The evaluation of histo-blood group ABO typing by flow cytometric and PCR-amplification of specific alleles analyses and their application in clinical laboratories

Aki, Kensaku; Izumi, Azusa; Hosoi, Eiji

doi:10.2152/jmi.59.143

Cited by 6 publications

(4 citation statements)

References 22 publications

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“…15,16 However, these studies were usually performed in fixed cells and by indirect fluoroimmunoassays, using secondary antibodies, which can make the experiments more laborious and less specific. Our methodology with CdTe QDs enabled us to quantitatively evaluate antigens on RBC membranes in A 1 , A 2 , B, A 1 B, O, and in some A weak groups both by profile and by labeling efficiency.…”

Section: Cabral Filho Et Almentioning

confidence: 99%

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Blood group antigen studies using CdTe quantum dots and flow cytometry

Filho¹,

Pereira²,

Fernandes³

et al. 2015

IJN

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New methods of analysis involving semiconductor nanocrystals (quantum dots [QDs]) as fluorescent probes have been highlighted in life science. QDs present some advantages when compared to organic dyes, such as size-tunable emission spectra, broad absorption bands, and principally exceptional resistance to photobleaching. Methods applying QDs can be simple, not laborious, and can present high sensibility, allowing biomolecule identification and quantification with high specificity. In this context, the aim of this work was to apply dual-color CdTe QDs to quantify red blood cell (RBC) antigen expression on cell surface by flow cytometric analysis. QDs were conjugated to anti-A or anti-B monoclonal antibodies, as well as to the anti-H ( Ulex europaeus I) lectin, to investigate RBCs of A 1 , B, A 1 B, O, A 2 , and A weak donors. Bioconjugates were capable of distinguishing the different expressions of RBC antigens, both by labeling efficiency and by flow cytometry histogram profile. Furthermore, results showed that RBCs from A weak donors present fewer amounts of A antigens and higher amounts of H, when compared to A 1 RBCs. In the A group, the amount of A antigens decreased as A 1 > A 3 > A X = A el , while H antigens were A X = A el > A 1 . Bioconjugates presented stability and remained active for at least 6 months. In conclusion, this methodology with high sensibility and specificity can be applied to study a variety of RBC antigens, and, as a quantitative tool, can help in achieving a better comprehension of the antigen expression patterns on RBC membranes.

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Section: Cabral Filho Et Almentioning

confidence: 99%

“…Flow cytometric analysis of RBC samples aBO blood group incubated with QDs-anti-a and QDs-anti-B Table 2 15 and by Hult and Olsson. 16 The percentage of labeled RBCs and the labeling profile were similar to ours in both studies, where the authors analyzed ABO antigens stained by a secondary antibody conjugated to organic dyes using an indirect immunoassay.…”

mentioning

confidence: 99%

Blood group antigen studies using CdTe quantum dots and flow cytometry

Filho¹,

Pereira²,

Fernandes³

et al. 2015

IJN

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show abstract

“…Patient samples were amplified by exons 6 and 7 of the ABO gene and then sequenced using an ABO exons 6 and 7 sequence kit (Jiangsu ZojiWat Biomedical Co., Ltd. China). The sequencing primers can be observed in the literature [3] . The measured gene sequence was compared with the ABO * A101 structure on file (GenBank serial number: AJ536122) using Seq Scape 3 software [5] .…”

Section: Genetic Sequencing Testmentioning

confidence: 99%

“…Often, the identification of rare ABO subtypes lacks accuracy and timeliness due to limitations inherent in traditional serological methods, which may easily cause serious harm to health [2] . The application of molecular biological methods for ABO blood groups, combining the method of serology and gene detection, is beneficial to the accurate identification of ambiguous ABO blood groups and guarantees blood transfusion safety [3] . This paper summarizes the process of using genetic analytical methods to determine the ABO blood group of 21 patients with inconsistent forward and reverse ABO group typing from the samples collected in the First Affiliated Hospital of Nanjing Medical University from December 2018 to December 2019.…”

Section: Introductionmentioning

confidence: 99%