Molecular Typing for Kidd Blood Group in Taiwan.

Blood ◽  
2005 ◽  
Vol 106 (11) ◽  
pp. 4174-4174
Author(s):  
Cheng-Hwai Tzeng ◽  
Jau-Yi Lyou ◽  
Pei-Shan Chen ◽  
Hsueng-Mei Liu ◽  
Ying-Ju Chen ◽  
...  

Abstract Background:The Kidd (JK) blood group system is clinically important intransfusion medicine. Alloantibodies to antigens in this system may be produced following blood transfusion or during pregnancy and can result in serious hemolytic transfusion reactions and hemolytic disease of the newborn (HDN). The two major codominant alleles of the JK gene, Jka and Jkb, have a similar frequency in Caucasian populations (0.51 and 0.49, respectively) and define 3 common phenotypes Jk(a+b+), Jk(a+b-), Jk(a-b+), while frequency of Jk(a-b-) or Jknull phenotype is exceedingly rare. Many individuals of Polynesian extraction have been identified as Jknull. Its frequency and molecular characterization for Chinese people in Taiwan, however, has not yet been clarified. Study design and methods: By using the lately developed single-tube allele-specific primer/multiplex PCR technique, a total of 320 random whole blood samples were analyzed. In addition, three samples already serologically proven as Jknull phenotype in our Lab were also investigated. Results: None of the 320 random whole blood samples was serologically typed as Jk(a-b-), while the frequencies of three other phenotypes were 47.5% for Jk(a+b+), 23.1% for Jk(a+b-), and 29.4% for Jk(a-b+). Interestingly, two Jka/Jk and four Jkb/Jk were identified, resulting in a gene frequency of Jka - 46.72%, Jkb - 52.34% and silent Jk - 0.94%. As for the three samples of Jknull phenotype all belonged to the so-called Polynesian type, i.e.3′-acceptor splice site G->A mutation of intron 5 that resulted in the skipping of exon 6 (called mutation JKΔ6). Conclusion: The present study confirms that the single-tube allele-specific primer/multiplex PCR technique has enabled genomic typing of the Kidd blood group easier and reliable. The frequency and molecular characterization of JK phenotypes for Chinese people in Taiwan have been clearly characterized.

Blood ◽  
2010 ◽  
Vol 116 (21) ◽  
pp. 3348-3348
Author(s):  
Cassandra Josephson ◽  
John Roback ◽  
Robert Myers ◽  
Lisa Hallock ◽  
Irene DeMezzo ◽  
...  

Abstract Abstract 3348 Background: Technologies have recently been developed for rapid determination of extended human erythrocyte antigen (xHEA) phenotypes. For example, a semi-automated method using allele-specific oligonucleotides targeted against 32 clinically significant minor RBC antigens has been used to determine donor xHEA phenotypes from whole blood samples. This approach is currently used by blood collection centers and medical centers with blood collection facilities (both sites have access to linked donor whole blood samples). Broader access to xHEA information closer to the point-of-care (e.g. Transfusion Services at a Medical Center without a blood collection facility) may provide an opportunity to enhance patient care by more quickly and broadly providing units with xHEA phenotypes (Klapper et al., 2010.) However, transfusion services would need to use integrally attached segments for testing, and with leukoreduced (LR) RBC units these segments have very low numbers of white blood cells (WBC) (and therefore DNA), potentially limiting analysis. This study was performed to determine whether a HEA-elongation mediated multiplex assay in solution (HEA-eMAP-S) (Xin et al., 2010) could accurately genotype segments from LR-RBC units for 32 clinically significant minor RBC antigens. Methods: Segments from pre-storage LR-RBC units (American Red Cross), < 14 days old, were obtained from a large tertiary care Children's Hospital in the Southeastern US and residual WBC were quantified by flow cytometry. DNA was extracted using an extraction method developed at BioArray SolutionS (BAS) using commercial reagents (Qiagen, Inc., Valencia, CA), and then amplified with the Universal Beadchip™ package (HEA LR-eMAP-S Beadchip™ Kits) which contains allele specific oligonucleotides directed to 32 clinically significant blood group antigens (c, C, e, E, V, VS, K, k, Kpa, Kpa, Jsa, Jsb, Jka, Jkb, Fya, Fyb, M, N, S, s, Lua, Lub, Dia, Dib, Coa, Cob, Doa, Dob, Joa, Hy, Yta, Ytb mutation for hemoglobin S). DNA analysis results were correlated with RBC storage solution, WBC filter type, and serologic minor RBC antigen phenotypes of the units. Results: 102 LR-RBC units from whole blood donations were studied, 74 /102 (73 %) stored in AS-1 and 28 /103 (27 %) in CPDA-1 solution. All AS-1 units were pre-storage LR with Fenwal Sepacell Flex Excel Filters and all CPDA-1 units were pre-storage LR with Whole Blood Fenwal Filters (Fenwal Inc. Lake Zurich, IL). All units demonstrated < 5 × 106 WBC/unit with 47 % having < 4 × 104 WBC/unit, which is at or below the limit of flow cytometric detection. Complete genotyping data was obtained from all samples. Ten samples showing initial indeterminate results on Diego and one for N antigens produced complete results after repeat testing. Fifty-four percent of units were serologically phenotyped for 1–8 antigens by the blood collection center; there was 100% correlation between predicted phenotype from DNA analysis and serology for these units. Conclusions: The HEA LR-eMAP-S DNA analysis can be applied to optimally pre-storage LR-RBC units yielding > 99 % accuracy for all minor red blood cell antigens tested. The ability to perform this type of testing in a hospital transfusion service opens up new possibilities for transfusion services to select from their existing inventory and more efficiently allocate units to recipients with specific phenotypic requirements for RBC units. Disclosures: Josephson: Immucor: Speakers Bureau. DeMezzo:Immucor: Employment. Tanzi:Immucor: Employment. Enriquez:Immucor: Employment. Lin:Immucor: Employment. Hashmi:Immucor: Employment.


1999 ◽  
Vol 37 (6) ◽  
pp. 2020-2023 ◽  
Author(s):  
S. A. Gilbert ◽  
K. M. Burton ◽  
S. E. Prins ◽  
D. Deregt

A nested multiplex PCR was developed for genotyping of bovine viral diarrhea viruses (BVDVs). The assay could detect as little as 3 50% tissue culture infective doses of BVDV per ml and typed 42 out of 42 cell culture isolates. BVDV was also successfully typed, with or without RNA extraction, from all 27 whole-blood samples examined from 22 carriers or probable carriers and 5 experimentally infected cattle.


2013 ◽  
Vol 319 ◽  
pp. 169-173
Author(s):  
Chun Fang Wang ◽  
Yun Hang Gao ◽  
Xiu Yun Jiang ◽  
Jian Wang ◽  
Gong Mei Li ◽  
...  

For molecular biological detection of Mycobacterium tuberculosis, PCR methods with primers targeting different regions specific for mycobacterium tuberculosis complex are used worldwide. In this study,256 whole blood samples were detected using multiplex PCR.2 of all samples were found to be PCR positive. Homogenous comparison of Mycobacterium bovis specific genes with Mycobacterium bovis AF2122/97 all reached up to 99%.


2010 ◽  
Vol 41 (02) ◽  
Author(s):  
N Shazi ◽  
A Böss ◽  
HJ Merkel ◽  
F Scharbert ◽  
D Hannak ◽  
...  

Separations ◽  
2021 ◽  
Vol 8 (6) ◽  
pp. 78
Author(s):  
Sevasti Karampela ◽  
Jessica Smith ◽  
Irene Panderi

An ever-increasing need exists within the forensic laboratories to develop analytical processes for the qualitative and quantitative determination of a broad spectrum of new psychoactive substances. Phenylethylamine derivatives are among the major classes of psychoactive substances available on the global market and include both amphetamine analogues and synthetic cathinones. In this work, an ultra-high-performance liquid chromatography-positive ion electrospray ionization tandem mass spectrometric method (UHPLC-ESI-MS/MS) has been developed and fully validated for the determination of 19 psychoactive substances, including nine amphetamine-type stimulants and 10 synthetic cathinone derivatives, in premortem and postmortem whole blood. The assay was based on the use of 1 mL premortem or postmortem whole blood, following solid phase extraction prior to the analysis. The separation was achieved on a Poroshell 120 EC-C18 analytical column with a gradient mobile phase of 0.1% formic acid in acetonitrile and 0.1% formic acid in water in 9 min. The dynamic multiple reaction monitoring used in this work allowed for limit of detection (LOD) and lower limit of quantitation (LOQ) values of 0.5 and 2 ng mL−1, respectively, for all analytes both in premortem and postmortem whole blood samples. A quadratic calibration model was used for the 12 quantitative analytes over the concentration range of 20–2000 ng mL−1, and the method was shown to be precise and accurate both in premortem and postmortem whole blood. The method was applied to the analysis of real cases and proved to be a valuable tool in forensic and clinical toxicology.


2021 ◽  
Vol 2 (1) ◽  
pp. 100311
Author(s):  
Daniella C. Terenzi ◽  
Ehab Bakbak ◽  
Justin Z. Trac ◽  
Mohammad Al-Omran ◽  
Adrian Quan ◽  
...  

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