Sequence variants in the proximal promoter and +5.8-kb site of ABO in Koreans with weak B phenotypes.

BACKGROUND AND OBJECTIVES: Several studies on Chinese and Japanese populations have revealed that a substantial proportion of weak B subgroups are caused by variants in the major regulatory regions of ABO, the proximal promoter, CCAAT-binding factor/NF-Y binding site and +5.8-kb site. We performed molecular analyses of these regions in Koreans with weak B phenotypes. MATERIALS AND METHODS: This study included 16 samples with weak B phenotypes (4 B3 , 1 Bw , 5 A1 B3 and 6 A1 Bw ) harbouring no subgroup-causing variants in ABO exons 6 and 7. These samples were subjected to sequencing analysis of exons 1-5 and the major regulatory regions of ABO. RESULTS: Of the 16 samples, 14 were found to carry a sequence variant either in the proximal promoter (g.4991_5008del [n = 3]) or the +5.8-kb site (g.10893G>A [n = 4] and g.10925C>T [n = 7]). The remaining two samples were found to contain no subgroup-causing variants. CONCLUSION: Our study demonstrates that sequence variants in the proximal promoter and +5.8-kb site account for a substantial proportion of weak B subgroups in Koreans, suggesting that molecular analysis of these regions is essential for the accurate determination of ABO genotypes in Koreans with weak B phenotypes.

Analysis of ABO grouping discrepancies among patients from a tertiary hospital in Korea.

BACKGROUND: Accurate ABO typing is essential for preventing ABO incompatibility reactions. However, the causes of ABO grouping discrepancy has not been sufficiently studied, and it may vary among different ethnic populations. Thus, the aim of this retrospective study was to investigate the causes of ABO discrepancy in the East Asian population. MATERIALS AND METHODS: A retrospective observational study on ABO typing discrepancy among patients in a tertiary hospital was carried out using the electronic medical record database of Samsung Medical Center (Seoul, Korea) between July 2016 and May 2019. RESULTS: ABO grouping was performed on 551,959 blood samples during the study period; 1468 events of serologic ABO discrepancy were determined from 1334 (0.24 %) samples. A total of 134 samples (0.02 %) presented multiple causes of ABO discrepancy. Weak/missing serum reactivity (594, 40.5 %) was the most frequent reason for ABO discrepancy, followed by extra serum reactivity (370, 25.2 %), weak/missing red cell reactivity (267, 18.2 %), mixed-field red cell reactivity (176, 12.0 %), and extra red cell reactivity (61, 4.2 %). In the category of weak/missing red cell reactivity, ABO subgroup was the most common reason, and using ABO genotyping, 26.2 % of the cases genotyped were found to be related to the cis-AB allele. CONCLUSIONS: Our results suggest that the incidence and cause of ABO typing discrepancies vary among institutes and ethnic groups. Our data helps to better understand and facilitate the resolution of ABO typing discrepancies in patients.

Evaluation of the ASTA MicroIDSys matrix-assisted laser desorption ionization time-of-flight mass spectrometry system for identification of mycobacteria directly from positive MGIT liquid cultures.

OBJECTIVES: We evaluated the performance of the MicroIDSys Elite system, a newly developed matrix-assisted laser desorption ionization time-of-flight (MALDI-TOF) mass spectrometry system for identification of mycobacteria directly from positive MGIT liquid cultures. METHODS: Analytical specificity was evaluated with 63 reference strains grown in mycobacteria growth indicator tube media. Prospective performance evaluation was conducted with primary liquid cultures of sputum samples for identification of mycobacteria, and results were compared to multigenerational sequencing as the reference method. Liquid media subcultures were also analyzed. RESULTS: The accuracy for the 63 reference strains was 98.4% (62/63). A total of 167 paired mycobacterial primary cultures and subcultures in liquid media, comprised of seven Mycobacterium tuberculosis isolates, 109 slowly growing nontuberculous mycobacterial isolates, and 51 rapidly growing nontuberculous mycobacterial isolates, was identified by the MicroIDSys Elite system. Using primary liquid cultures, the MicroIDSys Elite system correctly identified 143 (85.6%) isolates; 21 (12.6%) resulted in "no identification"; and three (1.8%) isolates were misidentified. Using liquid media subcultures with this system, 159 (95.2%) isolates were correctly identified; seven (4.2%) resulted in "no identification"; and one (0.6%) isolate was misidentified. CONCLUSION: The MicroIDSys Elite system is a useful routine diagnostic tool for identification of mycobacterial species from liquid culture.

Molecular basis of serological weak D phenotypes and RhD typing discrepancies identified in the Korean population.

BACKGROUND: The molecular basis of RhD blood groups differs with race/ethnicity. This study aimed to investigate the molecular basis of serological weak D phenotypes and RhD typing discrepancies in the Korean population. MATERIALS AND METHODS: The RhD status of 188,852 Korean patients was initially determined using the automated microplate method and manual tile method. In case of no agglutination, weak D testing was further performed using the tube and gel methods. Serologically D-negative samples with C+ and/or E+ were tested using polymerase chain reaction-sequence specific primers for four RHD targets and/or exon 9 sequencing. Samples showing a serological weak D phenotype or an RhD typing discrepancy were subjected to full RHD gene sequencing. RESULTS: Of the 32 samples showing a serological weak D phenotype and 191 samples showing a serologically D-negative phenotype with C+ and/or E+, 23 and 50 were genotyped, respectively. Among the weak D samples, the most common alleles were RHD*15 (n=6), RHD*13.01 (n=4), and RHD*01W.25 (n=4), and no variant was found in two samples. RHD*01EL.01 (n=26) accounted for more than half of the D-negative samples. Of the seven samples that were typed as D-positive using the automated microplate method but showed weak reactivity using the tile method, four were genotyped, and the results were as follows: RHD*01W.33 (n=2), RHD*01W.43 (n=1), and no variant found (n=1). DISCUSSION: In our cohort, various D variant alleles including RHD*15 were identified; however, RHD*01W.1, RHD*01W.2, RHD*01W.3, RHD*09.03.01, and RHD*09.04, accounting for more than 95% of Caucasians with a serological weak D phenotype, were not found. Our study reaffirms that the distribution of D variant alleles differs between East Asians and Caucasians. Our findings also indicate that some D variants including RHD*01W.33 and RHD*01W.43 are at risk of being mistyped as D-positive by a highly sensitive RhD typing method such as an automated microplate method.

Performance evaluation of Automated Fluorescent Immunoassay System ROTA and NORO for detection of rotavirus and norovirus: A comparative study of assay performance with RIDASCREEN® Rotavirus and Norovirus.

BACKGROUND: The Automated Fluorescent Immunoassay System ROTA (AFIAS-Rota) and NORO (AFIAS-Noro) assays (Boditech Med Inc.) are newly developed diagnostic tests for rotavirus and norovirus infections. METHODS: Performance of AFIAS-Rota/Noro assays was evaluated in comparison with RIDASCREEN® Rotavirus and Norovirus ELISA kits (R-Biopharm) using clinical stool samples submitted from November 2018 to January 2019. Multiplex real-time reverse transcription-polymerase chain reaction was used as reference method. RESULTS: A total of 256 clinical specimens were analyzed. AFIAS-Rota and RIDASCREEN Rotavirus had almost perfect agreement (Kappa value = 0.95), and substantial agreement was observed between AFIAS-Noro and RIDASCREEN Norovirus (Kappa value = 0.80). For detection of rotavirus, AFIAS and RIDASCREEN assays showed satisfactory diagnostic sensitivity (100% and 97.8%, respectively) and specificity (99.5% and 99.1%). For detection of norovirus, the RIDASCREEN assay showed significantly higher sensitivity than the AFIAS-Noro (86.0% and 66.0%, respectively; P = .002). Analytic specificity of AFIAS-Rota/Noro assays showed no cross-reactivity against any other bacteria (14 strains) or viruses (2 strains). Hands-on time (6 minutes) and turnaround time (26 minutes) required to perform AFIAS assays were much shorter than those required for RIDASCREEN assays (20 and 150 minutes, respectively). CONCLUSION: The AFIAS-Rota/Noro assays showed overall excellent agreement with the RIDASCREEN assays. Although the AFIAS-Noro assay exhibited lower sensitivity than the RIDASCREEN Norovirus assay for detection of norovirus, the AFIAS-Rota/Noro assays could be useful as a rapid initial screening test in clinical laboratories due to its convenience and rapid turnaround time.

Evaluation of the BioFire FilmArray Pneumonia Panel for rapid detection of respiratory bacterial pathogens and antibiotic resistance genes in sputum and endotracheal aspirate specimens.

OBJECTIVES: The performance of the investigational-use-only version of the BioFire FilmArray Pneumonia Panel (FA-Pneumo), a high-order nested multiplex PCR, was evaluated for the detection of typical respiratory bacterial pathogens and antibiotic resistance genes in sputa and endotracheal aspirate (ETA) specimens. METHODS: Thirty-one sputa and 69 ETA specimens were analyzed. The diagnostic performance of FA-Pneumo was assessed using routine microbiological methods as the reference standard. RESULTS: Overall sensitivity and specificity for organism detection using FA-Pneumo were 98.5% and 76.5%, respectively. The sensitivity for each pathogen was 100%, except for Klebsiella aerogenes, and the range of specificity was 83.3-99.0%. FA-Pneumo detected antimicrobial resistance genes in 17 out of 18 specimens (94.4%) that were resistant by antimicrobial susceptibility testing. FA-Pneumo additionally detected 25 resistance genes in 22 specimens, and sequencing for the presence of resistance genes confirmed the majority of these results (20/25, 80%). Semi-quantitative analysis of bacterial nucleic acid amounts by FA-Pneumo revealed that 88.2% of the identified bacteria (67/76) with ≥106 copies/ml also gave culture-positive results with significant amounts of bacteria. CONCLUSIONS: FA-Pneumo is a rapid test with high sensitivity for the detection of bacteria and antimicrobial resistance genes in sputum and ETA specimens and could aid in determining antibiotic therapy.

The first case of congenital blood chimerism in two of the triplets in Korea.

BACKGROUND: Chimeras are composed of two or more different populations that originated from different zygotes. Blood chimerism arising from twins have been reported in the literature. Herein, we report the first blood group chimerism in triplets. METHODS: ABO blood grouping was carried out by manual tile methods (Merck Millipore, UK) and micro-column agglutination method (Bio-Rad, Cressier sur Morat, Switzerland). Flow cytometric analysis was performed with Anti-A-PE conjugated monoclonal antibodies (BD Biosciences, San Jose, CA, USA) and FACS Canto II (BD Biosciences). Molecular analysis was performed with allele-specific polymerase chain reaction (AS-PCR) and direct sequencing of the exons 6 and 7. RESULTS: Mixed-field agglutination and weak agglutination against anti-A were revealed by ABO blood grouping. Flow cytometric analysis revealed the presence of both A cells and O cells. AS-PCR and sequencing showed two neonates with chimerism, with each neonate`s genotype being A102/O01/O02. CONCLUSION: This is the first recorded case of blood chimera from a triplet in Korea. We recommend full investigation of blood group chimerism in neonates with ABO discrepancy, as blood chimerism is subject to certain caution in the clinical environment.