Comparison between a laser lancing device and lancet for capillary blood sampling, capillary blood hemoglobin measurement, and blood typing.

BACKGROUND: Blood donor screening includes tests using capillary blood, which is usually obtained by finger pricking using a lancet; however, the lancet has some shortcomings, such as skin puncture pain and needle stick injury. Recently, laser lancing devices for finger-prick sampling have been developed. We compared capillary blood Hb (cHb) levels and blood typing results obtained using a laser lancing device with those obtained using a lancet. STUDY DESIGN AND METHODS: cHb levels, blood typing results, and skin puncture pain scores were assessed in 191 participants. Finger-prick sampling was performed using LMT-1000 (LaMeditech, Seoul, Korea) and a lancet on the same finger on different hands. Paired venous Hb (vHb) levels were assessed in 103 participants using an automated hematology analyzer and compared with the cHb levels obtained using both lancing devices. RESULTS: The paired cHb results obtained with the laser lancing device and lancet showed a strong correlation (r = 0.927, p < .001) without any significant difference (p = .113) and a substantial agreement (κ = 0.654) for the identification of participants with a low Hb level (<12.5 g/dl). cHb levels were significantly higher than vHb levels with both lancing devices (mean differences: 0.27-0.43 g/dl). The results of blood typing using the laser lancing device showed 100% accuracy. Use of the laser lancing device showed significantly lower skin puncture pain scores (p < .001). CONCLUSION: Use of a laser lancing device for capillary Hb measurement and blood typing showed accurate results, with significantly reduced skin puncture pain. Laser lancing devices could be feasible for donor screening tests.

Cross-linked chitosan biofunctionalized paper-based microfluidic device towards long term stabilization of blood typing antibodies.

Long term stability of antibodies at room temperature is a major challenge in the commercialization of point-of-care devices for diagnostics. Since chitosan has been proven to be an excellent biofunctionalization material, the effects of four different biofunctionalization processes were studied to improve the room temperature stability of antibodies immobilized on chitosan modified paper-based microfluidic devices using blood typing antibodies as candidates. The devices used in this work have a flower-shaped design with 4 test zones at each corner. In three zones Anti-A, Anti-B, and Anti-D (Anti-Rh) antibodies are immobilized and the fouth zone represents the control (no antibodies) after biofunctionalization. The biofunctionalization of the paper devices was done with chitosan and chitosan cross-linked with sodium triphosphate pentabasic, glutaraldehyde, and sodium hydroxide. These devices were used for blood typing assays using real blood samples. A similar assay was also performed on unmodified (non-biofunctionalized) paper devices for comparison. Chitosan based biofunctionalized paper-devices showed better stability, up to 100 days as compared to 14 days on unmodified paper, at room temperature. Such biofunctionalized paper-based devices will be suitable for on-field and remote testing without any technical expertise and requirement for the cold chain.

A comparison of three column agglutination tests for red blood cell alloantibody identification.

OBJECTIVE: Commercial kits of column tests for pre-transfusion testing have progressively replaced conventional tube tests in most laboratories. Aim of this study was to compare three commercial test cell panels for the identification of irregular red blood cell (RBC) alloantibodies. Overall, 44 samples with a positive indirect antiglobulin test (IAT) by routine testing were used for comparison of following panels: Ortho RESOLVE® panelC (Ortho Clinical Diagnostics (OCD), Milan, Italy), ID-DiaPanel(-P) (Bio-Rad Laboratories, CA, USA) and Identisera Diana(P) (Grifols, Barcelona, Spain). Column agglutination techniques were used, with microtubes containing either microgel (Bio-Rad/Grifols) or glass bead microparticles (Ortho). RESULTS: Alloantibody identification was possible in 38 samples, of which identical identification was shown in 33 samples by all methods. The remaining samples showed differences between certain methods, with the gel card system being superior to the glass card system for analyzing stored samples Considering that not all samples were evaluated in all three methods, the concordance rate reached 100% between Bio-Rad and Grifols, 90.5% between Bio-Rad and OCD, 86.5% between OCD and Grifols and 90.5% between all methods. Although differences in sensitivities were seen for specific antibodies, the three methods showed comparable performance for the identification of RBC alloantibodies.

Enhanced fully cellulose based forward and reverse blood typing assay.

This study presents an enhanced paper-based analytical device (PAD) for forward and reverse group blood typing. The proposed PAD uses a novel methodology, which provides highly reliable results on a fully cellulose based device. The PAD was printed on different cellulose substrates. These substrates were made of different cellulose fibers (sisal and eucalyptus), different grammages, refining steps, and wet additive content. Best parameters were chosen to achieve high reliability on both forward and reverse blood typing. The substrates were patterned with five hydrophilic channels and two hydrophobic areas. For reverse blood typing, the hemoagglutination reaction took place on the hydrophobic surface of the paper before being transferred to the paper web, where together with the forward blood typing tests were all washed with saline solution to read the results by elution. This device allows direct read-out of results; the stains show were agglutination happens. Different blood types were in full agreement between the reverse and forward method and in agreement with traditional methods. The time and simplicity of this methodology confirmed its utility.

Updates on practical ABC blood compatibility testing in cats. / Neuigkeiten zur praktischen ABC-Blutgruppen-Bestimmung bei Katzen.

In feline practice, blood groups were considered unimportant until the 1980s. Since then much has been learned. The most important blood group system in cats is the AB (renamed here as ABC) blood group system consisting of blood types A, B and AB (better referred to as C). Type B cats have strong anti-A alloantibodies potentially leading to incompatibility reactions during A-B mismatched transfusions or neonatal isoerythrolysis (NI) in type A and C (AB) kittens born to type B queens. Acute hemolytic transfusion reactions as well as NI have been clinically well documented in cats. Immunological and genetic tests have been established and blood typing and crossmatching test kits have become commercially available. This review updates the current knowledge of these blood types, their genetics, associated incompatibility reactions, and different diagnostic tools for avoiding such reactions in clinical practice.

Performance and reliability of a benchtop automated instrument for transfusion testing: a comparative multicenter clinical study in the US population.

BACKGROUND: Heavy workload in hospital transfusion services and blood centers necessitates the implementation of automated platforms. We evaluated the performance of Erytra Eflexis (Diagnostic Grifols), a recently developed midsize automated instrument for pretransfusion testing, in comparison with a US Food and Drug Administration (FDA)-cleared device (Erytra). Reproducibility and repeatability of the results were also investigated. STUDY DESIGN AND METHODS: Studies were conducted using the same card technology and reagents at three US sites. Tests were performed on 9174 specimens from hospital patients (55.61%) and blood donors (43.39%). Evaluations included 18,413 ABO/D/reverse typing; 9084 Rh phenotypes, 4640 K phenotypes, 2052 antibody screenings, 1232 antibody identifications, 469 direct antiglobulin tests, 612 IgG crossmatches, and 700 ABO-compatibility crossmatches. A reference blood panel was also sent to each center, for a total of 3900 replicate tests. Concordance between results with the two instruments and performance among the different centers were statistically evaluated. RESULTS: Agreement between instruments was 99.84% for 37,202 test results, with 61 discrepancies (0.16%). Percentages of positive and negative agreement were 99.82% and 99.85%, respectively. No discrepancies were observed in 12,276 tests for direct ABO/D grouping. Discrepancies were observed during antibody identification (n = 19), antibody screening (n = 15), and reverse grouping (n = 10). Investigations of the discrepancies were resolved in favor of the study instrument in 55.73% of the cases. Erytra Eflexis obtained the expected results in the reproducibility analysis. CONCLUSION: This multicenter study demonstrates that Erytra Eflexis with its gel card technology and reagents is reliable and substantially equivalent to the FDA-cleared instrument used as the reference.

A new reliable test for crossmatching: microplate hydrogel immunoassay technology.

OBJECTIVES: To provide a novel assay to detect incomplete antibodies in the crossmatching test. BACKGROUND: There is a requirement in China that both major and minor crossmatch tests are required. Among all methods of crossmatching, the tube anti-human globulin test requires tedious washing steps and is time-consuming, whereas the microcolumn gel immunoassay anti-human globulin test is susceptible to sample quality. METHODS: The process of the microplate hydrogel immunoassay anti-human globulin test involves the use of our patented hydrogel chromatography medium and U-bottom microplates pre-coated with goat anti-human globulin (AHG). A mixture of red blood cells (RBCs) and serum is centrifuged through the hydrogel under precise conditions. In incompatible reactions, the sensitised RBCs are captured by the pre-coated AHG and form a layer over the bottom of the well, whereas in compatible reactions, the unbound RBCs form a button at the bottom of the well. The sensitivity of this new approach and the performance when testing old specimens were evaluated. RESULTS: This approach was more convenient and slightly more sensitive than the tube anti-human globulin test and was superior to the microcolumn gel immunoassay when testing old specimens. CONCLUSION: In general, this assay is suitable for the routine clinical use of crossmatching to detect incomplete antibodies.

Finger-Actuated Microfluidic Display for Smart Blood Typing.

Accurate blood typing is required before transfusion. A number of methods have been developed to improve blood typing, but these are not user-friendly. Here, we have developed a microfluidic smart blood-typing device operated by finger actuation. The blood-typing result is displayed by means of microfluidic channels with the letter and the symbol of the corresponding blood type. To facilitate the mixing of blood and reagents, the two sample inlets are connected to a single actuation chamber. According to the agglutination aspect in the mixture, the fluids are directed to both the microslit filter channels and bypass channels, or only to the bypass channels. The dimension of the microslit filter being clogged by the red blood cell aggregates was optimized to achieve reliable blood-typing results. The flow rate ratio between two channels in the absence of agglutination was subjected to numerical analysis. With this device, blood typing was successfully performed by seven button pushes using less than 10 µL of blood within 30 s.

Multicentre evaluation of Erytra Eflexis®, a benchtop fully automated analyser with a compact design for routine use in blood transfusion laboratory.

OBJECTIVES: Evaluation of the compact benchtop Erytra Eflexis® automated analyser was performed at three health centres representing a range of routine transfusion workload. BACKGROUND: Automation instruments with the simplicity and flexibility adequate for small- to mid-sized blood transfusion services are an unmet need. METHODS: Performance in pre-transfusion testing (2109 ABO/D, 382 Rh/K phenotype, 2001 antibody screening, 113 antibody identification, 151 DAT, 88 extended phenotype; 655 cross matching) in comparison to Erytra® as reference device was assessed. Throughput [time to first result (TTFR), final turn-around time (TAT), processing rate] was calculated; usability and adaptability in laboratory practice under routine and with emergency samples were surveyed. RESULTS: Agreement between systems was 99·8% (11/5499 test discrepancies, all due to weak/doubtful positive reactions). Erytra Eflexis produced six true positives (two Rh/D, two B positives, two screening), four false positives (three screening and one cross matching) and one false negative (screening). Processing of eight routine samples with the Erytra Eflexis for ABO/Rh(D) and screening took 34-38 min and 32-37 min, respectively, independent of the simultaneous processing of a STAT sample, whether or not the incubator for STAT was reserved. In this scenario, a STAT sample requested within 2 min after the routine load was processed in 14-26 min. Processing rate tended to stabilise and optimise in the larger workloads, particularly in ABO/Rh(D)/K cards (16·7, 18 and 19·5 results/h for 10, 15 and 24 specimens, respectively). CONCLUSION: Erytra Eflexis analyser was found to be reliable and suitable for pre-transfusion routine tests performed in a small-/medium-sized blood transfusion laboratory.

Method for ABO Blood Group Testing Using a General-Purpose Automated Biochemical Analyzer.

BACKGROUND: We investigated a high-throughput and high-precision forward ABO blood typing screening method that utilizes a general-purpose biochemical analyzer to perform direct red blood cell sampling. METHODS: The blood group antisera used were Ortho® BioClone® Anti-A Serum and Ortho® BioClone® Anti-B Se-rum. AFFIRMAGEN® Reagent Red Blood Cells (Ortho Clinical Diagnostics) were used for AB standard red blood cells. The general-purpose biochemical analyzer employed was the TBATM-120FR HbA1c measurement unit (Canon Medical Systems). RESULTS: ABO blood group of patient samples was determined based on values relative to amount of change in the AFFIRMAGEN® response. Repeatability was CV5% or lower, and testing of 1,112 patient samples showed 100% agreement between the results obtained using the proposed method and those obtained using the tube test method. CONCLUSIONS: The proposed method allows ABO blood typing to be performed simply, quickly, and with a high degree of precision.

Chamfer-Type Capillary Stop Valve and Its Microfluidic Application to Blood Typing Tests.

This paper presents a novel design of a capillary stop valve with a chamfered side that can be used as a flow regulator to hold an injected microfluid in the valve position in a capillary force-driven microfluidic device. Biochemical analysis can be conducted if the chamfer-type valves are placed at strategic positions according to the test protocol. Hence, the stored reagent can be dragged out of the valve for further reaction when the specimen passes through. However, countercurrent phenomena were observed in the commonly used T-type capillary stop valve (without the chamfered side). In blood typing tests, the countercurrent led to incomplete dragging and the fluid stopped flowing at the complicated mixing channel; thus, the blood typing reaction was attenuated. On the contrary, the chamfer-type valve reduced the countercurrent phenomena and ameliorated the blood typing reaction. Consequently, agglutination results can be easily discriminated from nonagglutination cases.

Development of RBC Membrane Antigen Arrays for Validating Blood Grouping Reagents.

Antibody reagents have been remained as a standard approach to characterize blood group (BG) antigens in clinic. The specificity and cross-reactivity of these BG antibodies are routine detected using the gel microcolumn assay (GMA). However, the GMA is neither specific nor sensitive, thus increasing the risk of improperly matched RBC transfusions. In this work, we describe a bead-based RBC membrane antigen array to detect BG antibody-antigen binding with ∼700-fold higher sensitivity and dynamic range than the GMA. RBC membrane antigen arrays were fabricated using fragmented RBC membranes highly enriched in BG panel antigens. The arrays were then used to screen the interactions of 15 BG reagents to three antigen panels. The majority of the antibody reactions (i.e., 86.7%; 39/45) aligned with those obtained with the GMA. The six cross-reactive, nonspecific antibody reactions identified only by our arrays (i.e., 13.3%; 6/45) were confirmed by agglutination inhibition and genotyping assays. These results demonstrate that our RBC membrane antigen array has great potential in screening BG antibodies and improving the safety of RBC transfusions.

Medical Devices; Hematology and Pathology Devices; Classification of Blood Establishment Computer Software and Accessories. Final rule.

The Food and Drug Administration (FDA, Agency, or we) is issuing a final rule to classify blood establishment computer software (BECS) and BECS accessories (regulated under product code MMH) into class II (special controls). FDA has identified special controls for BECS and BECS accessories that are necessary to provide a reasonable assurance of safety and effectiveness. FDA is also giving notice that the Agency does not intend to exempt BECS and BECS accessories from premarket notification requirements of the Federal Food, Drug, and Cosmetic Act (FD&C Act).

Finger-actuated microfluidic device for the blood cross-matching test.

A blood cross-matching test should be carried out to prevent a hemolytic transfusion reaction as the final verification step. To simplify complicated procedures of a conventional blood cross-matching test requiring bulky systems and skilled people, we present a finger-actuated microfluidic device for the blood cross-matching test. Although finger actuation is a simple action that anyone can easily accomplish, there would be a variation in the individual finger actuation that may induce the user-dependent errors of the device. Therefore, the working principle of the finger-actuated microfluidic device is newly designed to reduce the user-dependent errors by indirectly controlling the pressure of fluidic channels. The constant volume was repeatedly dispensed by pushing and releasing a pressure chamber regardless of the different pushed depths of the pressure chamber, the pushing time interval, and the end-users. The dispensed volume was linearly increased according to the number of pushing times applied to the pressure chamber and determined by adjusting the diameter of an actuation chamber. In addition, multiple fluids can be dispensed with a desirable ratio by pushing and releasing the pressure chamber. Finally, a finger-actuated microfluidic device for the blood cross-matching test was developed, which can simultaneously actuate four fluidic channels. After loading 50 µL of whole blood samples from a donor and a recipient into two inlets of the device, the blood plasma from each individual was separated through the two plasma separation membranes. The blood cross-matching test results can be achieved by cross-reacting the donor's blood plasma with the recipient's whole blood as well as the donor's whole blood with the recipient's blood plasma by pushing and releasing only a single pressure chamber within 10 min.

Performance evaluation study of ID CORE XT, a high throughput blood group genotyping platform.

BACKGROUND: Traditionally, red blood cell antigens have been identified using serological methods, but recent advances in molecular biology have made the implementation of methods for genetic testing of most blood group antigens possible. The goal of this study was to validate the performance of the ID CORE XT blood group typing assay. MATERIALS AND METHODS: One thousand independent samples from donors, patients and neonates were collected from three research institutes in Spain and the Netherlands. DNA was extracted from EDTA-anticoagulated blood. The data were processed with the ID CORE XT to obtain the genotypes and the predicted blood group phenotypes, and results were compared to those obtained with well-established serological and molecular methods. All 1,000 samples were typed for major blood group antigens (C, c, E, e, K) and 371-830 samples were typed for other antigens depending on the rarity and availability of serology comparators. RESULTS: The incorrect call rate was 0%. Four "no calls" (rate: 0.014%) were resolved after repetition. The sensitivity of ID CORE XT for all phenotypes was 100% regarding serology. There was one discrepancy in E- antigen and 33 discrepancies in Fyb- antigen. After bidirectional sequencing, all discrepancies were resolved in favour of ID CORE XT (100% specificity). ID CORE XT detected infrequent antigens of Caucasians in the sample as well as rare allelic variants. DISCUSSION: In this evaluation performed in an extensive sample following the European Directive, the ID CORE XT blood genotyping assay performed as a reliable and accurate method for correctly predicting the genotype and phenotype of clinically relevant blood group antigens.

An Automatic Lab-on-Disc System for Blood Typing.

A blood-typing assay is a critical test to ensure the serological compatibility of a donor and an intended recipient prior to a blood transfusion. This article presents a lab-on-disc blood-typing system to conduct a total of eight assays for a patient, including forward-typing tests, reverse-typing tests, and irregular-antibody tests. These assays are carried out in a microfluidic disc simultaneously. A blood-typing apparatus was designed to automatically manipulate the disc. The blood type can be determined by integrating the results of red blood cell (RBC) agglutination in the microchannels. The experimental results of our current 40 blood samples show that the results agree with those examined in the hospital. The accuracy reaches 97.5%.

Evaluation of a lateral flow-based technology card for blood typing using a simplified protocol in a model of extreme blood sampling conditions.

BACKGROUND: Life-threatening situations requiring blood transfusion under extreme conditions or in remote and austere locations, such as the battlefield or in traffic accidents, would benefit from reliable blood typing practices that are easily understood by a nonscientist or nonlaboratory technician and provide quick results. STUDY DESIGN AND METHODS: A simplified protocol was developed for the lateral flow-based device MDmulticard ABO-D-Rh subgroups-K. Its performance was compared to a reference method (PK7300, Beckman Coulter) in native blood samples from donors. The method was tested on blood samples stressed in vitro as a model of hemorrhage cases (through hemodilution using physiologic serum) and dehydration (through hemoconcentration by removing an aliquot of plasma after centrifugation), respectively. RESULTS: A total of 146 tests were performed on 52 samples; 126 in the hemodilution group (42 for each native, diluted 1/2, and diluted 1/4 samples) and 20 in the hemoconcentration group (10 for each native and 10% concentrated samples). Hematocrit in the tested samples ranged from 9.8% to 57.6% while hemoglobin levels ranged from 3.2 to 20.1 g/dL. The phenotype profile detected with the MDmulticard using the simplified protocol resulted in 22 A, seven B, 20 O, and three AB, of which nine were D- and five were Kell positive. No discrepancies were found with respect to the results obtained with the reference method. CONCLUSION: The simplified protocol for MDmulticard use could be considered a reliable method for blood typing in extreme environment or emergency situations, worsened by red blood cell dilution or concentration.

A robust, portable and backflow-free micromixing device based on both capillary- and vacuum-driven flows.

In capillary- or vacuum-driven microfluidics, surge backflow events are common when merging or pumping two similar or dissimilar liquids together if a pressure difference exists between them. In this work, a robust, portable micromixing device that is insensitive to backflow was designed, fabricated and characterised. A capillary-driven pressure balancing bypass connected between two inlet ports diminished the initial pressure difference caused by capillarity and gravity present in each liquid at the two inlet ports. Then, using manual syringe-assisted vacuum-driven pumping that operated based on the high gas permeability of polydimethylsiloxane, the two pre-balanced liquid streams could synchronously enter a dead-end micromixing channel without any backflow. To test the performance of this device, we first used it to mix two aqueous solutions of different coloured dyes. We varied the initial volume difference between the solutions to study the effect of gravity-induced pressure difference on mixing. Next, as a proof-of-concept application, ABO/Rh blood groups were successfully determined through detection of blood antigen-antibody agglutination. The filling time of agglutinated samples, driven by the simple syringe-assisted pumping, in the dead-end mixing channel was consistently 10% longer than that of blood samples without the agglutination reaction. Thus, the proposed device shows great potential for use in a wide variety of blood typing assays, agglutination-based assays and point-of-care or lab-on-a-chip testing applications.

Analysis of ABO chimera from peripheral red cells and reticulocytes by flow cytometry and micro gel column technique in patients post-ABO-incompatible HSCT.

BACKGROUND: How to choose an appropriate method to monitoring dynamic ABO chimera post-ABO-incompatible HSCT is crucial to not only assess the status of erythroid engraftment but also achieve personalized safety transfusion. METHODS: We evaluated the efficacy of micro gel column technique (MGCT) and flow cytometry (FCM) by series of artificial ABO chimera mixtures from 0.5% to 50% and by investigating 15 cases of ABO-incompatible HSCT patients with longitudinally ABO blood grouping and ABO chimera from RBCs and reticulocytes. RESULTS: 5% and 2% of ABO chimera mixtures can be efficiently detected by MGCT and FCM, respectively. 6.3% of donor RBCs with 44.77% of donor reticulocytes in the early phase and 7.9% of patient RBCs with 96% of donor reticulocytes in the phase of complete donor type can be detected by FCM rather than failure by MGCT simultaneously. However, in case 8#, 8.6% of donor reticulocytes rather than 99.1% of donor RBCs on the 98th day post-HSCT could adequately predict early relapse. CONCLUSIONS: Investigation of ABO chimera from reticulocytes by FCM is a more effective strategy rather than ABO blood grouping and RBCs by FCM to indicate the true progress of erythroid alteration and achieve personalized safety transfusion post-ABO-incompatible HSCT.

A uniform method for the simultaneous blood group phenotyping of Fya , Fyb , Jka , Jkb , S, sÌ , P1, k applying lateral-flow technique.

BACKGROUND AND OBJECTIVES: A lateral flow assay for simultaneous blood group typing of ABO, RhD, C, E, c, e, Cw and K with stable end-point and without centrifugation is in routine use since several years (MDmulticard® ). The typing of extended phenotype parameters belonging to the Duffy, Kidd, MNSs blood group systems and others, however, has not yet been demonstrated for this technique. Reliable detection of Fyx , a weak Fyb phenotype with a pronounced quantitative reduction of the number of Fyb antigens on the erythrocyte surface, remains a weakness of current serological blood grouping techniques. MATERIAL AND METHODS: The performance characteristics of the following reagents were evaluated in donor and patient samples in lateral flow technology (MDmulticard® ): Anti-Fya , -Fyb , -Jka , -Jkb , -S, -s̅, -P1 and -k. The sensitivity to detect Fyx was in addition evaluated with Fyx positive samples, which had been preselected by MALDI-TOF MS-based genotyping. RESULTS: All results obtained with the MDmulticard® were in full accordance with those of the CE-certified reference products for all the eight reagent formulations used: Anti-Fya , -Fyb , -Jka , -Jkb , -S, -s̅, -P1 and -k. Also, all Fyx phenotypes of the selected population of 93 positive samples, originally identified by MALDI-TOF MS-based genotyping, were reliably detected by the lateral flow assay. CONCLUSION: Extended phenotype blood group parameters, including the serologically challenging Fyx phenotype, can be determined simultaneously, rapidly and accurately using the lateral flow (MDmulticard® ) technology, even in cases when IgG class antibodies are the only source of diagnostic antibodies.