Types Of Discrepancies Research Articles

Identification of a Rare Ael05/B101 Subtype and Selection of Blood Transfusion Strategy

To explore the mechanism of ABO discrepancy in a patient by ABO genotyping and the reasonable blood transfusion strategy. Routine serological test was carried out to analyze ABO blood group. The presence of the blood group determinants on the red blood cells were determined by adsorption-elution test. Exons 1-7 and adjacent introns of the ABO gene were amplified by PCR and sequenced. The patient showed ABO forward and reverse typing discrepancy. ABO forward typing defined as B, however, the reverse typing indicated that the patient was AB subtype. Absorption-elution test confirmed weak A antigens on the patient's red blood cells. The ABO gene sequencing showed a T>C exchange at position in exon 7 which resulted in a isoleucine to threonine substitution at codon 256. The ABO blood group genotype was ABO*Ael05/B101. The 767 T>C substitution in the gene of α-1,3-N-acetyl galactose is the molecular mechanism leading to the decrease expression of A antigen of the Ael05 subtype.

Gene identification of rare B(A) blood group

ObjectiveTo study serologic and gene characteristics of the B(A) blood group of blood donation volunteers in Jilin Province, China. MethodsABO subgroups were identified by standard serologic techniques in ABO typing discrepancy samples from all donors at the Jilin Blood Center (410,354 non-repeat donors). DNA (deoxyribonucleic acid) was collected from each sample and PCR (polymerase chain reaction) was used to sequence exons 6 and 7 and intron 6, part 5 from the ABO subgroup samples. PCR products were sequenced to identify ABO subgroups and the B(A) allele. ResultsFour cases of B(A) blood type were found after sequencing, including two different alleles: B(A)02 and B(A)04. Three of the four alleles were B(A)04. ConclusionAmong blood donation volunteers in Jilin Province, China, B(A)04 is the most common B(A) blood group allele, followed by B(A)02. The B(A) blood group is associated with a complicated serologic phenotype and DNA detection is necessary for this atypical phenotype sample.

On the prediction of X-ray dose deviations and the influence of CT scan protocols

Abstract Patients undergoing computerized tomography (CT) scans for tumor localization and treatment planning are frequently scanned using pre-set customized exposure protocols for optimal imaging of different anatomical sites. The question arises if these scanning protocols will produce a deviation in the Hounsfield number for a given tissue that can afterwards be used to predict the resulting dose calculation deviation due to this. The question is also if the deviation in the Hounsfield number of a tissue is large enough to affect dose calculation clinically significant. A study was devised in which a RMI phantom was scanned with five different scanning protocols and two CT beam energies at 120 and 135 kV. To assess the effect of insert configuration, Hounsfield number measurements were repeated for high density RMI inserts in the center and outer rings in the phantom. For each material insert the standard deviation of the Hounsfield number was calculated. To assist in dose prediction a series of DOSXYZnrc Monte Carlo calculations were carried out for beam qualities between 6 and 16 MV for a range of Hounsfield numbers calculated for bone and water. This provided information on how the depth dose varied as a function of Hounsfield number variation. Lastly, a series of treatment plans were setup for absorbed dose calculation using the RMI insert electron densities vs Hounsfield relations measured above. The absorbed dose of corresponding plans with the largest Hounsfield number variation were subtracted to find the dose discrepancies. It was found that the dose discrepancies in tissue types could be indicated by the deviation of the Hounsfield number due to different scanning protocols. The calculated dose difference were in all cases within 3%.

The potential association of the transcription levels of the ABO gene with the disease phases in AML patients

Patients with AML may show ABO blood typing discrepancy, and the expression levels of the ABO antigens may show some alterations with the disease progression. To better understand this phenomenon, the blood samples of 25 AML patients and 25 healthy blood donors were examined. The serological ABO blood types of the patients were determined in different AML stages, and gene sequencing was performed to identify the precise ABO genotypes. Reverse transcription-polymerase chain reaction (RT-PCR) was carried out to detect the transcription levels of the antigens. The genotyping result showed that there were 4 patients with genotype A1O, 5 patients with B1O, and 16 patients with A1B1. RT-PCR results indicated that the transcription levels of the ABO gene in 76% (19/25) of the patients were significantly lower compared with those in controls (p <0.05). After therapy, 3/4 patients with A1O returned to normal A, 4/5 patients with B1O returned to normal B, and 10/16 patients with A1B1 returned to normal AB. The patients who achieved complete remission (CR) showed no difference of transcription levels of the ABO gene from those of controls. The data indicated that the transcription levels of the ABO gene changed with the disease progression, suggesting its potential role in the progression of AML disease.

RHD alleles among pregnant women with serologic discrepant weak D phenotypes from a multiethnic population and risk of alloimmunization.

A considerable number of RHD alleles responsible for weak and partial D phenotypes have been identified. Serologic determination of these phenotypes is often doubtful and makes genetic analysis of RHD gene highly desirable in transfusion recipients and pregnant women. We analyzed the RHD gene in a cohort of pregnant women with doubtful D phenotypes. RHD genotyping was performed on 104 cases with D typing discrepancies or with history of serologic weak D phenotype. Laboratory-developed DNA tests, RHD BeadChip (Bioarray Solutions, Immucor), and sequencing were used to identify the RHD alleles. Molecular analyses showed 23 of 104 (22%) pregnant women were RHD*weak D types 1, 2, or 3 and not at risk for anti-D. Fifty-one (49%) were RHD*weak partial 4.0, 6 RHD*weak D type 38 (6%), 1 RHD*weak D type 45 (1%), 1 RHD*weak D type 67 (1%), and potentially at risk for being alloimmunized and making anti-D. Partial D was identified in 22 of 104 (21%) patients and definitively at risk for anti-D. Appropriate classification of RhD phenotypes is recommended for correct indication of RhIG in pregnant women. However, the serologic distinction between RhD-negative and RhD-positive phenotypes is a difficult task in the case of D variants due to the variations in serologic testing. Our results show a great variability in RHD variant alleles in pregnant women from this population of high admixture. According to these results, 78% of these obstetric patients are at risk for anti-D and candidates for RhIG.

Suicides in the Kyrgyz Republic: Discrepancies in different types of official statistics

In order to develop the “prevention of suicides” component of the state program on mental health protection of the Kyrgyz Republic population for 2017–2030 years, a suicide situation in the country over the past 10 years was investigated. During collecting, processing and statistical analysis of data from different sources, a significant discrepancy was revealed between them.According to the national statistics committee (NSC), in 2015 the level of prevalence of suicide was 6.93% per 100,000 of population, whereas, according to the information from the republican medical information center it was 4.82%. Over the last 10 years in the Kyrgyz Republic, a level of suicidal behavior among children, teenagers and young adults has still been rather high and 22.63% of suicide attempts were committed by persons aged between 18 and 22 years. A significant difference was revealed in the indexes of suicides among these age categories provided by NSC and the ministry of Inner affairs.Due to religious and national traditions, suicide is a very sensitive topic in Kyrgyzstan. In a lot of cases, death from suicide is not registered or is disguised as accidents and other causes of death, so the figures may be considered reliable.Thus, despite the relative standardization of suicides accounting by the separate departments, the further work is needed for coordination and harmonization of the data collection, as well as for development and implementation of inter-agency action plan to prevent suicides at the national level, taking into account the regional, cultural and ethnic characteristics.Disclosure of interestThe authors have not supplied their declaration of competing interest.

Systematic analysis of gene expression alterations and clinical outcomes of adenylate cyclase-associated protein in cancer.

Adenylate Cyclase-associated protein (CAP) is an evolutionarily conserved protein that regulates actin dynamics. Our previous study indicates that CAP1 is overexpressed in NSCLC tissues and correlated with poor clinical outcomes, but CAP1 in HeLa cells actually inhibited migration and invasion, the role of CAP was discrepancy in different cancer types. The present study aims to determine whether CAP can serve as a prognostic marker in human cancers. The CAP expression was assessed using Oncomine database to determine the gene alteration during carcinogenesis, the copy number alteration, or mutations of CAP using cBioPortal, International Cancer Genome Consortium, and Tumorscape database investigated, and the association between CAP expression and the survival of cancer patient using Kaplan-Meier plotter and PrognoScan database evaluated. Therefore, the functional correlation between CAP expression and cancer phenotypes can be established; wherein CAP might serve as a diagnostic marker or therapeutic target for certain types of cancers.

Comparative Analysis of Clinical Samples Showing Weak Serum Reaction on AutoVue System Causing ABO Blood Typing Discrepancies

BackgroundABO blood typing in pre-transfusion testing is a major component of the high workload in blood banks that therefore requires automation. We often experienced discrepant results from an automated system, especially weak serum reactions. We evaluated the discrepant results by the reference manual method to confirm ABO blood typing.MethodsIn total, 13,113 blood samples were tested with the AutoVue system; all samples were run in parallel with the reference manual method according to the laboratory protocol.ResultsThe AutoVue system confirmed ABO blood typing of 12,816 samples (97.7%), and these results were concordant with those of the manual method. The remaining 297 samples (2.3%) showed discrepant results in the AutoVue system and were confirmed by the manual method. The discrepant results involved weak serum reactions (<2+ reaction grade), extra serum reactions, samples from patients who had received stem cell transplants, ABO subgroups, and specific system error messages. Among the 98 samples showing ≤1+ reaction grade in the AutoVue system, 70 samples (71.4%) showed a normal serum reaction (≥2+ reaction grade) with the manual method, and 28 samples (28.6%) showed weak serum reaction in both methods.ConclusionsABO blood tying of 97.7% samples could be confirmed by the AutoVue system and a small proportion (2.3%) needed to be re-evaluated by the manual method. Samples with a 2+ reaction grade in serum typing do not need to be evaluated manually, while those with ≤1+ reaction grade do.

PERBEDAAN GOLONGAN DARAH ABO DI ANEMIA HEMOLITIK AUTOIMUN

Discrepancy of blood type is the difference result between cell grouping and serum grouping. The technical errors and various conditions are the common cause of their discrepancy, one of the conditions is mentioned as Auto Immune Haemolytic Anaemia (AIHA). Auto immune haemolytic anemia cold type can cause discrepancy due to cold autoantibody reaction. In this study the researchers conducted further testing to determine the actual blood type so the right blood transfusion can be given. A 41 years old woman with weakness and pale since a month before her admission to the present hospital, has been treated in a district hospital with the same condition. On the physical examination, the researchers found hepatomegaly. The complete blood count showed hemoglobin level was 2.1 g/dL and leukocyte count was 21.54í—103/uL. The clinical diagnosis made by the emergency department was anemia pro evaluation and planned blood transfusion with 500 cc of PRC. The peripheral blood smear shown hemolytic anemia. The result of the initial blood type test on forward grouping were AB positive agglutination and on reverse grouping O positive, then followed by washing erythrocyte and the researchers found the similar result. Because of the extra antibody found in both examination, the examination was carried out at 37°C of the patient's temperature. The researchers found that the blood type of AB rhesus positive with cold autoantibody. The indirect coombs test was positive. The patient is had been reported with ABO discrepancy cold type autoantibody (group IV) in autoimmune hemolytic anemia cold type. Further screening antibody tests are needed to confirm the specific antibody.

OR48 Resolution of conflicting hla assignment due to loss of heterozygosity in the hla region by NGS typing

57 year old female patient diagnosed with acute myeloid leukemia (AML) was referred to bone marrow transplant. Transplant workup and search for compatible unrelated donor was started immediately, along with induction therapy for AML. HLA typing performed from peripheral blood using high-resolution SSO method showed homozygous typing for all loci, except HLA C. However, the quality of the typing data, along with the unusual B and C allele association prompted confirmatory typing by next-generation sequencing (NGS), which revealed the presence of a previously undetected HLA B∗40:01:02 allele at 14%, and confirmed the presence of HLA C∗03:03:01 allele at 16%, consistent with a dramatic allelic imbalance. Research on the patient’s clinical history revealed that at the time of diagnosis and sampling, a significant population of atypical monocytic cells with promonocytes/monoblasts was detected by flow cytometry, accounting for at least 55% of the total leukocytes in the peripheral blood. Genomic microarray testing showed a 35.3 Mb terminal long contiguous stretch of homozygosity 6p25.3p21.31, which includes the HLA locus, resulting in copy-neutral loss of heterozygosity (LOH) acquired likely due to the malignancy. The LOH explains why one HLA haplotype was underrepresented in the initial blood sample acquired before the treatment. Buccal DNA sample was requested and tested with SSO method, which confirmed the presence of the alleles detected by NGS. In addition, confirmatory HLA typing performed 3 months later from peripheral blood post-treatment gave identical results. This supports the finding that the initial typing discrepancy was due to the overwhelming presence of malignant cells with LOH involving the HLA locus in the patient’s peripheral blood, collected before initiation of treatment. NGS typing using the Omixon Holotype platform was able to provide accurate typing of a sample with allelic disequilibrium, and allowed not only for detection but also for quantification of the minor alleles. This case highlights the importance of confirmatory typing pre-transplant for patients with hematological malignancies, since the presence of malignant cells in the peripheral blood may bias typing data if LOH involving the 6p region is present, which is a recurrent finding in patients with aplastic anemia or AML.

Organizational dynamic embeddedness and external shocks: The impact of financial and recession crises in strategic networks of the global pharmaceutical industry

We analyze the global pharmaceutical industry network using a unique database that covers strategic transactions (i.e., alliance, financing and acquisition collaborations) for the top 90 global pharmaceutical firms and their ego‐network partnerships totaling 4735 members during 1991–2012. The article explores insights on dynamic embeddedness analysis under network perturbations by exploring core and full networks' behavior during the global financial crisis of 2007–2008 and the subsequent global and Eurozone recessions of 2009–2012. We introduce and test literature grounded hypotheses as well as report network visualizations and nonparametric tests that reveal important discrepancies in both network types before and after the financial crisis offset. We observe that firms in core and full networks behave differently, with smaller top pharmaceutical firms of core networks particularly being affected by the crises, potentially due to a collaboration reduction with bigger top pharmaceuticals. On the other hand, big pharmaceuticals in full networks maintain their centrality position as a possible consequence of their strategic collaborations not only with other similarly sized firms but also due to their connections with subsidiaries and other private entities present in the total sample. Our results confirm the significant dynamicity reduction during financial crisis and recession periods for core and full networks, and highlight the importance that exogenous factors as well as network types play in centrality‐based dynamic longitudinal network analysis. © 2016 Wiley Periodicals, Inc. Complexity 21: 602–621, 2016

Variant RHD Types in Brazilians With Discrepancies in RhD Typing.

The knowledge of D variants in patients and donors is important because anti-D alloimmunization can occur in some but not all individuals who express a variant RHD allele. Serologic distinction of RhD discrepancies is not always straightforward, which makes molecular analysis highly desirable. A group of 223 subjects, 129 patients, and 94 blood donors was identified and analyzed on the basis of a D typing discrepancy. The D antigen expression was evaluated by tube and gel hemagglutination with four anti-D reagents. PCR-single specific primer (SSP), multiplex PCR, RHD BeadChip (Immucor), or sequencing were used for molecular analysis. In total, 168/223 (75%) weak D and 55/223 (25%) partial D variants were identified. Hemagglutination results varied in methods and anti-D reagents used in this process. There was no standard serologic reactivity identified, which could predict what type of D variant would be identified. Among weak D samples, types 1-3 were the most common, while DAR and DVI were most prevalent among partial D samples. Our results show that discrepancies found in the serologic typing should be investigated by molecular methods in order to determine the D variant involved and also to distinguish between weak D and partial D. The knowledge of the distribution of weak D types and partial D among populations is important for D- patients and pregnant women management.

Identification of new KLF1 and LU alleles during the resolution of Lutheran typing discrepancies.

The Lu(b) antigen is expressed on red blood cells (RBCs) of the majority of individuals in all populations. Its absence in transfused patients may lead to alloantibody production and mild-to-moderate transfusion reactions, and in pregnancies to mild hemolytic disease of the fetus and newborn. This report describes the results of discrepancy resolution between apparent LU*A/LU*B or LU*B/LU*B genotypes and apparent Lu(b-) or Lu(b+ weak) phenotypes in one Asian and 10 Caucasian blood donors. Whole blood samples were analyzed by molecular methods to resolve discrepancies between Lu(b-) phenotypes detected by serology and Lu(b+) phenotypes predicted by genotyping. RBC agglutination assays were performed with commercial and patient antisera by tube or gel column methods. Genotyping was performed on commercial arrays that target the LU*A/LU*B polymorphism at Position c.230. The discrepancies were resolved by sequencing of genomic DNA and in some cases by sequencing of cloned DNA fragments. Eleven new alleles with coding sequence variants were identified, seven in the KLF1 gene, which are presumed to act dominantly to silence LU expression, and four in the LU gene itself. The alleles are KLF1*114delC, KLF1*298T, KLF1*304C,484insC, KLF1*304C,1000del2, KLF1*621G, KLF1*948delC, KLF1*1040A,1045delT, LU*B(559T,711T,714T), LU*B(611A,638T), LU*B(1049del2ins3), and LU*B(1306T,1340T,1671T,1742T). Besides confirming common phenotypes and detecting rare antigen-negative phenotypes, the use of molecular methods in blood donor typing can prompt the identification of new alleles through discrepancy resolution.

D5S818 Typing Discrepancy Between PowerPlex®Fusion and Other STR Kits Including GlobalFiler®Caused by a One-base Deletion in 31 Nucleotides Upstream of the Repeat Region

Short tandem repeat (STR) typing is widely used in forensic investigation. When the same DNA sample is analyzed with different STR typing kits, a typing discrepancy is occasionally observed. In this study, we examined the cause of a typing discrepancy in a sample at D5S818 locus. This sample was designated as 10, 12 using Identifiler(®) , Identifiler(®) Plus, GlobalFiler(®) , PowerPlex(®) 16HS, and PowerPlex(®) 18D, but as 9.3, 12 using PowerPlex(®) Fusion. Sequencing results indicated that the shorter allele in the sample had a deletion (U31Tdel) at 31 nucleotides upstream of the repeat region (AGAT)10 . This deletion was located in the binding site of the published D5S818 forward primer in PowerPlex(®) 16 and was only 9 and 11 nucleotides downstream of our estimated 5' end position of D5S818 forward primer in GlobalFiler(®) and PowerPlex(®) 18D, respectively. We also examined the effect of primer length on the heterozygous peak balance in this sample.

Advances in Blood Typing.

The clinical importance of blood group antigens relates to their ability to evoke immune antibodies that are capable of causing hemolysis. The most important antigens for safe transfusion are ABO and D (Rh), and typing for these antigens is routinely performed for patients awaiting transfusion, prenatal patients, and blood donors. Typing for other blood group antigens, typically of the Kell, Duffy, Kidd, and MNS blood groups, is sometimes necessary, for patients who have, or are likely to develop antibodies to these antigens. The most commonly used typing method is serological typing, based on hemagglutination reactions against specific antisera. This method is generally reliable and practical for routine use, but it has certain drawbacks. In recent years, molecular typing has emerged as an alternative or supplemental typing method. It is based on detecting the polymorphisms and mutations that control the expression of blood group antigens, and using this information to predict the probable antigen type. Molecular typing methods are useful when traditional serological typing methods cannot be used, as when a patient has been transfused and the sample is contaminated with red blood cells from the transfused blood component. Moreover, molecular typing methods can precisely identify clinically significant variant antigens that cannot be distinguished by serological typing; this capability has been exploited for the resolution of typing discrepancies and shows promise for the improved transfusion management of patients with sickle cell anemia. Despite its advantages, molecular typing has certain limitations, and it should be used in conjunction with serological methods.

Non-Intravenous Sedatives and Analgesics for Procedural Sedation for Imaging Procedures in Pediatric Patients.

The purpose of this study was to describe the method of delivery, dosage regimens, and outcomes of sedatives administered by extravascular route for imaging procedures in children. Medline, Embase, International Pharmaceutical Abstracts, and Cochrane Database of Systematic Reviews were searched using keywords "child", "midazolam", "ketamine", dexmedetomidine", "fentanyl", "nitrous oxide", and "imaging." Articles evaluating the use of extravascular sedation in children for imaging procedures published in English between 1946 and March 2015 were included. Two authors independently screened each article for inclusion. Reports were excluded if they did not contain sufficient details on dosage regimens and outcomes. Twenty reports representing 1,412 patients ranging in age from 0.33 to 19 years of age were included for analysis. Due to discrepancies in doses and types of analyses, statistical analyses were not performed. Oral midazolam was the most common agent evaluated; other agents included intranasal (IN) ketamine, IN midazolam, IN fentanyl, IN and transmucosal dexmedetomidine, and N2O. Most agents were considered efficacious compared with placebo. Most agents showed efficacy for sedation during imaging when delivered through an extra-vascular route. Selection of agents should be based on onset time, duration, patient acceptability, recovery time, and adverse events. More robust studies are necessary to determine the optimal agent and route to utilize for imaging procedures when sedation is needed.

A novel GYPB-A-B hybrid gene responsible for Ss and MN typing discrepancies.

Transfusion support of S-s- patients is very challenging but can now be alleviated by genotyping mutations in the GYPB gene to predict their U- or U+(var) phenotype. However, the phenotype predicted by genotyping does not always correspond to the observed phenotype of red blood cells (RBCs), which requires further investigation to avoid such a typing discrepancy in the future. In this study, we elucidated the case of an S-s- female patient of African origin who was predicted to be S+s- by our genotyping platform. Long-range polymerase chain reaction (PCR) amplification and extended Sanger sequencing were required. The Ss typing discrepancy in the proband resulted from a converted GYPB allele that encodes neither S nor s due to the replacement of Exon B4 of GYPB by the homologous Exon A4 of GYPA. In this novel GYPB-A-B hybrid gene, the GYPA segment actually starts just downstream of Exon B2, causing a MN typing discrepancy too. While the proband's RBCs were M+N-, the genotyping predicted the M+N+ phenotype. The reported GYPB-A-B hybrid gene constitutes a limitation for the accurate prediction of the MN and Ss phenotypes by current genotyping methods. A PCR assay was therefore developed to detect its presence.

Effect of database profile variation on drug safety assessment: an analysis of spontaneous adverse event reports of Japanese cases.

BackgroundThe use of a statistical approach to analyze cumulative adverse event (AE) reports has been encouraged by regulatory authorities. However, data variations affect statistical analyses (eg, signal detection). Further, differences in regulations, social issues, and health care systems can cause variations in AE data. The present study examined similarities and differences between two publicly available databases, ie, the Japanese Adverse Drug Event Report (JADER) database and the US Food and Drug Administration Adverse Event Reporting System (FAERS), and how they affect signal detection.MethodsTwo AE data sources from 2010 were examined, ie, JADER cases (JP) and Japanese cases extracted from the FAERS (FAERS-JP). Three methods for signals of disproportionate reporting, ie, the reporting odds ratio, Bayesian confidence propagation neural network, and Gamma Poisson Shrinker (GPS), were used on drug-event combinations for three substances frequently recorded in both systems.ResultsThe two databases showed similar elements of AE reports, but no option was provided for a shareable case identifier. The average number of AEs per case was 1.6±1.3 (maximum 37) in the JP and 3.3±3.5 (maximum 62) in the FAERS-JP. Between 5% and 57% of all AEs were signaled by three quantitative methods for etanercept, infliximab, and paroxetine. Signals identified by GPS for the JP and FAERS-JP, as referenced by Japanese labeling, showed higher positive sensitivity than was expected.ConclusionThe FAERS-JP was different from the JADER. Signals derived from both datasets identified different results, but shared certain signals. Discrepancies in type of AEs, drugs reported, and average number of AEs per case were potential contributing factors. This study will help those concerned with pharmacovigilance better understand the use and pitfalls of using spontaneous AE data.

Morozov-type discrepancy principle for nonlinear ill-posed problems under η-condition

For proving the existence of a regularization parameter under a Morozov- type discrepancy principle for Tikhonov regularization of nonlinear ill-posed problems, it is required to impose additional nonlinearity assumptions on the forward operator. Lipschitz continuity of the Freechet derivative and requirement of the Lipschitz constant to depend on a source condition is one such restriction (Ramlau P, Numer. Funct. Anal. Optim. 23(1&22 )( 2003) 147-172). Another nonlinearity condition considered by Scherzer (Computing, 51 (1993) 45-60) was by requiring the forward operator to be close to a linear operator in a restricted sense. A seemingly natural nonlinear assump- tion which appears in many applications which attracted attention in various contexts of the study of nonlinear problems is the so-called η-condition. However, a Morozov-type discrepancy principle together with η-condition does not seem to have been studied, except in a recent paper by the author (Bull. Aust. Math. Soc. 79 (2009) 337-342), where error estimates under a general source condition is derived, by assuming the existence of the parameter. In this paper, the existence of the parameter satisfying a Morozov-type discrepancy principle is proved under the η-condition on the forward operator, by assuming the source condition as in the papers of Scherzer (Computing, 51 (1993) 45-60) and Ramlau (Numer. Funct. Anal. Optim. 23(1&22 )( 2003) 147-172). This source condition is, in fact, a special case of the source condition in the author's paper (Bull. Aust. Math. Soc. 79 (2009) 337-342).

LBP13: HLA MUTATIONS OBSERVED IN EBV-TRANSFORMED AND EXPANDED B-LYMPHOBLASTOID CELL LINES (BLCLS)

Aim Swabs created from BLCLs transformed from peripheral blood mononuclear cells (PBMCs) or expanded from existing BLCLs are used in the Be The Match Registry (BTMR) HLA QC program. The aim of this project was to document mutation as a cause of HLA typing discrepancies observed in 2 BLCLs. The first discrepancy arose in expanded BLCLs, sequenced in 2002 on a donor-derived BLCL. BLCL swabs were routinely typed multiple times using SSO and SBT methodologies as B∗49:01; however, in 2009, an SBT lab reported a potential new allele, B∗49:15, which was described in 2010 and confirmed in 2011 on the expanded BLCL by two labs. B∗49:15 differs from B∗49:01:01 by a single non-synonymous nucleotide polymorphism at NT 176 in exon 2, G/C. In 2013, a lab reported that the B49-specific amplicon showed a double peak, suggesting two alleles were present. Typing of other loci ruled out contamination. The second discrepancy arose in a BLCL transformed from patient-derived PBMC, originally typed as C∗02:09 using DNA extracted from PBMC in 2004. BLCL swabs repeatedly typed as C∗02:02 by SSO, SBT, and NGS. C∗02:09 differs from C∗02:02:01 by a synonymous nucleotide polymorphism at NT 315 in exon 2, A/G, and a non-synonymous polymorphism at NT 613 in exon 3, C/T. Methods PBMCs, fresh buccal swabs and expanded BLCL swabs were obtained for SBT and NGS of B∗49:15. Archived extracted DNA, PBMCs, failed cell culture, and transformed BLCL swabs were obtained for SBT of C∗02:09. Results Three labs confirmed B∗49:01:01 by SBT and NGS on archived PBMCs and newly obtained buccal swabs. NGS revealed the BLCL DNA contained both B∗49:15 and B∗49:01:01 in a 64/36 ratio and a 75/25 ratio in 2 samples. C∗02:09 was confirmed in archived extracted DNA used for the original allele description and in archived PBMC. SBT confirmed C∗02:02 on a failed cell culture from the PBMC to BLCL expansion and on additional transformed BLCL swabs. Conclusion It is likely these discrepancies arose as the result of mutations that occurred during the transformation of PBMC to BLCL and/or during the expansion of existing BLCLs. As a result of this work, B∗49:15 was deleted from the IMGT/HLA database in 6/2014. C∗02:09 was typed by 2 labs using DNA from the same cell, but should now be confirmed by SBT on a second subject. Purified DNA extracted from whole blood has begun to replace BLCL swabs in the BTMR QC program.