Barcode-integrated cellulose based microfluidic system for intelligent point-of-care blood typing.

Karl landsteiner and his major contributions to haematology

Blood group serology

Implications of ABO Error Rates in Proficiency Testing for Solid Organ Transplantation

Blood group systems

Blood typing is a critical element of medical diagnostics and health assessment. Nevertheless, prevailing point-of-care testing (POCT) assays, including the slide method, tube method, column agglutination method, and gel test method, often necessitate costly equipment such as centrifuges, cumbersome procedures, and skilled personnel for operation. Furthermore, the testing time is relatively long (∼30 min), which is not applicable in emergency cases/point-of-care testing. Thus, a low-cost, simple, low blood consumption and rapid blood typing method is urgently required, especially in resource limited areas for point-of-care applications. Herein, we present a novel method of laser-based identification of red blood cell (RBC) agglutination for accurate and reliable blood typing. Specifically, the RBC agglutination reaction (a bio-signal) is initially converted into a laser signal. Subsequently, a photoresistor converts the laser signal into an electrical signal (with high or low resistance). Finally, the electrical signal is converted into an optical signal through a detection circuit (if the resistance is lower than 600 Ω, the LED light is on; otherwise, the LED light is off). By merely interpreting the light signals, even non-professionals can precisely determine the blood types without the risk of misinterpretation. This laser-based blood typing method can effectively avoid human errors caused by naked-eye observations or environmental interference and can provide new insights for developing accurate and reliable hemagglutination identification methods for point-of-care blood typing.

ABO blood grouping is an important antigenic blood typing tools in blood transfusion and organ transplants. Mismatching of blood during transfusion would lead to undesired transfusion reactions. Due to rare occurrence of rare blood group such as A2 subtype, regular blood grouping technique would have missed the identification of blood group. In this study, the identification of A2 subgroup using routine serological technique was validated via DNA sequencing technique. A total of 656 students participated in this study consist of Malay (87.0 %), Chinese (0.4 %), Indian (11.4 %) and others ethnic group (0.9%) respectively. Monoclonal antisera A, B, AB, D, A1 lectin and H lectin were used to identify the antigen on red blood cells. DNA sequence analysis was applied to examine single nucleotide polymorphisms (SNPs) at position 467 (substitution of C>T) and 1061 (deletion of C) on coding region of ABO gene. Our findings showed of 656 blood samples, 256 (39.0%) were blood group O, 190 (29.0%) were blood group B, 179 (27.3%) were blood group A and 31(4.7%) were blood group AB. The frequency of A1 subgroup is 177 (99.0%) and A2 subgroup is 2 (1.0%). From 179 A blood group, only 2 samples showed negative reaction towards anti-A1 lectin. DNA sequence analysis revealed the SNPs at nucleotide 1061 position in sample 2, however sample 1 did not have this mutation and the subgroup was not identified. DNA sequencing provides a precise and high accuracy in identification of A subgroups.

In recent years, paper-based diagnostic assays for blood typing have advanced rapidly for the typing of ABO and Rh(D) blood groups. Most of these paper-based assays were based on the direct agglutination mechanism using immunoglobulin M (IgM) antibodies treated on paper. The attempt to directly agglutinate red blood cells (RBCs) on paper using IgG antibodies has consistently failed. This was expected due to the ‘non- agglutinating’ nature of IgG antibodies. The indirect antiglobulin test (IAT), which is a 2- stage test including the sensitisation of red blood cells (RBCs) with specific IgG antibodies and the bridging of IgG-sensitised RBCs with anti-IgG, was developed to detect IgG antibodies and IgG-sensitised RBCs. The IAT is routinely used in RBC antigen phenotyping, antibody detection and identification, and cross-matching donor and recipient for blood transfusion. Standard methods for the IAT such as the traditional tube test, gel card column agglutination technology and solid phase assay are cumbersome, time consuming and requires modern laboratory equipment handled by skilled personnel. An easy-to-use paper diagnostic for IAT would be invaluable for any blood analysis involving the IgG antibodies. IgG antibodies can cross the human placenta, potentially resulting in haemolytic disease of the fetus and newborn; their detection is crucial. In this thesis, two paper-based IAT assays were developed as proofs of concept. Both assays involved the treatment of paper with anti-IgG that could capture IgG-sensitised RBCs. The first assay utilised the filtration ability of paper to trap agglutinated RBCs within the fibre network while allowing non-agglutinated RBCs to be filtered out through the pores between fibres. Instead, the second assay was based on the principle of paper chromatography. Once again, RBC agglutinates are trapped between the fibre networks, but non-agglutinated RBCs are eluted away from the sample loading point with the capillary force of a buffer solution wicking the inter-fibril capillaries of paper. These assays do not require specific laboratory equipment nor skills. They are simple to use and results are easy to read, eliminating human error. Given the different mechanisms on which both assays rely upon, the different needs and testing conditions required by end-users can be suited by selecting the proper test. Whilst the blood typing principle and current technologies are well established, the fundamentals of antibody – antigen interactions occurring within a blood typing system are poorly quantified. Blood typing specific antibody – antigen interactions have always been reported as affinity constants and relative binding units. The interaction force and energies have never been reported. In particular, the interactions between the anti-IgG and RBC surface antigen bound IgG antibodies have never been rigorously quantified. RBC surface antigen distribution have always been observed and indirectly evaluated using labelled antibodies coupled with scanning electron microscopy or flow cytometry techniques. These techniques involve over-manipulation of the RBCs and results are unspecific and general. The atomic force microscope (AFM) is a versatile instrument for biological imaging which can resolve features from the micro scale down to the nano scale. It also allows imaging not only in air but also in liquid, enabling imaging of biomolecules in physiological conditions. The unique force mapping mode of the AFM allows the simultaneous measurement of interaction forces and localization of sites where specific interactions have occurred. Using the AFM force mapping mode, we have quantified for the first time the interaction energies between anti-IgG – IgG on RBC and IgM – RBC surface antigen while simultaneously mapping the location and quantifying the density of the antigen sites. This was achieved by functionalizing the AFM cantilever tip with anti-IgG and IgM antibodies, respectively, in two separate studies. The antigen of interest in both studies was kept as the D antigen. For the first time, the distribution of antigens on individual RBCs as mapped by anti-IgG and the IgM-functionalized AFM tips was measured and compared under physiological conditions and found to be similar. However, heterogeneity observed in the distribution of antigens among RBCs isolated from a single donor and on RBCs isolated from multiple donors bearing the same blood group emphasized the importance of mapping antigen distribution on single cells rather than relying on an ensemble set of information. This technique has allowed the investigation of antigen distribution in real-time, in situ and label free. It is anticipated that the developed technique can be expanded to the use in the development of selective and personalized medical treatment.

Blood typing is an important medical immunodiagnostic test for human blood transfusion and organ transplantation. Today, the global demand for accurate and rapid blood typing diagnosis is extremely large. However, highly sophisticated blood typing techniques are not appropriate for resource-limited regions, as they are either too expensive or require professional technicians to operate them. There is also an extensive demand for low-cost blood typing techniques with the capabilities of automation and high-throughput operation in blood bank laboratories and hospitals. To solve these problems, the research reported in this thesis focuses on the investigation and development of blood typing devices based on bioactive paper, and liquid micro reactors fabricated using superhydrophobic materials. This thesis includes two parts, which present research work on blood typing techniques based on bioactive paper devices and liquid micro reactors. In the first part, research into the fundamental mechanisms of paper-based blood typing devices combines scientific information with microscopic techniques and expertise in papermaking. The agglutination and immobilization mechanisms of red blood cells (RBCs) in antibody-treated paper are explored using confocal microscopy. The transport pathways of RBCs within the fibre network of paper are studied using a combined dual beam system with scanning electron microscopy (SEM) and focused ion beam (FIB) technology. Both these microscopic methods developed are powerful techniques for providing the details of RBCs at cellular level inside paper. This part also demonstrates a potential application for controlling the performance of paper-based blood analysis devices through paper structure design, which can be achieved during the papermaking process. Clear understanding of the fundamental mechanisms is essential for the design and production of paper-based blood typing devices which meet the ASSURED criteria (affordable, sensitive, specific, user-friendly, rapid, equipment-free and deliverable to those who need them). It is also important for the development of other paper-based blood analysis devices, and will contribute to the improvement of public health situations in rural areas and developing countries. The second part of this thesis presents innovations in the application of liquid micro reactors fabricated from superhydrophobic materials for blood typing devices. Two design concepts for the production of liquid micro reactors are demonstrated in this part: one is a superhydrophobic surface-supported liquid drop; the other is a liquid marble – a liquid drop wrapped in superhydrophobic powder. In both cases, the near-spherical shape of the liquid micro reactors enables the devices to provide clear and magnified side views to facilitate the observation of the detailed processes of RBC haemagglutination. Most importantly, by integrating these devices with advanced image capture and processing techniques, the automation of high-throughput blood typing, including rapid assay result interpretation, data storage, and transmission, can be achieved. The author sincerely hopes that the findings presented in this thesis on the applications of bioactive paper and liquid micro reactors in blood typing will serve the community by providing high-performance, simple blood typing devices. The author also hopes that the findings will be extended to the development of other blood-based diagnostic devices.

BCSH guideline for the use of anti-D immunoglobulin for the prevention of haemolytic disease of the fetus and newborn.

Blood group detection is a crucial aspect in healthcare industry, especially during blood transfusions, organ transplantation, and prenatal care. Traditionally, blood typing has been achieved through serological tests. These are highly accurate tests but require a blood sample and are usually carried out in a laboratory. It can be invasive, take a lot of time, and relies on specialized equipment; hence, they are not easily accessible, especially to remote or resource-limited areas. This paper introduces a new approach for blood group detection through fingerprint image processing. Instead of depending on a blood sample we take, we look to utilize the uniqueness in our fingerprints — that is, ridge patterns and minutiae points — for determination of our blood type. Fingerprints are known to possess unique features that have, upon careful analysis, indicated a possible link to blood group traits. This method, using advanced image processing techniques and machine learning algorithms, particularly Convolutional Neural Networks (CNNs), can analyze fingerprint images and predict blood types with accuracy. It could transform the way blood typing is done by offering a non- invasive, quick, and affordable alternative that could be used in places where traditional blood typing is very challenging. The preliminary data look encouraging, indicating the potential of this approach for revolutionizing point-of-care diagnostics and making blood typing easier and more efficient and quick..

ABO and Rh blood group systems are the most important blood grouping systems from clinical aspect. Determination of blood group is important for blood transfusion therapy, medico-legal purposes, organ transplantation, settlement of paternity disputes etc. A cross-sectional descriptive study was carried out for a period of one year from 1st January 2011 to 31st December 2011 in blood bank of Tribhuvan University Teaching Hospital. All blood samples collected for blood group determination were included in the study. Blood group was determined by slide agglutination method using commercial antisera. A total of 13568 blood samples were analyzed, 5123 (37.75%) were male and 8445 (62.25%) were female. Frequencies of blood groups A, B, AB and O were found to be 4034 (29.7%), 3665 (27.0%), 1114 (8.2%) and 4755 (35.1%). Frequencies of Rh positive and Rh negative blood groups were found to be 13200 (97.3%) and 368 (2.7%). Blood group O was common in Brahmin, Chhetri, Tamang, Lama, Gurung, Sherpa, Terai Brahmin, Muslim and Yadav ethnicities; blood group A was common in Newar, Rai, Magar, Limbu and Sanyasi ethnicitites; and blood group B was common in Tharu and Marwari ethnicities. Blood group O was found to be the most common blood group while AB was the rarest one. It was found that blood group O is the more common in Sherpa, Brahmin and Yadav; A in Limbu, Rai and Newar; and B in Tharu and Marwari ethnicities.

Impact of Blood Group Type on Severity of Disease in COVID-19 Patients

Background Blood group, Bleeding Time and Clotting time are clinically useful tests, extensively used during blood transfusion, platelet disorders and a variety of forms of treatment in hospitals. The objective of our study was to assess the country wise distribution of blood groups and to compare the country wise bleeding time and clotting time with respect to blood group. Material and Methods This Observational Study has been performed at Manipal College of Medical Sciences, during the period of 1st May 2010 to 31st August–2011. The study was carried out on the undergraduate medical students of Basic sciences and included assessment of Bleeding time, Clotting time and Blood grouping by standard procedures. Results 261 medical students participated in the study. Among the Nepalese students, 30.5% had blood group A, 28.9% B, 5.5% AB and remaining 35.2% O. among the Indian students, 22.8% were blood group A, 45.7% B, 7.6% AB and 23.9% O. In Sri Lankan students, 22% had blood group A, 19.5% B, 4.9% AB, and 53.7% blood group O. Among Nepalese students, BT was higher in blood group A (163.85 seconds) as compared to Indians (154.29Sec) and Sri Lankans (133.33 sec). Nepalese students with AB blood group had higher BT (171.43 sec). Indian students with blood group A had comparatively higher clotting time (328.57 sec) followed by Nepali (276.15 sec) and then Sri Lankan students (270 sec). Girls are 4.432 times more prone for bleeding time greater than 4 minutes and 2.453 times for clotting time value above 6 minutes as compared to males. Conclusion Our study suggests that O blood group is predominant in Nepalese students while Blood group B was the most common blood group in the Indian students. We found higher BT and CT in females, which are more prominent in Indians, so this gender disparity is an additional risk factor for them. Health Ministries of respective countries have to improve the proper health care policies required for prevention and management of blood group diseases, bleeding time and clotting time related disorders like Hypoprothrombinemia and Thrombocytopenia.http://dx.doi.org/10.3126/nje.v1i4.5755 Nepal Journal of Epidemiology 2011;1(4):135-140

The ABO and Rh blood group systems are fundamental to the fields of transfusion medicine and blood banking, with the ABO system's discovery by Karl Landsteiner in 1900 marking a significant milestone in these domains. The distribution of ABO and Rh blood groups significantly varies across different populations and is crucial for effective blood bank management and transfusion services. A cross-sectional study was conducted on first year MBBS medical students in the Dinajpur Medical College, Dinajpur, Bangladesh from January to December 2021. Total of 120 medical students, 40 males and 80 females, samples were collected by finger prick method under aseptic precautions. The ABO blood grouping and Rhesus factors (Rh) typing determined by glass slide method, which is based on antigen antibody agglutination. ABO blood group system classifies blood groups of people into four different types namely A, B, O and AB. The need for the study of frequency distribution of blood group is multipurpose. Out of total 120 participants 80 were female students and 40 were male students. Among the most common blood group was O (42.5%) followed by B (29.2%), A (16.6%), AB (11.6%). 80.8 % students were Rh positive and 19.2 % were Rh negative and O + is (30.8%) commonest blood group among 120 students and A- and AB – are rarest blood groups (1.6%). This study may enable us to contact individuals belonging to a particular blood group at times of medical emergencies when blood transfusion is required. Knowledge of blood group distribution is important for clinical studies, for reliable geographical information, blood bank management and for forensic studies in the population. Such a study would create awareness about self-blood grouping and also enable one to prepare a database of the available blood groups which can be utilized during medical emergencies for safe blood transfusion.

INTRODUCTION: Blood groups depend on antigens present on the surface of red blood cells. Scientists have discovered at least 30 common antigens and hundreds of rare antigens causing antigen-antibody reaction in human red blood cells. These antigens are genetically determined and are developed in fetal life and remain unchanged till death. Many blood group systems are identified but ABO and Rh blood groups are more antigenic.
 MATERIAL AND METHODS: In the present study, we observed ABO and Rh blood groups of 3057 students who studied in Universal College of Medical Sciences, Bhairahawa, Nepal from 1998 AD to 2019 AD, using open slide test method. The data was analyzed using SPSS version 20.
 RESULTS: Results of the present study indicated that the most common blood group was O (36.8%) followed by blood group B (31.1%) and blood group A (24.9%) and least common blood group was AB (7.2%) i.e. O>B>A>AB. The same sequence of ABO blood grouping was seen in both male and female. Rh positive blood group was found in 95.4% and Rh negative blood group was found in 4.6% of population.
 CONCLUSION: Knowledge of distribution of blood group is very important for medical students as they can serve as immediate blood donor in emergency conditions. This study can provide insight to advanced studies in future which can relate blood groups with medical conditions.