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1.  Response of cardiac endothelial nitric oxide synthase to plasma viscosity modulation in acute isovolemic hemodilution 
Background:
Endothelial nitric oxide synthase (eNOS) is generally expressed in endocardial cells, vascular endothelial cells and ventricular myocytes. However, there is no experimental study elucidating the relationship between cardiac eNOS expression and elevated plasma viscosity in low oxygen delivery pathological conditions such as hemorrhagic shock-resuscitation and hemodilution. This study tested the hypothesis that elevated plasma viscosity increases cardiac eNOS expression in a hemodilution model, leading to positive effects on cardiac performance.
Materials and Methods:
Two groups of golden Syrian hamster underwent an acute isovolemic hemodilution where 40% of blood volume was exchanged with 2% (low-viscogenic plasma expander [LVPE]) or 6% (high-viscogenic plasma expander [HVPE]) of dextran 2000 kDa. In control group, experiment was performed without hemodilution. All groups were performed in awake condition. Experimental parameters, i.e., mean arterial blood pressure (MAP), heart rate, hematocrit, blood gas content and viscosity, were measured. The eNOS expression was evaluated by eNOS Western blot analysis.
Results:
After hemodilution, MAP decreased to 72% and 93% of baseline in the LVPE and HVPE, respectively. Furthermore, pO2 in the LVPE group increased highest among the groups. Plasma viscosity in the HVPE group was significantly higher than that in control and LVPE groups. The expression of eNOS in the HVPE group showed higher intensity compared to other groups, especially compared with the control group.
Conclusion:
Our results demonstrated that cardiac eNOS has responded to plasma viscosity modulation with HVPE and LVPE. This particularly supports the previous studies that revealed the positive effects on cardiac function in animals hemodiluted with HVPE.
doi:10.4103/0973-6247.137444
PMCID: PMC4140050  PMID: 25161348
Cardiac endothelial nitric oxide synthase; hemodilution; plasma expander; plasma viscosity
2.  Incompatible blood transfusion: Challenging yet lifesaving in the management of acute severe autoimmune hemolytic anemia 
Background and Aim:
Autoimmune hemolytic anemia (AIHA) is characterized by the production of autoantibodies directed against red cell antigens. Most patients of AIHA arrive in the emergency or out-patient department (OPD) with severe anemia requiring urgent blood transfusion. Here we share our experience of managing these patients with incompatible blood transfusions and suggest the minimal test required to assure patient safety.
Materials and Methods:
A total of 14 patients admitted with severe anemia, diagnosed with AIHA and requiring blood transfusion urgently were included in the study. A series of immunohematological investigations were performed to confirm the diagnosis and issue best match packed red blood cells (PRBC) to these patients.
Results:
A total of 167 PRBC units were crossmatched for 14 patients of which 46 units (28%) were found to be best match ones and 26 (56.5%) of these units were transfused. A mean turn around time of 222 min was observed in issuing the “best match” blood. Severe hemolysis was observed in all patients with a median hemoglobin increment of 0.88 g/dl after each unit PRBC transfusion.
Conclusion:
Decision to transfuse in AIHA should be based on the clinical condition of the patient. No critical patient should be denied blood transfusion due to serological incompatibility. Minimum investigations such as direct antiglobulin test (DAT), antibody screening and autocontrol should be performed to ensure transfusion safety in patients. All transfusion services should be capable of issuing “best match” PRBCs in AIHA.
doi:10.4103/0973-6247.137445
PMCID: PMC4140051  PMID: 25161349
Alloantibody; autoantibody; autoimmune hemolytic anemia; best match blood; gel technology
3.  Treponema pallidum hemagglutination assay seroreactivity among healthy Indian donors and its association with other transfusion transmitted diseases 
Background:
The aim of the present study was to determine the prevalence of syphilis infection by Treponema pallidum hemagglutination assay (TPHA) among blood donors in Delhi and to study their correlation with other markers of transfusion transmitted infections such as hepatitis C virus (HCV), human immunodeficiency virus (HIV) and hepatitis B surface antigen (HBsAg) so as to establish the utility of TPHA over and above venereal diseases research laboratory test (VDRL), not only as a marker for testing T. pallidum infection, but also as a marker of high risk behavior.
Materials and Methods:
This prospective study was carried out in the Regional Blood Transfusion Centre, Lady Hardinge Medical College and associated Sucheta Kriplani Hospital, New Delhi for a period of 2 years. Donated blood was screened for TPHA seroreactivity along with screening for anti HIV I and II, anti-HCV, HBsAg by third generation enzyme-linked immunosorbent assay test. A total of 8082 serum samples of blood donors were collected from healthy blood donors in our blood bank. They were classified into two groups- test group and control group based on TPHA positivity. The co-occurrence of HBsAg, HIV and HCV infection were determined in TPHA positive blood donors (test group) in comparison with TPHA negative blood donors (control group).
Results:
We found the TPHA seroreactivity to be 4.4% in Delhi's blood donors. Nearly 8.2% (663/8082) of the donated blood had serological evidence of infection by at least one pathogen (syphilis/HIV/hepatitis B virus/HCV) and 6.63% (44/663) donors with positive serology had multiple infections (two or more). Quadruple infection was seen in one donor, triple infection was seen in three donors and double infection was seen in 40 donors. Prevalence of HIV seroreactivity was found to be statistically significant and HCV seroreactivity statistically insignificant in TPHA positive group in comparison to TPHA negative group.
Discussion:
In our study, the TPHA seropositivity correlated with higher HIV and HCV seropositivity and the same correlation has been observed by several other studies also. In view of these observations, we propose that testing for syphilis by more sensitive and specific treponemal markers like TPHA rather than VDRL, rapid plasma reagin tests; as TPHA also has the added advantage of picking up all the high risk donors, whereas, VDRL picks up only currently infected donors. Moreover, TPHA should be continued as a marker of high risk behavior especially in high prevalence areas like India where we don’t have universal access to markers like nucleic acid amplification technique.
doi:10.4103/0973-6247.137447
PMCID: PMC4140052  PMID: 25161350
Syphilis; Treponema pallidum hemagglutination assay; transfusion transmitted infections; Co-infection
4.  Hemolytic disease of fetus and newborn due to maternal red blood cell alloantibodies in the Malay population 
Background:
Maternal red blood cell (RBC) alloimmunization may lead to production of harmful antibodies that result in hemolytic disease of fetus and newborn (HDFN). There is insufficient data on the prevalence of HDFN due to RBC alloantibodies in the Malay neonatal population.
Aim:
The aim of this study was to determine the incidence of HDFN in the Malay neonatal population due to clinically significant RBC alloantibodies.
Subjects and Methods:
A cross sectional study was conducted in Transfusion Medicine Unit, Hospital Universitiy Sains Malaysia over one year period from January to December 2009. A total of 5163 Malay pregnant women who attended labor room for delivery were collected and analyzed prospectively. The blood samples were subjected to the standard immunohematological procedure for RBC antibody screening and identification using reagents of Diamed-ID Gel microtyping system. All the newborns with RBC alloantibody were investigated for the evidence of HDFN.
Results:
Thirty (0.58%) women were found to have clinically significant RBC alloantibodies. Most of the alloantibodies belonged to Rhesus (Rh) system (56.7%) where anti-E (33.3%) was the most common followed by anti-D (10.0%). Rh antibodies were the main cause of HDFN in fourteen (0.27%) neonates. Anti-D and anti-c were identified to cause moderate to very severe HDFN.
Conclusions:
With the low prevalence of clinically significant RBC alloantibodies and HDFN, routine antenatal antibody screening practice may not be advised as a routine practice at present, preferably reserved for those women of RhD negative or with history of HDFN, significantly of those attributed to anti-c.
doi:10.4103/0973-6247.137449
PMCID: PMC4140053  PMID: 25161351
Clinically significant alloantibodies; HDFN; Malay
5.  A survey of resident physicians’ knowledge concerning transfusion medicine in Shiraz, Iran 
Objective:
The knowledge of physicians concerning blood transfusion has a significant impact on the optimal use of blood and blood products. The aim was to survey their knowledge regarding this area and identify whether additional training is required.
Material and Methods:
This cross-sectional study was conducted on all 1st year resident physicians at Shiraz University of Medical Sciences, Iran in 2011. The questionnaire solicited information on demographic variables, knowledge regarding transfusion medicine, education and experience regarding blood transfusion.
Results:
The mean total knowledge score regarding transfusion medicine was 15.44 ± 3.3 (7-25) out of 29. Only about one-fourth (27.4%) replied correctly to over 60% of questions. The mean score of knowledge was higher among residents who stated that they received special training regarding blood transfusion in their medical courses (P < 0.01). Seventy-five percent of residents believed that they had received insufficient education and 97.8% believed that they need additional training.
Conclusion:
The results reflect the uncertainties among resident physicians regarding blood transfusion. It has been suggested that a special transfusion medicine educational program should be added to the medical education curriculum.
doi:10.4103/0973-6247.137451
PMCID: PMC4140054  PMID: 25161352
Blood transfusion; education; knowledge; medical education; physician; transfusion medicine
6.  ABO and Rh (D) group distribution and gene frequency; the first multicentric study in India 
Background and Objectives:
The study was undertaken with the objective to provide data on the ABO and Rh(D) blood group distribution and gene frequency across India.
Materials and Methods:
A total of 10,000 healthy blood donors donating in blood banks situated in five different geographical regions of the country (North, South, East and Center) were included in the study. ABO and Rh (D) grouping was performed on all these samples. Data on the frequency of ABO and Rh(D) blood groups was reported in simple numbers and percentages.
Results:
The study showed that O was the most common blood group (37.12%) in the country closely followed by B at 32.26%, followed by A at 22.88% while AB was the least prevalent group at 7.74%. 94.61% of the donor population was Rh positive and the rest were Rh negative. Regional variations were observed in the distribution. Using the maximum likelihood method, the frequencies of the IA, IB and IO alleles were calculated and tested according to the Hardy Weinberg law of Equilibrium. The calculated gene frequencies are 0.1653 for IA (p), 0.2254 for IB (q) and 0.6093 for IO (r). In Indian Population, O (r) records the highest value followed by B (q) and A (p); O > B > A.
Conclusion:
The study provides information about the relative distribution of various alleles in the Indian population both on a pan-India basis as well as region-wise. This vital information may be helpful in planning for future health challenges, particularly planning with regards to blood transfusion services.
doi:10.4103/0973-6247.137452
PMCID: PMC4140055  PMID: 25161353
ABO; blood group; rhesus
7.  Prevalence of platelet reactive antibodies in patient's refractory to platelet transfusions 
Introduction & Aims:
Though platelet transfusions have greatly reduced the incidence of major haemorrhagic complications associated with the management of haematological and oncological disorders, refractoriness to infused platelets becomes a major clinical problem for many of these patients.
Materials and methods:
The present study was done to determine the percentage of platelet alloimmunisation due to platelet-reactive antibodies in 340 patients with hematologic or oncologic diseases who had received multiple transfusions (> 10) of blood and blood components and showed platelet refractoriness in 1-hour post transfusion sample.
Results:
Platelet-reactive antibodies were detected in the sera of 127 out of 340 patients (37.35%) who received multiple transfusions (> 10) and showed platelet refractoriness.
Conclusion:
Platelet-reactive antibodies appear to be an important cause of platelet refractoriness in patients of acute leukaemia, aplastic anaemia, NHL, MDS and multiple myeloma receiving multiple platelet transfusions. Platelet refractoriness in patients of ITP and chronic leukaemia appears to be due to other causes and not due to platelet-reactive antibodies.
doi:10.4103/0973-6247.137453
PMCID: PMC4140056  PMID: 25161354
Platelet alloimmunization; platelet immunofluorescent test; refractoriness
8.  Response to post-donation counseling is still a challenge in outdoor voluntary blood donation camps: A survey from a tertiary care regional blood center in Eastern India 
Background:
Blood transfusion carries the risk of transmission of several infectious agents. The latest method for blood screening, nucleic acid testing is not affordable in developing countries.
Aim:
The study was aimed to find response to post-donation counseling for reactive markers among the voluntary blood donors donating in blood donation camps.
Material and Methods:
This 1 year study was conducted in 2011. Transfusion transmitted infections testing was performed by routine enzyme linked immunosorbent assay method. The initial human immunodeficiency virus (HIV) reactive donors who returned back to the blood bank were confidentially counseled and referred to the Integrated Counseling Cum Testing Center (ICTC). The hepatitis B surface antigen (HBsAg) and anti-hepatitis C virus (HCV) reactive donors were referred to the gastroenterology department for confirmation by qualitative polymerase chain reaction (PCR, Roche Diagnostics, Germany) and followed-up.
Results:
Twenty seven thousand two hundred forty six 27,246 units were collected during the survey. One hundred twenty nine129 units were reactive for HIV 1 and 2, 99 were reactive for HCV, 206 for hepatitis B virus (HBV). Of these reactive donors, 138 could be personally communicated. Out of 47, 27 donors who returned for counseling were initially reactive for HIV 1 and 2, 8 for HBsAg and 12 for anti-HCV. Two were positive for HBV deoxyribonucleic acid and one was positive for HCV ribonucleic acid. The HIV positivity was detected in 1 of 27 donors at ICTC.
Conclusion:
The response to the post-donation counseling appears in this study to be only 34% (47/138), which is still a challenge.
doi:10.4103/0973-6247.137437
PMCID: PMC4140068  PMID: 25161343
Integrated counseling and testing center; post-donation counseling; transfusion transmitted infections
9.  Alloimmunization and autoimmunization in transfusion dependent thalassemia major patients: Study on 319 patients 
Background:
The development of anti-red blood cell antibodies (both allo-and autoantibodies) remains a major problem in thalassemia major patients. We studied the frequency of red blood cell (RBC) alloimmunization and autoimmunization among thalassemia patients who received regular transfusions at our center and analyzed the factors, which may be responsible for development of these antibodies.
Materials and Methods:
The study was carried out on 319 multiply transfused patients with β-thalassemia major registered with thalassemia clinic at our institute. Clinical and transfusion records of all the patients were examined for age of patients, age at initiation of transfusion therapy, total number of blood units transfused, transfusion interval, status of splenectomy or other interventions. Alloantibody screening and identification was done using three cell and 11 cell panel (Diapanel, Bio-rad, Switzerland) respectively. To detect autoantibodies, autocontrol was carried out using polyspecific coombs (IgG + C3d) gel cards.
Results:
Eighteen patients out of total 319 patients (5.64%) developed alloantibodies and 90 (28.2%) developed autoantibodies. Nine out of 18 patients with alloantibodies also had autoantibodies. Age at first transfusion was significantly higher in alloimmunized than non-immunized patients (P = 0.042). Out of 23 alloantibodies, 52.17% belonged to Rh blood group system (Anti-E = 17%, Anti D = 13%, Anti-C = 13%, Anti-Cw = 9%), 35% belonged to Kell blood group system, 9% of Kidd and 4% of Xg blood group system.
Conclusion:
Alloimmunization was detected in 5.64% of multitransfused thalassemia patients. Rh and Kell blood group system antibodies accounted for more than 80% of alloantibodies. This study re-emphasizes the need for RBC antigen typing before first transfusion and issue of antigen matched blood (at least for Rh and Kell antigen). Early institution of transfusion therapy after diagnosis is another means of decreasing alloimmunization.
doi:10.4103/0973-6247.137438
PMCID: PMC4140069  PMID: 25161344
Alloimmunization; autoimmunization; thalassemia major; transfusion
10.  Experience of buffy coat pooling of platelets as a supportive care in thrombocytopenic dengue patients: A prospective study 
Random donor platelet (RDP) is not sufficient to improve the platelet count in most thrombocytopenic patients. Single donor platelet (SDP) or buffy coat pooled platelet (BCPP) are the two choices to provide a full therapeutic dose of platelets. However, there are constraints in the preparation of SDP due to stringent donor selection procedure, time required for procedure, and need of special expensive equipments and kits. BCPP is widely practiced, especially in the European countries, since 1995. In India, we decided to adopt the procedure of buffy coat pooling of platelets, especially for economically backward patients and for emergencies. This study was prospectively conducted from September 2009 to September 2010. A total of 129 units of BCPP [tested prior for viral markers by enzyme-linked immunosorbent assay (ELISA) and individual donor nucleic acid amplification test (ID-NAT)] were issued to 129 patients suffering from dengue and were included in this study. For comparison between efficacy of SDP and BCCP, patients were divided into two groups of 50 each. The post-transfusion platelet counts of the patients were noted after 2 hours of transfusion for each type of component. The platelet yield varied from 2.5 to 4.4 Χ 1011 in BCPP samples. The samples analyzed were sterile without any contamination. The different biochemical parameters were analyzed in detail. The observed post-transfusion platelet recovery and corrected count increment (CCI) at 1 hour and 24 hours after BCPP transfusion were similar to that after SDP transfusion. Hence, we concluded that BCPP can be a low cost alternative to SDP in the times of emergencies like dengue and non-affordability by the patient for SDP.
doi:10.4103/0973-6247.137439
PMCID: PMC4140070  PMID: 25161345
buffy coat; buffy coat pooling of platelets; random donor platelets; single donor platelets
11.  Prospective evaluation of 2% (w/v) alcoholic chlorhexidine gluconate as an antiseptic agent for blood donor arm preparation 
Aim:
A prospective study was undertaken to evaluate the use of 2% (w/v) alcoholic chlorhexidine gluconate (2% AlcCHG) in donor arm preparation, to monitor the contamination rate of blood products after the collection and to find incidence of transfusion associated bacteremia.
Settings and Design:
Optimal skin antisepsis of the phlebotomy site is essential to minimize the risk of contamination. Food and Drug Administration (FDA) in India has recommended antisepsis with three-step regimen of spirit-10% povidone iodine-spirit for donor arm antisepsis, but not with chlorhexidine, which is recommended by many other authors.
Material and Methods:
A total of 795 donors were studied from July 2011 to January 2012. Spirit-10% povidone iodine-spirit was used for 398 donors and 2% AlcCHG was used for 397 donors with the two-step method for arm antisepsis. Swabs were collected before and after use of antiseptic agents for all the donors. All the blood products collected from donors with growth in post-antisepsis swabs were cultured. A total of 123 various blood products were cultured irrespective of the method and result of antisepsis was observed. A total of seven patients had mild transfusion reaction. The transfused blood products, blood and urine specimen of the patients who had transfusion reaction were also cultured.
Results:
Seven donors out of 398 donors had growth in post-antisepsis swab with spirit-10% povidone iodine-spirit protocol and three donors out of 397 donors had growth in post-antisepsis swab with 2% AlcCHG protocol. All blood products collected from donors who had growth in post-antisepsis swabs when cultured had no growth. There was no contamination of blood products.
Conclusions:
Two percent (w/v) alcoholic chlorhexidine gluconate with two-step protocol can be used as an antiseptic agent for donor arm preparation without considerable cost difference. It is at par with spirit 10% povidone iodine spirit protocol as suggested by FDA in India. There was no reported transfusion associated bacteremia in the study period.
doi:10.4103/0973-6247.137441
PMCID: PMC4140071  PMID: 25161346
Bacteremia; blood donor arm; chlorhexidine gluconate
12.  Clinical significance of antibody specificities to M, N and Lewis blood group system  
Context:
The clinically significant antibodies are those active at 37°C and/or by the indirect antiglobulin test. Most of the published literature refers to antibodies of Lewis blood group system to be insignificant, whereas antibodies to M and N blood groups are associated with variable clinical significance.
Aims:
The aim of this study is to find the frequency and clinical significance of antibodies to M, N and Lewis blood group systems.
Settings and Design:
The study was carried out retrospectively from January 2009 to December 2012.
Materials and Methods:
Antibody screening was performed by solid phase red cell adherence (SPRCA) technique using four cell screening panel on a fully automated platform GALILEO (Immucor Inc. USA). In case of a positive antibody screen, antibody identification was performed using SPRCA (GALILEO, Immucor Inc. USA).
Results:
A total of 49,077 red cell antibody screens were performed and a total of 427 identifications of red cell antibodies were carried out. A total of 304 specific antibodies were detected: 8.22% of antibodies were of anti-M specificity and 2.96% were of anti-N specificity. Majority (84%) of anti-M and 77.78% of anti-N were of Immunoglobulin G (IgG) class reacting at 37°C. 1.31% of the antibodies were directed against Lewis system antigens of which 0.65% were anti-Lea and 0.65% were anti-Leb. Half of the Lewis system antibodies, i.e., 1 each of anti-Lea and anti-Leb were of IgG class.
Conclusion:
Our study highlights the importance of detecting the thermal amplitude of antibodies with variable clinical significance especially if both IgG and IgM types of antibodies are associated with it so as to establish their clinical significance.
doi:10.4103/0973-6247.137442
PMCID: PMC4140072  PMID: 25161347
Anti-M; anti-N; clinical significance; Lewis
13.  Sensitivity of individual and mini-pool nucleic acid testing assessed by dilution of hepatitis B nucleic acid testing yield samples 
Introduction:
For nucleic acid testing (NAT) of blood donations, either the blood samples can be pooled together in a batch of six or eight prior to testing (mini-pool-NAT [MP-NAT]), or the tests can be run on every individual sample (individual donor-NAT [ID-NAT]). It has been debated in various studies whether pooling of samples results in decreased sensitivity of detection as the volume of individual samples gets lesser in a pool. The objective of this study was to investigate the effect of dilution on the sensitivity of tests.
Materials and Methods:
The study was performesd on nine plasma samples which were hepatitis B reactive exclusively by Procleix Ultrio Plus and not by Procleix Ultrio or serology. These nine exclusive UltrioPlus ID-NAT yield samples were diluted in 1:2, 1:4. 1:6 and 1:8 dilutions using previously tested negative plasma and each dilution of every sample along with archived undiluted sample were retested in three replicates with Procleix Ultrio Plus Assay.
Results:
Among NAT yield samples, 88.88% of the samples were detected when retested in ID-NAT in undiluted form. Samples with higher viral load (sample 5 and 6) were detected by all dilutions. When samples with viral load below 20 IU/mL were tested in dilutions of 1:6 or 1:8, only 9 out of 27 replicates (33.33%) were detected. This means that more than 67% of low viral load samples were missed by MP-NAT of 1:6 or 1:8 dilution out of total NAT yield samples.
Conclusion:
Individual Donor NAT is ideal methodology for NAT as dilution due to pooling may miss samples with low viral load as evident in this study.
doi:10.4103/0973-6247.126684
PMCID: PMC3943140  PMID: 24678169
Hepatitis B surface antigen; hepatitis B virus; nucleic acid testing; pooling of samples
14.  Hepatitis E seroprevalence among blood donors: A pilot study from Western India 
Background:
Hepatitis E virus (HEV) is emerging as a potential threat to blood safety after several cases of transmission by transfusion or transplantation have been described. Currently, blood donors in India are not screened for HEV. The studies conducted on HEV in recent times in India have focused on epidemiology and future perspectives, but there is no published study on blood donors. To address possible issues surrounding blood safety and risk of HEV transmission within the Indian blood supply, HEV seroprevalence study was conducted in blood donors at our center.
Materials and Methods:
A total of 460 male voluntary blood donors were selected for the study and after taking their written consent. Serum anti-HEV IgM was detected by Dia.Pro HEV kit (Diagnostic Bioprobes Srl, Milano, Italy).
Results:
The study population was composed of 460 male voluntary blood donors and their age ranged from 18 to 60 years with a mean age of 30.48 years. Out of 460 donors, 22 (4.78%) donors were tested positive for IgM anti-HEV and the mean value alanine aminotransferase (ALT) was 26.06 IU/L, the highest being 93.5 IU/L. Normal reference value of ALT in our center was 40 IU/L. Out of 22 anti-HEV positive donors, 19 (86.36%) had ALT values above 40 IU/L.
Conclusion:
HEV seroprevalence of 4.78% in our center. Though reports of HEV transmission through blood has been reported from various parts of the world, before making it as a mandatory screening test among blood donors in India, further studies with confirmatory assay of HEV need to be done.
doi:10.4103/0973-6247.126685
PMCID: PMC3943141  PMID: 24678170
Blood donors; hepatitis E virus; seroprevalence
15.  Hepatitis C virus genotypes among multiply transfused hemoglobinopathy patients from Northern Iraq 
Background and Aim:
Owing to the scarcity of data on hepatitis C virus (HCV) genotypes in Iraq and due to their epidemiological as well as therapy implications, this study was initiated aiming at determining these genotypes in Northern Iraq.
Materials and Methods:
A total of 70 HCV antibody positive multi transfused patients with hemoglobinopathies, who had detectable HCV ribonucleic acid, were recruited for genotyping using genotype-specific nested polymerase chain reaction.
Results:
The most frequent genotype detected was genotype 4 (52.9%) followed by 3a (17.1%), 1b (12.9%) and 1a (1.4%), while mixed genotypes (4 with either 3a or 1b) were detected in 7.1%.
Conclusion:
The predominance of genotype 4 is similar to other studies from surrounding Eastern Mediterranean Arab countries and to the only earlier study from central Iraq, however the significant high proportion of 3a and scarcity of 1a, are in contrast to the latter study and may be explainable by the differing population interactions in this part of Iraq. This study complements previous studies from Eastern Mediterranean region and demonstrates relative heterogeneity of HCV genotype distribution within Iraq and should trigger further studies in other parts of the country.
doi:10.4103/0973-6247.126687
PMCID: PMC3943142  PMID: 24678171
Genotype 4; genotyping; hepatitis C virus; Iraq
16.  Half a decade of mini-pool nucleic acid testing: Cost-effective way for improving blood safety in India 
Background and Objectives:
It is well established that Nucleic acid testing (NAT) reduces window phase of transfusion transmissible infections (TTI) and helps improve blood safety. NAT testing can be done individually or in pools. The objectives of this study were to determine the utility, feasibility and cost effectiveness of an in-house minipool-NAT(MP-NAT).
Materials and Methods:
Blood donors were screened by history, tested by ELISA and sero-negative samples were subjected to an in-house NAT by using reverse transcriptase-polymerase chain reaction (RT-PCR). Testing was done in mini-pools of size eight (8). Positive pools were repeated with individual samples.
Results:
During the study period of Oct 2005-Sept 2010 (5 years) all blood donors (n=53729) were screened by ELISA. Of which 469 (0.87%) were positive for HIV-1, HBV or HCV. Sero-negative samples (n=53260) were screened by in-house MP-NAT. HIV-NAT yield was 1/53260 (n=1) and HBV NAT yield (n=2) was 1/26630.
Conclusion:
NAT yield was lower than other India studies possibly due to the lower sero-reactivity amongst our donors. Nevertheless it intercepted 9 lives including the components prepared. The in-house assay met our objective of improving blood safety at nominal cost and showed that it is feasible to set up small molecular biology units in medium-large sized blood banks and deliver blood within 24-48 hours. The utility of NAT (NAT yield) will vary based on the donor population, the type of serological test used, the nature of kit employed and the sensitivity of NAT test used. The limitations of our in-house MP-NAT consisted of stringent sample preparation requirements, with labor and time involved. The benefits of our MP-NAT were that it acted as a second level of check for ELISA tests, was relatively inexpensive compared to ID-NAT and did not need sophisticated equipment.
doi:10.4103/0973-6247.126688
PMCID: PMC3943143  PMID: 24678172
Blood donor testing; blood safety; in house assay; mini-pool nucleic acid testing; real-time-polymerase chain reaction
17.  Knowledge, attitude, and beliefs of young, college student blood donors about Human immunodeficiency virus 
Introduction:
Young people, who tend to be healthy, idealistic, and motivated, are an excellent pool of potential voluntary unpaid blood donors. Recruiting and retaining young blood donors improves the long term safety and sufficiency of a country's blood supply. Knowledge, attitude, and beliefs about Human immunodeficiency virus (HIV) should play an important role in prevention of disease transmission.
Materials and Methods:
This study was a questionnaire based survey, conducted to explore the levels of knowledge, attitude, and beliefs about HIV in young college student blood donors.
Results:
The results showed that the proportion of participants with comprehensive knowledge of HIV prevention and transmission was lesser than expected. Increase in education level and male gender was found to be significantly associated with high HIV-related knowledge. The responses on the different aspects of HIV-related attitude were also varied and there is still stigma associated with Acquired Immunodeficiency Syndrome (AIDS) even in the educated groups.
Discussion:
There was a spectrum of myths and misperceptions emphasizing the need of education that recognizes the social context of attitude towards HIV. Results from this study may contribute to the development of appropriate educational and training material for this group of donors which in turn, may assist in achieving the elusive goal of safe blood supply in future.
doi:10.4103/0973-6247.126689
PMCID: PMC3943144  PMID: 24678173
Attitude; beliefs; blood donors; college students; HIV; knowledge
18.  A single-centre study of vasovagal reaction in blood donors: Influence of age, sex, donation status, weight, total blood volume and volume of blood collected 
Context:
Vasovagal reactions (VVRs) in blood donors.
Aim:
To find an association of age, sex, donation status, weight, total blood volume and volume of blood collected with occurrence of immediate VVR.
Settings and Design:
Retrospective single-centre study.
Materials and Methods:
The study was conducted from March 2000 to November 2010 at a tertiary care blood transfusion centre. All VVRs with or without syncope occurring during or at the end of donation were noted.
Statistical Analysis Used:
For qualitative association, c2-test was used. Unpaired ‘t’ test was used for assessing difference between two groups with respect to VVR status. Simultaneous impact of all risk factors was assessed using multivariate logistic regression analysis. The data entry software SPSS (version 17.0) was used for statistical analysis. A P-value <0.05 was considered statistically significant.
Results:
Overall 1085 VVRs were reported in relation to 88,201 donations, resulting in an overall VVR rate of 1.23%, that is, an incidence of 1 in every 81 donations. Donors with low blood volume, first-time donors, with low weight and female donors had higher absolute donation VVR rates than other donors.
Conclusions:
Donation-related vasovagal syncopal reactions are a multifactorial process determined largely by weight, age, first-time donor status and total blood volume. Our study reinforces the fact that blood donation is a very safe procedure, which could be made even more event-free by following certain friendly, reassuring practices and by ensuring strict pre-donation screening procedures.
doi:10.4103/0973-6247.126690
PMCID: PMC3943146  PMID: 24678174
Blood donor; first-time donors; total blood volume; vasovagal reaction; volume of blood collected
19.  The association of transfusion status with antioxidant enzymes and malondialdehyde level in Nigerians with sickle cell disease 
Background:
Sickle cell disease (SCD) often leads to chronic hemolytic anemia of varying severity, and blood transfusion may be employed in the management of SCD complications.
Objective:
The aim of the study was to evaluate the effect of blood transfusion on the activities of some antioxidant enzymes as well as lipid peroxide and to relate transfusion status to these enzymes and malondialdehyde (MDA) in SCD patients.
Materials and Methods:
Glutathione peroxidase (GPX), superoxide dismutase, catalase, MDA, and lipoproteins were assayed in 87 SCD and 20 age- and sex-matched subjects with normal hemoglobin. Of the 87 SCD patients, 30 had multiple transfusions, 21 had been transfused once while 36 had not been transfused within the last 3 months.
Results:
Statistically significant decrease in the mean levels of GPX (P = 0.045) and Cu/Zn SOD (P = 0.001) and increased (P = 0.001) MDA were observed in the transfused compared to non-transfused patients. Similarly, significant decrease (P = 0.001) in Cu/Zn SOD and increase (P = 0.01) in MDA were observed in multi transfused compared to those who had been transfused once. Transfusion status correlated (P <0.047) inversely with Cu/Zn SOD and positively with MDA.
Conclusion:
Reduced activity levels of serum antioxidant enzymes and increased mean levels of MDA were observed in transfused compared to non-transfused SCD patients and these changes correlated with transfusion status.
doi:10.4103/0973-6247.126692
PMCID: PMC3943147  PMID: 24678175
Antioxidant enzymes; malondialdehyde; sickle cell disease; transfusion
20.  Phenotype frequencies of blood group systems (Rh, Kell, Kidd, Duffy, MNS, P, Lewis, and Lutheran) in blood donors of south Gujarat, India 
Background:
This is the first study on phenotype frequencies of various blood group systems in blood donors of south Gujarat, India using conventional tube technique.
Material and Methods:
A total of 115 “O” blood group donors from three different blood banks of south Gujarat were typed for D, C, c, E, e, K, Jka, Lea, Leb, P1, M, and N antigens using monoclonal antisera and k, Kpa, Kpb, Fya,Fyb, Jkb, S,s, Lua, and Lub antigens were typed using polyclonal antisera employing Indirect Antiglobulin Test. Antigens and phenotype frequencies were expressed as percentages.
Results:
From the 115 blood donor samples used for extended antigen typing in the Rh system, e antigen was found in 100% donors, followed by D [84.35%], C [81.74%], c [56.32%], and E [21.74%] with DCe/DCe (R1 R1, 40.87%) as the most common phenotype. k was found to be positive in 100% of donors and no K+k- phenotype was found in Kell system. For Kidd and Duffy blood group system, Jk(a+b+) and Fy(a-b-) were the most common phenotypes with frequency of 52.17% and 48.69%, respectively. In the MNS system, 39.13% donors were typed as M+N+, 37.39% as M+N-, and 23.48% as M-N+. S+s+ was found in 24.35% of donors, S+s- in 8.69%, and S-s+ as the commonest amongst donors with 66.96%. No Lu(a+b+) or Lu(a+b-) phenotypes were detected in 115 donors typed for Lutheran antigens. A rare Lu(a-b-) phenotype was found in 2.61% donors.
Conclusion:
Data base for antigen frequency of various blood group systems in local donors help provide antigen negative compatible blood units to patients with multiple antibodies in order to formulate in-house red cells for antibody detection and identification and for preparing donor registry for rare blood groups.
doi:10.4103/0973-6247.126693
PMCID: PMC3943149  PMID: 24678176
Blood donors; blood group systems; India; phenotype frequency; red cell antigens; south Gujarat
21.  Retrospective evaluation of adverse transfusion reactions following blood product transfusion from a tertiary care hospital: A preliminary step towards hemovigilance 
Background:
The goal of hemovigilance is to increase the safety and quality of blood transfusion. Identification of the adverse reactions will help in taking appropriate steps to reduce their incidence and make blood transfusion process as safe as possible.
Aims:
To determine the frequency and type of transfusion reactions (TRs) occurring in patients, reported to the blood bank at our institute.
Materials and Methods:
A retrospective review of all TRs reported to the blood bank at the All India Institute of Medical Sciences, between December 2007 and April 2012 was done. All the TRs were evaluated in the blood bank and classified using standard definitions.
Results:
During the study period a total of 380,658 bloods and blood components were issued by our blood bank. Out of the total 196 adverse reactions reported under the hemovigilance system, the most common type of reaction observed was allergic 55.1% (n = 108), followed by febrile non-hemolytic transfusion reaction (FNHTR) 35.7% (n = 70). Other less frequently observed reactions were Anaphylactoid reactions 5.1% (n = 10), Acute non-immune HTRs 2.6% (n = 5), Circulatory overload 0.5% (n = 1), Transfusion related acute lung injury 0.5% (n = 1), Delayed HTRs 0.5% (n = 1). Not a single case of bacterial contamination was observed.
Conclusion:
The frequency of TRs in our patients was found to be 0.05% (196 out of 380,658). This can be an underestimation of the true incidence because of under reporting. It should be the responsibility of the blood transfusion consultant to create awareness amongst their clinical counterpart about safe transfusion practices so that proper hemovigilance system can be achieved to provide better patient care.
doi:10.4103/0973-6247.115564
PMCID: PMC3757769  PMID: 24014939
Adverse transfusion reactions; blood transfusion; hemovigilance
22.  Transfusion transmittable infections – Seroprevalence among blood donors in a tertiary care hospital of Delhi 
Context:
Transfusion transmittable infections (TTI) continue to be a major threat to safe transfusion practices. Blood is one of the major sources of transmission of infectious diseases viz. human immunodeficiency virus (HIV), hepatitis B virus (HBV), hepatitis C virus (HCV), syphilis, malaria, and many other infections in India. Screening assays for the infectious diseases with excellent sensitivity and specificity helps to enhance the safety of the blood transfusions reducing the diagnostic window period as much as possible.
Aims:
The present study was designed to determine the seroprevalence of TTIs viz., HIV, HCV, and HBV, among the blood donors in Max Super Specialty Hospital, New Delhi, India based on dual testing strategy using high sensitive screening assays such as enhanced chemiluminescence assay and nucleic acid testing (NAT).
Materials and Methods:
A total of 41207 blood units collected from the donors (both voluntary and replacement donors) were screened for the TTI s, viz., anti HIV 1 and 2 antibody, anti HCV antibody, anti HBcore antibody, and HBsAg by enhanced chemiluminescence assay on VITROS® ECiQ immunodiagnostics system. NAT was performed using Roche Cobas® TaqScreen MPX assay, which can detect simultaneously HIV 1 (groups M and O), HIV-2, HCV, and HBV on Roche Cobas® s201 system.
Results:
The seroprevalence of HIV, HBsAg, anti HBcore antibody, and HCV based on enhanced chemiluminescence assay was found to be 0.25, 0.2, 7.06, and 0.7%, respectively. A total number of 6587 samples from July 2010 to December 2010 were tested on NAT, of which 3 samples were reactive for HBV in NAT; this was missed by enhanced chemiluminescence assay.
Conclusions:
Based on the seroprevalence study of infectious diseases viz., HIV, HBV, and HCV, we conclude that screening of blood and blood components by dual testing strategy using high sensitivity serological assay like enhanced chemiluminescence technology and NAT helps in detecting the potentially infectious blood units in all phases of infection, which aids in enhancing the safety of blood transfusion and reducing the potential risk of post-transfusion infection.
doi:10.4103/0973-6247.115566
PMCID: PMC3757770  PMID: 24014940
Seroprevalence; seroprevalence in blood donors; transfusion transmittable infections
23.  RHD alleles in the Tunisian population 
Background:
A comprehensive survey of RHD alleles in Tunisia population was lacking. The aim of this study was to use a multiplex RHD typing assay for simultaneous detection of partial D especially with RHD/RHCE deoxyribonucleic acid (DNA) sequence exchange mechanism and some weak D alleles.
Materials and Methods:
Six RHD specific primer sets were designed to amplify RHD exons 3, 4, 5, 6, 7 and 9. DNA from 2000 blood donors (1777 D+ and 223 D-) from several regions was selected for RHD genotyping using a PCR multiplex assay. Further molecular investigations were done to characterize the RHD variants that were identified by the PCR multiplex assay.
Results:
In the 1777 D+ samples, only 10 individuals showed the absence of amplification of exons 4 and 5 that were subsequently identified by PCR-SSP as weak D type 4 variants. No hybrid allele was detected. In the 223 D-, RHD amplification of some exons was observed only in 5 samples: 4 individuals expressed only RHD exon 9, and one subject lacking exons 4 and 5. These samples were then screened by PCR-SSPs on d(C) ces and weak D type 4, respectively.
Conclusion:
The weak D type 4 appears to be the most common D variant allele. We have not found any partial D variant. Findings also indicated that RHD gene deletion is the most prevalent cause of the D- phenotype in the Tunisian population.
doi:10.4103/0973-6247.115568
PMCID: PMC3757771  PMID: 24014941
Partial D; polymerase chain reaction multiplex; polymerase chain reaction with sequence-specific priming; RHD genotyping; Tunisia; weak D
24.  Seroprevalence of human immunodeficiency virus in north Indian blood donors using third and fourth generation Enzyme linked immunosorbent assay 
Background:
The percentage of HIV cases attributable to blood transfusion has decreased significantly in the last decade. The newer 4th generation Enzyme linked immunosorbent assay (ELISA) has been shown to have increased sensitivity compared to 3rd generation ELISA.
Objectives:
To estimate the seroprevalence of HIV among blood donors using 4th generation ELISA assay and to compare it with the 3rd generation ELISA.
Materials and Methods:
This prospective study involved 10,200 blood donors- 6,800 were voluntary donors (3400-students and 3400-non students) and 3400 were replacement donors. All blood units were tested with 3rd as well as 4th generation ELISA. All samples found reactive or in grey zone with either 3rd or 4th generation ELISA were retested by Western blot (WB).
Results:
The seroprevalence of HIV was estimated to be 1.37/1000 donations (0.14%) with 3rd generation ELISA compared to 3.62/1000 donations (0.36%) with 4th generation ELISA (p>0.05). The seroprevalence of HIV among voluntary donors was estimated to be 1.32/1000 donations (0.13%) with 3rd generation ELISA and 3.67/1000 donations (0.36%) with 4th generation ELISA. The prevalence of HIV among replacement donors was 1.47/1000 donations (0.15%) with 3rd generation ELISA and 3.52/1000 donations (0.35%) with 4th generation ELISA.
Conclusion:
4th generation HIV ELISA detects a higher number of seroreactive donors compared to 3rd generation ELISA. However, larger studies are required with confirmatory tests for both 3rd and 4th generation ELISA for making any policy changes.
doi:10.4103/0973-6247.115570
PMCID: PMC3757772  PMID: 24014942
Enzyme linked immunosorbent assay; Human immunodeficiency virus; window period; transfusion associated HIV/AIDS; blood donors
25.  Looking to the future, working on the present. Intention to donate blood and risk behavior: A study on adolescents 
Background:
The aim of this work was to investigate adolescents’ propensity to donate blood, the incidence in this population of risk behaviors that limit and restrict the potential for blood donation, thus reducing the number of possible subjects suitable for donation, and the link between propensity to donate and healthy lifestyles.
Materials and Methods:
Adolescents were asked to complete an anonymous self-report questionnaire during class time. The questionnaire included ad hoc items designed to investigate the intention to donate blood and behaviors traditionally considered to put adolescents’ “health at risk”. Participants were 201 students from northern Italy (range 12–19, M = 17.98, SD = 1.82; 38.9% male, 61.1% female).
Results:
About half of respondents perceived that they were able to donate blood and could overcome any obstacle that might prevent them from doing so. Some of the examined risk behaviors were widespread in the population under consideration. The linear regressions run show that there is no link between the propensity to give blood and a tendency to engage in healthy lifestyles.
Conclusion:
The study shows that adolescents demonstrate an interest in blood donation; however, it shows a real lack of “action” to make it happen: there is, indeed, a serious lack of attention to matters related to healthy lifestyles. In fact, adolescents are not aware of how to act to access the world of donation, with the risk that they present themselves at blood donation centers and are then found to be unfit to donate. The research offers many suggestions and implications.
doi:10.4103/0973-6247.115573
PMCID: PMC3757773  PMID: 24014943
Adolescence; blood donation; policy; recruitment; risk behavior

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