Accuracy and Performance Evaluation of Triplet Repeat Primed PCR as an Alternative to Conventional Diagnostic Methods for Fragile X Syndrome

Background@#Conventional diagnosis of fragile X syndrome (FXS) is based on a combination of fragment analysis (FA) and Southern blotting (SB); however, this diagnostic approach is time- and labor-intensive and has pitfalls such as the possibility of missing large number alleles. Triplet repeat primed PCR (TP-PCR) is a current alternative used to overcome these limitations. We evaluated the diagnostic usefulness of TP-PCR compared with the conventional diagnostic protocol consisting of FA and/or SB in terms of allele categorization, repeat number correlation, and zygosity concordance in female genetic carriers. @*Methods@#From November 2013 to March 2018, 458 patients (326 males, 132 females) were simultaneously examined using FA and/or SB and TP-PCR by detecting CGG repeat numbers in FMR gene and diagnosed as per American College of Medical Genetics guidelines. @*Results@#The TP-PCR results showed high concordance with the FA and/or SB results for all three aspects (allele categorization, repeat number correlation, and zygosity concordance in female genetic carriers). TP-PCR detected CGG expansions ≥ 200 in all full mutation (FM) allele cases in male patients, as well as both the normal allele (NL) and FM allele in female carriers. In premutation (PM) allele carriers, the TP-PCR results were consistent with the FA and/or SB results. In terms of zygosity concordance in female genetic carriers, 12 NL cases detected by TP-PCR showed a merged peak consisting of two close heterozygous peaks; however, this issue was resolved using a 10-fold dilution. @*Conclusions@#TP-PCR may serve as a reliable alternative method for FXS diagnosis.

Comparison of IRIS Iq200, UF-1000i, and Cobas u701 Module Automated Urine Sediment Analyzers

Background@#We sought to compare the performance of three commercially available automated urine sediment analyzers that represent the current urine sediment analysis technology. @*Methods@#A total of 232 patient samples were analyzed using manual microscopy and three automated analyzers: IRIS Iq200 (Beckman Coulter, USA), UF-1000i (Sysmex, Japan), and Cobas u701 (Roche, Switzerland). We analyzed precision, linearity, carry-over, concordance rate, and agreement between the three analyzers and manual microscopy. @*Results@#The repeatability and within-laboratory precision showed results similar to those of previous studies. All analyzers showed excellent linearity. The carry-over rates were within 1%. The correlation coefficient (r) between the three analyzers and manual microscopy was good. Regarding red blood cell (RBC), the UF-1000i showed a better concordance rate (90.52%) with manual microscopy than the other two analyzers and the agreement was substantial for UF-1000i (κ=0.63) and IRIS Iq200 (κ=0.61). Regarding white blood cell (WBC), Cobas u701 showed the best concordance rate (96.55%) and the agreement was moderate for IRIS Iq200 (κ=0.57) and Cobas u701 (κ=0.56), and fair for UF-1000i (κ=0.47). Regarding epithelial cell (EPI), IRIS Iq200 showed the highest concordance rate (99.2%) and the agreement was moderate for IRIS Iq200 (κ=0.59) and Cobas u701 (κ=0.54), and fair for UF-1000i (κ=0.40). @*Conclusions@#IRIS Iq200 offered the best agreement with manual microscopy for WBC and EPI count, while UF-1000i showed a better agreement for RBC count. The agreement is insufficient for fully replacing the manual microscopy.

Comparison of IRIS Iq200, UF-1000i, and Cobas u701 Module Automated Urine Sediment Analyzers

Background@#We sought to compare the performance of three commercially available automated urine sediment analyzers that represent the current urine sediment analysis technology. @*Methods@#A total of 232 patient samples were analyzed using manual microscopy and three automated analyzers: IRIS Iq200 (Beckman Coulter, USA), UF-1000i (Sysmex, Japan), and Cobas u701 (Roche, Switzerland). We analyzed precision, linearity, carry-over, concordance rate, and agreement between the three analyzers and manual microscopy. @*Results@#The repeatability and within-laboratory precision showed results similar to those of previous studies. All analyzers showed excellent linearity. The carry-over rates were within 1%. The correlation coefficient (r) between the three analyzers and manual microscopy was good. Regarding red blood cell (RBC), the UF-1000i showed a better concordance rate (90.52%) with manual microscopy than the other two analyzers and the agreement was substantial for UF-1000i (κ=0.63) and IRIS Iq200 (κ=0.61). Regarding white blood cell (WBC), Cobas u701 showed the best concordance rate (96.55%) and the agreement was moderate for IRIS Iq200 (κ=0.57) and Cobas u701 (κ=0.56), and fair for UF-1000i (κ=0.47). Regarding epithelial cell (EPI), IRIS Iq200 showed the highest concordance rate (99.2%) and the agreement was moderate for IRIS Iq200 (κ=0.59) and Cobas u701 (κ=0.54), and fair for UF-1000i (κ=0.40). @*Conclusions@#IRIS Iq200 offered the best agreement with manual microscopy for WBC and EPI count, while UF-1000i showed a better agreement for RBC count. The agreement is insufficient for fully replacing the manual microscopy.

Pitfalls of ABO Genotyping Based on Targeted Single Nucleotide Variant Analysis Due to a Nondeletional O Allele Lacking c.261delG: First Report of ABO*O.09.01 in Korea

No abstract available.

Changes in Antimicrobial Susceptibility Pattern of Blood Isolates at a University Hospital in the Kyungnam area during 2005–2014 / 고신대학교의과대학학술지

OBJECTIVES: Blood culture is a one of the most important procedure for diagnosis and treatment of infectious disease, but distribution of pathogenic species and the antimicrobial susceptibility can be vary from pathogen, individual trait, regional or environmental features. In this study, we investigated the changes in frequency of occurrence and antimicrobial susceptibility pattern of blood isolates from 2005 to 2014. METHODS: Data of blood isolates from Kosin Gospel Hospital during 2005 to 2014 were analyzed retrospectively. Blood isolates were cultured for 5 days using BACTEC Plus Aerobic/F and BACTEC lytic/10 Anaerobic/F. Identification and antimicrobial susceptibility test was performed using VITEK 1 system, VITEK 2 XL, PHOENIX 100 and conventional method. RESULTS: 9,847 isolates were identified during 10 years. Among the isolates aerobic or falcutative anaerobic bacteria were isolated in 99.5% specimens, anaerobic were 0.1%, and fugi were 0.4%. Most commonly isolated bacteria were coagulase-negative Staphylococcus (CoNS) followed by Escherichia coli, Staphylococcus aureus and Klebsiella pneumoniae. Candida parapsilosis were most frequently isolated among fungi. The proportion of S. aureus, A. baumannii and E. faecium were increased, while Pseudomonas aeruginosa and Streptococcus pneumoniae decreased over decennium. Imipenem resistant K. pneumoniae were identified. Vancomycin resistant E. faecium and imipenem resistant A. baumannii were increased (7.1% in 2005 to 12.3% in 2014, 0% in 2005 to 55.6% in 2014, respectively). CONCLUSIONS: Over the last 10 year, CoNS were the most frequently isolated pathogen. Imipenem resistant K. pneumoniae was emerged. Vancomycin resistant E. faecium and imipenem resistant A. baumannii increased during this period.

A Case of Therapy-Related Acute Leukemia With Mixed Phenotype With BCR-ABL1 After Treatment of Diffuse Large B-Cell Lymphoma

No abstract available.

Comparison of Three Commercially Available Assays for Measurement of Vitamin D

BACKGROUND: We evaluated three commercially available vitamin D assays to evaluate and compare the correlation and accuracy among them. METHODS: Vitamin D was measured in 71 patient samples using the Architect 25-OH vitamin D assay (Abbott), the ADVIA Centaur vitamin D total assay (Siemens), and the LIAISON 25 OH vitamin D total assay (Diasorin). The evaluation made use of both patient samples and standard reference material, SRM 972. To analyze correlations and differences, Pearson's correlation coefficients and paired sample t-tests were performed. RESULTS: Correlations among the three evaluated assays showed strong positive linear relationships (correlations among Siemens and DiaSorin, DiaSorin and Abbott, Abbott and Siemens: r=0.935, r=0.927, r=0.909, respectively). Mean (SD) vitamin D values on Siemens, Abbott, and DiaSorin assays in the 71 patient samples were 23.09 (10.41), 16.75 (11.26), and 16.76 (9.32), respectively. Results for the Siemens assay were significantly different from the other two methods (P<0.001). Target values for SRM 972 level 1, 2, 3, and 4 were 23.9, 14.0, 44.9, and 35.4, respectively. The Abbott, Siemens, and Diasorin assay values were closest to the target values in level 1, levels 2 and 3, and level 4, respectively. CONCLUSIONS: Correlations among vitamin D assays were good; however, the mean values of the Siemens assay were significantly higher than those of DiaSorin or Abbott. We found significant differences in vitamin D levels and discrepancies between patient samples and SRM 972 samples, which should be considered during use in a clinical setting.

Falsely Elevated Tacrolimus Concentrations Using Chemiluminescence Microparticle Immunoassay in Kidney Transplant Patient / 대한이식학회지

Tacrolimus is one of the effective immunosuppressive drugs used after an organ transplant procedure. However, due to its narrow therapeutic range, its usefulness in preventing transplant rejection and minimizing nephrotoxicity is dependent on the monitoring of whole blood trough levels of tacrolimus. A 49-year-old kidney transplant recipient presenting with cough and general weakness was admitted to the hospital. Due to the patient's deeply compromised clinical condition, an immunosuppressive therapy was discontinued. Tacrolimus concentrations in the patient's whole blood samples were measured, using an automated chemiluminescent microparticle immunoassay (CMIA) instrument. Interference was suspected because tacrolimus concentrations after the discontinuation of tacrolimus dose were 20.9 and 18.2 ng/mL at day 2 and 3, respectively. Tacrolimus concentrations were 11.1 and 12.6 ng/mL, respectively, when re-tested using an antibody-conjugated magnetic immunoassay (ACMIA). We evaluated the relationship between the CMIA and ACMIA results, and calculated the expected values from the regression equation. Residuals were –8.4 and –4 ng/mL, respectively. There have been several cases with false detection of elevated tacrolimus concentrations using ACMIA; however, such falsely detected elevations using CMIA have rarely been reported. When unexpectedly high concentrations of tacrolimus are detected by CMIA in transplant patients, an immediate re-test using another technique might be necessary to rule out falsely elevated results.