of blood donations. Pre-donation haemoglobin assessment was done by copper sulphate method. Serum ferritin was estimated by indirect ELISA. RESULTS: The number of female donors with deficient iron stores was more as compared to male donors. First time donors had higher mean serum ferritin levels than that in repeat donors. The frequency of donations per year was more predictive of decreased iron stores rather than the number of lifetime donations. An increase in donation frequency was accompanied by a significant decrease in serum ferritin; values <15 microg/l were found in 21 and 46 per cent of male and female donors respectively who donated once per year, in 29 and 27 per cent in those who donated twice per year and in 49 and 100 per cent in those who donated thrice per year. INTERPRETATION & CONCLUSION: Haemoglobin estimation alone in regular blood donors may not be adequate; serum ferritin estimations may need to be done to detect pre-clinical iron deficiency states. Also, iron supplementation needs to be considered in regular, repeat voluntary blood donors.

Feasibility of pre-operative autologous blood donation in Indian patients with elective orthopaedic surgery.

BACKGROUND & OBJECTIVES: Pre-operative autologous blood donation (PABD) in elective orthopaedic surgeries is a well known procedure in the West. We initiated this programme at a tertiary care hospital in north India to study its feasibility in Indian patients. METHODS: In a prospective case-control study, 144 patients undergoing primary total hip or knee replacement, inter-vertebral discectomy, mal-union and non-union reconstruction were educated and motivated to pre-donate. Patients fulfilling the inclusion criteria and making autologous donation formed the PABD group (n=22). Patients eligible for PABD, but unwilling to participate; age, sex, pre-operative haemoglobin and operative procedure matched acted as controls (n=27). Unit(s) collected was processed like an allogeneic unit. Unit(s) found reactive for infectious markers or not utilized was discarded. Mean blood losses, transfusion trigger, allogeneic exposure and wastage between the two groups were compared. RESULTS: Of the 144 patients motivated, 40 per cent of the eligible subjects pre-deposited. The main motivational factor was fear of getting infection from someone's blood. Cardiac events and anaemia prevented 61.8 per cent patients to participate. Of the 50 units ordered, autologous units with a mean of 1.4 units/patient contributed 62 per cent. For total hip and total knee replacement (THR and TKR), autologous units met 76.2 and 80 per cent respectively of the total blood requirement. A significant decrease in the allogeneic exposure was observed between PABD and control group (18.2 vs 66.7%); 32.3 per cent of the autologous units were discarded. INTERPRETATION & CONCLUSION: Comprehensive PABD programme may be an effective method for reducing the need for allogeneic transfusion in patients undergoing joint replacement surgeries in our country, where transfusion transmitted infections due to high percentage of replacement donations and lack of sensitive assays for testing are still a cause for concern.

Prevalence & significance of hepatitis C virus (HCV) seropositivity in blood donors.

BACKGROUND & OBJECTIVES: The clinical significance of anti HCV antibodies in healthy blood donors remains uncertain. These donors are usually asymptomatic and it is difficult to elicit risk factors of acquiring HCV infection during pre-donation questioning. Limited information on donor recall and follow up studies on anti HCV positive blood donors have been reported from India. Paucity of data which is likely to have an impact on safe blood transfusion programme has prompted us to undertake this study to assess the significance of HCV seropositivity in blood donors with respect to their clinical, biochemical and virological profile. METHODS: A total of 16,250 blood units were screened for the mandatory tests using third generation ELISA (anti HIV 1&2, anti HCV, HBsAg), VDRL and peripheral smear for malaria. Donors reactive for anti HCV were informed. Repeat anti HCV reactive donors were subjected to detailed clinical history focusing on risk factors for HCV transmission. The blood tests included liver function tests (LFT), coagulation and autoimmune profile, qualitative serum cryoglobulins and HCV RNA detection. These donors were followed at 2-3 monthly intervals for a minimum period of six months by LFT. RESULTS: An overall seropositivity of 0.44 per cent (72/16,250) was observed in our donors which was significantly lower in first time, young voluntary donors as compared to replacement donors (0.27 vs. 0.60%). In contrast to drug abuse (6.4%) we found minor percutaneous routes like sharing of shaving kits or visit to a road side barber (32%) as the major risk factor for HCV transmission. There was no prior history of blood transfusion in any of these donors; however history of some surgical procedures was present in 25.8 per cent. Raised transaminases and HCV viraemia were observed in 87 and 71 per cent donors respectively. An association was observed between HCV RNA when the ELISA ratio was >5. INTERPRETATION & CONCLUSION: Voluntary donors form a safe source of blood supply and efforts should be made to increase this precious source to 100 per cent. Abbreviated behavioural donor screening questionnaire for repeat donors is not advisable. Awareness and education of donors is required regarding modes of HCV transmission. HCV positive donors should be informed about their disease, counselled and referred to hepatologist, and permanently deferred for future donations.

Anti Kell allo-antibody in a thalassaemic.

We describe a case of incidental detection of anti Kell antibody in a child with transfusion dependent thalassaemia. Kell antibody detection may be missed by routine indirect antiglobulin test (IAT) crossmatch procedure because of low prevalence of Kell antigen in the general population. A false negative result can be avoided by using sensitive cross matching techniques and screening cells representing antigens in homozygous state, against all clinically significant antibodies. A transfusion alert card describing the nature of antibody and future transfusion policy should be given to such allo-immunized patients.

Platelet alloimmunization in multitransfused patients with haemato-oncological disorders.

BACKGROUND: We studied the incidence of platelet alloimmunization in multitransfused patients with haemato-oncological disorders and determined the factors influencing alloimmunization. We also assessed the effect of alloimmunization on response to platelet transfusion. METHODS: Fifty patients with haemato-oncological disorders who received multiple transfusions were included. The patients were tested for antibodies before they received any transfusion and then after 3-4 weeks of transfusion. Lymphocytotoxicity and platelet immunofluorescence suspension tests were used to detect antiplatelet antibodies. Symptomatic improvement was used to assess the response to platelet transfusions. RESULTS: Thirty patients were positive by the lymphocytotoxicity test, giving an incidence of 60% for anti-HLA antibodies. The panel reactivity of the antibodies ranged from 3% to 100%. Nineteen patients were positive by the platelet immunofluorescence suspension test, 16 of whom were also positive by the lymphocytotoxicity test. The overall incidence of antiplatelet antibodies was 66%. The number of transfusions received and the underlying haemato-oncological disorder were not risk factors for the development of antibodies. Patients with a past history of transfusions and those with a positive obstetric history had a significantly higher incidence of antibodies. The response to transfusion therapy was poor in patients with antibodies, as 71.4% of patients with antibodies were nonresponsive compared to only 26.6% of antibody-negative patients. CONCLUSION: A high percentage of multitransfused patients developed antiplatelet antibodies. Previous sensitization was an important risk factor for the development of antibodies. Patients with high panel reactivity (HLA) showed non-responsiveness to platelet transfusions. Testing for the presence of antiplatelet antibodies and provision of compatible platelets should be important components in the management of patients with platelet transfusion refractoriness.

Prevalence of markers of transfusion transmissible diseases in voluntary and replacement blood donors.

BACKGROUND: Transfusion of safe blood requires a safe donor. The voluntary donor movement encompasses the concept of a donor who is free from transfusion transmissible infections. It is now mandatory to screen blood for hepatitis B surface antigen, antibodies to HIV-1 and HIV-2, antibodies to hepatitis C virus, syphilis and malarial parasites. METHODS: Between 1996 and 2002, 235 461 donors were screened for markers of hepatitis B virus, and HIV-1 and HIV-2 using commercially available ELISA kits, VDRL test for syphilis and Geimsa stain for the malarial parasite, respectively. A total of 56 476 donors were screened for hepatitis C virus antibodies from June 2001 to December 2002, using third-generation ELISA kits. RESULTS: The proportion of voluntary donors increased from 47% to 56% during the study period. The prevalence of HIV showed a steady increase from 0.16% in 1996 to 0.3% in 2002. The prevalence of hepatitis B surface antigen decreased from 1.55% to 0.99%. VDRL reactivity did not show any trend and ranged between 0.11% and 0.66%. Hepatitis C virus antibodies showed a prevalence of 0.4%. The prevalence of all markers was significantly less in voluntary donors. Among the voluntary donors, transfusion transmissible disease markers were significantly less in student donors as compared to other donors. CONCLUSION: A change-over to a voluntary donor service would considerably reduce the number of infectious donors and, among voluntary donors, student donors are the safest.

Congenital dyserythropoietic anemia: clinical and hematological profile.

Congenital dyserythropoietic anemia (CDA) is a rare disorder, which manifests clinically with varying degrees of anemia and hepatosplenomegaly. These features are not pathognomic and a diagnosis of CDA is generally considered after other causes of chronic hemolytic anemia have been ruled out. The clinico-hematological profile of 10 patients with CDA is presented in this communication. Six patients had CDA type II and four had CDA type I. Age at onset of pallor ranged from birth to 9 years. Blood transfusion requirements varied from nil to monthly. This is the first report of CDA type I from India.

Bone marrow trephine biopsy as an adjunct to bone marrow aspiration.

OBJECTIVE: To determine the relative efficacy of bone marrow aspiration as compared to that of trephine biopsy. METHODS: Bone marrow aspiration and bilateral trephine biopsies were performed in 420 consecutive cases. The diagnosis and findings made on bone marrow aspiration were compared with that made on trephine biopsy in each case. RESULTS: Aspiration alone was sufficient in making a diagnosis in 372 (88.6%) cases as it correlated well with the diagnosis made on trephine sections. In the remaining 48 (11.4%) cases trephine biopsy was necessary for making a diagnosis due to incomplete information provided by aspiration or its inability to give a correct diagnosis. These cases were mostly hypoplastic/aplastic marrow, myelofibrosis and marrow involvement by metastatic tumour and lymphomatous infiltration. Often a bilateral marrow biopsy picked up the diagnostic lesion. CONCLUSION: The decision to perform a marrow aspiration alone or in combination with marrow biopsy depends on the diagnosis being considered. In nutritional anaemias, most hematologic malignancies and immune thrombocytopenias, marrow aspiration alone is sufficient, but for detection of disorders with focal marrow involvement bilateral marrow biopsies are a must.