Malaysian Blood Donors Research Articles

Characteristics of ABO Antibodies in Group O Malaysian Blood Donors.

Haemolytic transfusion reactions (HTRs) due to anti-A and anti-B antibodies in Group O blood products are rare but potentially fatal. This study aimed to identify the prevalence of high ABO antibody titre and the immunoglobulin (Ig) classes (IgM only or with IgG) and the prevalence of haemolysin antibodies in Group O blood donors. Plasma from Group O blood donors was tested by using antibody titration at room temperature. Titres ≥ 64 were considered high. The plasma was treated with 0.01 M dithiothreitol (DTT) to determine the presence of IgG antibodies and titre. IgG titres ≥ 64 were considered high. Tests for haemolysis were conducted by mixing the plasma with 3% fresh A1 and B cell suspensions and incubating at 37 °C. The haemolysis was observed macroscopically. Of 311 donors, 238 (76.5%) showed high anti-A and/or anti-B antibody titres. The highest antibody titre obtained was 256. Female and younger donors (< 40 years old) had higher anti-A and anti-B titres. The anti-B titre showed an association with gender (P < 0.001), and was high in female donors (77.8%). Males aged over 50 years old were found to have low mean titre antibodies. Most donors had both IgM and IgG ABO antibodies. The prevalence of haemolysins in our population was 3.5%. Most of our O blood donors had a high ABO antibody titre but a low prevalence of haemolysins. Males aged over 50 years old are the best O donors for preventing HTRs, particularly when mismatch transfusion is required. We recommend a transfusion unit screen for ABO antibody titre in younger female donors (< 40 years old), to prevent the transfusion of high titre O blood products into non-O recipients.

Prevalence of GP. Mur variant phenotype among Malaysian blood donors.

A number of glycophorin variant phenotypes or hybrid glycophorin variants of the MNS blood group system bear multiple immunogenic antigens such as Mia, Mur, and MUT. In the East and Southeast Asian populations, glycoprotein (GP.) Mur is the most common glycophorin variant phenotype expressing those three immunogens. The aim of this study was to detect MNS system glycophorin variant phenotypes (GP. Mur, GP. Hop, GP. Bun, GP. HF, and GP. Hut) among Malaysian blood donors. In this cross-sectional study, 144 blood donors were selected under stratified random sampling. The deoxyribonucleic acid was extracted from whole blood samples, followed by a polymerase chain reaction assay. Sanger sequencing was used to identify the specific MNS variants and then validated by a serological crossmatch with known anti-Mur and anti-MUT. GP. Mur was identified among Malaysian blood donors with a prevalence of 6.94%, and no other variants of the MNS system were found. The present study substantiates that GP. Mur is the main variant of the MNS system glycophorin (B-A-B) hybrid in Malaysian blood donors. GP. Mur-negative red blood cells must therefore be considered in the current transfusion policy in order to prevent alloimmunization and immune-mediated transfusion reactions, particularly in transfusion-dependent patients.

Genotype-Phenotype Correlation of β-Thalassemia in Malaysian Population: Toward Effective Genetic Counseling

Effective prevention of β-thalassemia (β-thal) requires strategies to detect at-risk couples. This is the first study attempting to assess the prevalence of silent β-thal carriers in the Malaysian population. Hematological and clinical parameters were evaluated in healthy blood donors and patients with β-thal trait, Hb E (HBB: c.79G>A)/β-thal and β-thal major (β-TM). β-Globin gene sequencing was carried out for 52 healthy blood donors, 48 patients with Hb E/β-thal, 34 patients with β-TM and 38 patients with β-thal trait. The prevalence of silent β-thal carrier phenotypes found in 25.0% of healthy Malaysian blood donors indicates the need for clinician’s awareness of this type in evaluating β-thal in Malaysia. Patients with β-TM present at a significantly younger age at initial diagnosis and require more blood transfusions compared to those with Hb E/β-thal. The time at which genomic DNA was extracted after blood collection, particularly from patients with β-TM and Hb E/β-thal, was found to be an important determinant of the quality of the results of the β-globin sequencing. Public education and communication campaigns are recommended as apparently healthy individuals have few or no symptoms and normal or borderline hematological parameters. β-Globin gene mutation characterization and screening for silent β-thal carriers in regions prevalent with β-thal are recommended to develop more effective genetic counseling and management of β-thal.

Elucidation of Weak D Phenotype among Malaysian Blood Donors using Molecular Basis

Elucidation of Weak D Phenotype among Malaysian Blood Donors using Molecular Basis

Molecular and serological detection of occult hepatitis B virus among healthy hepatitis B surface antigen-negative blood donors in Malaysia.

Occult hepatitis B infections are becoming a major global threat, but the available data on its prevalence in various parts of the world are often divergent. This study aimed to detect occult hepatitis B virus in hepatitis B surface antigen-negative serum using anti-HBc as a marker of previous infection. A total of 1000 randomly selected hepatitis B surface antigen-negative sera from blood donors were tested for hepatitis B core antibody and hepatitis B surface antibody using an ELISA and nested polymerase chain reaction was done using primers specific to the surface gene (S-gene). Of the 1000 samples 55 (5.5%) were found to be reactive, of which 87.3% (48/55) were positive for hepatitis B surface antibody, indicating immunity as a result of previous infection however, that does not exclude active infection with escaped mutant HBV. Nested PCR results showed the presence of hepatitis B viral DNA in all the 55 samples that were positive for core protein, which is in agreement with the hepatitis B surface antibody result. This study reveals the 5.5% prevalence of occult hepatitis B among Malaysian blood donors as well as the reliability of using hepatitis B core antibody in screening for occult hepatitis B infection in low endemic, low socioeconomic settings.

Extensive Genetic Diversity of HIV-1 in Incident and Prevalent Infections among Malaysian Blood Donors: Multiple Introductions of HIV-1 Genotypes from Highly Prevalent Countries.

Transfusion-transmissible infections including HIV-1 continue to pose major risks for unsafe blood transfusions due to both window phase infections and divergent viruses that may not be detected by donor screening assays. Given the recent emergence of several HIV-1 circulating recombinant forms (CRFs) in high-risk populations in the Southeast Asia region, we investigated the genetic diversity of HIV-1 among the blood donors in Kuala Lumpur, Malaysia. A total of 211 HIV-positive plasma samples detected among 730,188 donations to the National Blood Centre between 2013 and 2014 were provided (90.5% male, median age: 27.0 years old). Recent or long-term infection status at the time of donation was determined using a limiting antigen avidity enzyme immunoassay (LAg-Avidity EIA). HIV-1 gag-pol genes were amplified and sequenced from residual plasma for 149 cases followed by genotype determination using phylogenetic and recombination analyses. Transmitted antiretroviral resistance mutations were not observed among the blood donors, among which 22.7% were classified as recent or incident infections. Major circulating HIV-1 genotypes determined by neighbour-joining phylogenetic inference included CRF01_AE at 40.9% (61/149), CRF33_01B at 21.5% (32/149), and subtype B at 10.1% (15/149). Newly-described CRFs including CRF54_01B circulated at 4.0%, CRF74_01B at 2.0%, and CRF53_01B and CRF48_01B at 0.7% each. Interestingly, unique HIV-1 genotypes including African subtype G (8.7%), CRF45_cpx (1.3%), CRF02_AG (0.7%) and CRF07_BC (0.7%) from China were detected for the first time in the country. A cluster of subtype G sequences formed a distinct founder sub-lineage within the African strains. In addition, 8.7% (13/149) of HIV-infected donors had unique recombinant forms (URFs) including CRF01_AE/B' (4.7%), B'/C (2.7%) and B'/G (1.3%) recombinants. Detailed analysis identified similar recombinant structures with shared parental strains among the B'/C and B'/G URFs, some of which were sequenced from recently infected individuals, indicating the possible emergence and on-going spread of foreign clades of CRF candidates among the local population. The findings demonstrate extensive molecular complexity of HIV-1 among the infected blood donors in Malaysia, driven in part by the increased spread of recently described CRFs and multiple introductions of previously unreported genotypes from highly prevalent countries.

Naturally occurring hepatitis B virus surface antigen mutant variants in Malaysian blood donors and vaccinees.

Hepatitis B virus surface mutants are of enormous importance because they are capable of escaping detection by serology and can infect both vaccinated and unvaccinated populations, thus putting the whole population at risk. This study aimed to detect and characterise hepatitis B-escaped mutants among blood donors and vaccinees. One thousand serum samples were collected for this study from blood donors and vaccinees. Hepatitis B surface antigen, antibodies and core antibodies were tested using a commercial enzyme-linked immunosorbent assay (ELISA) kit. DNA detection was performed via nested polymerase chain reaction (PCR), and the S gene was sequenced and analysed using bioinformatics. Of the 1,000 samples that were screened, 5.5 % (55/1,000) were found to be HBsAg-negative and anti-HBc- and HBV DNA-positive. All 55 isolates were found to belong to genotype B. Several mutations were found across all the sequences from synonymous and non-synonymous mutations, with the most nucleotide mutations occurring at position 342, where adenine was replaced by guanine, and cytosine at position 46 was replaced by adenine in 96.4 % and 98 % of the isolates, respectively. Mutation at position 16 of the amino acid sequence was found to be common to all the Malaysian isolates, with 85.7 % of the mutations occurring outside the major hydrophilic region. This study revealed a prevalence of 5.5 % for hepatitis B-escaped mutations among blood donors and vaccinated undergraduates, with the most common mutation being found at position 16, where glutamine was substituted with lysine.

WITHDRAWN: Rh genotypes among Malaysian blood donors

This article has been withdrawn at the request of the author and editor. The Publisher apologizes for any inconvenience this may cause. The full Elsevier Policy on Article Withdrawal can be found at http://www.elsevier.com/locate/withdrawalpolicy.

RH genotypes among Malaysian blood donors

Abstract Background: RH genotyping studies have been conducted mainly in people of Caucasian and African descent. There is limited information regarding the molecular basis for RH genotypes in Malaysia. Objectives: To investigate the prevalence and characteristics of RHCE genotypes among different ethnic groups in Malaysia. Methods: A total of 1014 whole blood samples were obtained from donors from 4 different ethnic groups (360 Malays, 434 Chinese, 164 Indians, and 56 others). All samples were phenotyped for C, c, D, E, and e using standard serologic methods and genotyped using polymerase chain reaction (PCR)-based analysis. Results: In the blood samples that we analyzed, the distribution of RH genotype antigens was significantly different among the various ethnic groups. Our findings showed that CCDee is the most common in Malaysian blood donors; 18.4% (187/1014) compared with other genotypes. The ccDEE genotype is more prevalent in the Chinese: 65.6% (82/125), and the ccee genotype is more prevalent in Indians: 47.1% (65/138). There were discrepancies between phenotypes and genotypes. There were 17 (1.7%) discrepancies in RH C/c genotyping results and of these 47% (8/17) occurred in Malays. Discrepancies in RH E/e results occurred in 3 samples (0.3%). Conclusions: Our study provides a database for the distribution of RH genotypes of donors from the major ethnic groups in Malaysia. Methods used in this study are useful for comparing the phenotypes and genotypes. Further investigation should be conducted to study the causes of these discrepancies using other molecular based techniques.

Evaluation of serological transfusion-transmitted viral diseases and mutliplex nucleic acid testing in malaysian blood donors

Transmission of infectious diseases is a recognized complication of blood transfusion and blood products. Nucleic acid testing (NAT) may contribute to improved efficiency of blood screening and thereby increase the safety margin for transfused blood. Unscreened blood samples from 1388 randomized donors were selected for this study at the Transfusion Medicine Unit of Hospital Universiti Sains Malaysia (HUSM). Informed consent was obtained from all donors and blood samples were tested for HIV, HBV and HCV serologically and by NAT assay. Of the 1,388 tested samples, 1,360 were non-reactive for both assays. Four samples (0.29%) were both serologically and NAT reactive. The remaining twenty-four samples were divided into two groups. Of these, five samples (0.366%) were NAT reactive and nineteen samples (1.37%) were serologically reactive. However, serology confirmation tests run on the latter nineteen samples were non-reactive. Hence, NAT adds benefit of detecting "false positive" reactions via standard serology, the cost of administering NAT also need further consideration and study.

Molecular analysis of Malaysian Chinese D- donors: a single centre experience

Background: The Rh blood group system is highly polymorphic and next to the ABO system is the most clinically significant in transfusion medicine. The frequency of D- phenotypes and the underlying molecular genetics vary widely in different populations. Objectives: We determined the prevalence of different D- phenotypes among Malaysian blood donors in a tertiary medical centre and identified the molecular basis of Chinese D- donors in this population. Materials and methods: A total of 146 D- Chinese donors with various Rh phenotypes were identified from review of blood donor records between January 2003 and September 2008. Fresh blood samples from 36 of these donors were obtained and further characterized by PCR-SSP to determine the molecular basis of these D- individuals. Results: A total of 86,620 blood donor records were reviewed. Of these 911 were D-, consisting of 483 Indians, 189 Malays and 146 Chinese. The ccee phenotype was the most common among D- individuals with a prevalence of 91.51% (442/483) in Indians, 74.60% (141/189) in Malays and 55.48% (81/146) in Chinese. D- phenotypes with C and/or E antigens were most common in Chinese {44.52% (65/146)}.In the molecular analysis of the 36 D- Chinese donor samples, 19 samples with ccee phenotype and 5/17 of samples with Ccee phenotype showed no detectable RHD gene. The remaining 12/17 Ccee samples had intact RHD genes with RHD (K409K) mutation. Conclusion: In our donor population, we found a wide variation in the incidence of D- as well as the distribution of various Dphenotypes among the three major ethnic groups. A significant number of D- Chinese donors with Ccee phenotype were found to be DEL with RHD (K409K) mutation. DEL red cells are known to cause anti-D alloimmunization. Therefore, in clinical practice, it is important to exclude DEL RBCs from D- donor pools. Keywords: Chinese D- donors, DEL variant, Malaysian blood donors, molecular genotyping, RhD-negative phenotypes

Diversity of Hepatitis B virus infecting Malaysian candidate blood donors is driven by viral and host factors

Malaysia is a medium endemic country for hepatitis B virus (HBV) infection but little is known about HBV strains circulating in Malaysian blood donors. Viral load, HBsAg concentrations and nested PCR products from 84 HBV surface antigen (HBsAg) positive samples were analysed in detail. Median viral load was 3050 IU/mL and median HBsAg 1150 IU/mL. Fifty-six full genome, 20 pre-S/S, 1 S gene and six basic core promoter/precore-only sequences were obtained. Genotypes B and C were present at a ratio of 2:1, and two genotype D samples were obtained, both from donors of Indian background. Phylogenetically, genotype B was more diverse with subgenotypes B2-5, B7 and B8 present, while most genotype C strains were from subgenotype C1. Genotypes B and C were equally frequent in ethnic Malays, but 80% of strains from Chinese were genotype B. HBsAg concentrations were higher in genotype C than in genotype B, in Chinese than Malays and in donors under the age of 30. HBV vaccine escape substitutions (P120S/T, I126N and G145G) were present in six strains. In the large surface protein, immuno-inactive regions were more mutated than CD8 epitopes and the major hydrophilic region. Strains of genotype B or from ethnic Malays had higher genetic diversity than strains of genotype C or from Chinese donors. Hence HBV strains circulating in Malaysia are phylogenetically diverse reflecting the ethnic mix of its population. Ethnic Malays carry lower HBsAg levels and higher genetic diversity of the surface antigen, possibly resulting in more effective immune control of the infection.

Detection of IgM Antibodies against &lt;i&gt;Legionella pneumophila&lt;/i&gt; Serogroup 1 in Malaysian Blood Donors and Patients with Respiratory Illnesses: Evaluation of Enzyme-Linked Immunosorbent Assay and Indirect Immunofluorescence Assay

Detection of IgM Antibodies against &lt;i&gt;Legionella pneumophila&lt;/i&gt; Serogroup 1 in Malaysian Blood Donors and Patients with Respiratory Illnesses: Evaluation of Enzyme-Linked Immunosorbent Assay and Indirect Immunofluorescence Assay

Identification of breakpoints in intergenotypic recombinants of HIV type 1 by bootscanning.

Identification of breakpoints in intergenotypic recombinants of HIV type 1 by bootscanning.

Assessment of infectiousness of male Malaysian blood donors.

HBeAg and anti-HBe were determined in the blood of 189 male blood donors. The incidence of HBsAg was 6.9% while that for HBeAg and anti-HBe was 1.6 and 18%, respectively. Of the 13 samples positive for HBsAg, two (15.4%) were positive for HBe while six (46.2%) were positive for anti-HBe. One specimen was negative for HBsAg but was positive for HBeAg and anti-HBe. The observations are discussed.

Assessment of Infectiousness of Male Malaysian Blood Donors

HBeAg and anti-HBe were determined in the blood of 189 male blood donors. The incidence of HBsAg was 6.9% while that for HBeAg and anti-HBe was 1.6 and 18%, respectively. Of the 13 samples positive for HBsAg, two (15.4%) were positive for HBe while six (46.2%) were positive for anti-HBe. One specimen was negative for HBsAg but was positive for HBeAg and anti-HBe. The observations are discussed.