Prevalence and Clinical Relevance of Exon 2 Deletion of COMMD1 in Bedlington Terriers in Korea.

BACKGROUND: Deletion of exon 2 of copper metabolism domain containing 1 (COMMD1) results in copper toxicosis in Bedlington terriers (CT-BT). OBJECTIVES: This study was conducted to identify the prevalence and clinical relevance of the COMMD1 mutation in Bedlington terriers in Korea. ANIMALS: A total of 105 purebred Bedlington terriers (50 males, 55 females) from the kennels and pet dog clubs in Korea were examined during the period 2008-2013. METHODS: A multiplex PCR was carried out to detect exon 2 deletion of COMMD1. Clinical analysis was performed on each genetic group, and clinical status of the dogs was followed up to estimate survival probability. RESULTS: Of the 105 samples, 52 (49%) were wild-type homozygote, 47 (45%) were heterozygote, and 6 (6%) were mutant-type homozygote. Plasma alanine aminotransferase (ALT) activity was increased in the mutant-type homozygous group >2 years of age (P < .0001). The survival probability of 6 mutant-type homozygotes surviving 2.5 years was 0.67, and 4 years was 0.5. CONCLUSIONS AND CLINICAL IMPORTANCE: Results show the prevalence and clinical relevance of exon 2 deletion of COMMD1 and could help establish a structured selective breeding program to prevent CT-BT in Korea.

Comparison of white blood cell counts by WNR, WDF, and WPC channels in Sysmex XN hematology analyzer.

INTRODUCTION: The Sysmex XN modular system (Sysmex, Kobe, Japan) uses a novel technology for white blood cell (WBC) count and differential, using separate channels: white cell nucleated (WNR), WBC differential (WDF), and white progenitor cell (WPC) channels. We questioned how concordant WBC counts would be between them. METHODS: In a total of 6327 consecutive specimens, WBC counts were compared between WNR and WDF channels. They were also compared in three groups of WBC counts and two groups of chemotherapy status. In 508 specimens from the 4361 specimens that were run on the XN-20 module, the WPC channel was used for reflex test. Data were compared using Pearson's correlation, absolute difference, and percent difference (%D). RESULTS: WBC counts between WNR and WDF channels showed very high correlations in total specimens (r = 0.9976) and in the groups of WBC counts and chemotherapy. As WBC count increased, absolute difference increased, while %D decreased (P < 0.0001, both). Percent difference was 1.55% in total specimens and showed the highest value in the severe leukopenia group (<1.0 × 10(9)/L, 6.18%). CONCLUSIONS: This is the first large-scale study on novel channel technology for WBC counts in the Sysmex XN. WBC counts by WNR, WDF, and WPC channels are highly correlated, and they are overall interchangeable and reliable.

Evaluation of ABX Pentra DX 120 and Sysmex XE-2100 in umbilical cord blood.

INTRODUCTION: Cord blood (CB) is an important source of hematopoietic stem cells and reflects the hematologic status of neonates. ABX Pentra DX 120 (Horiba Medical, Montpellier, France) and Sysmex XE-2100 (Sysmex, Kobe, Japan) were compared in 200 CB specimens. METHODS: Complete blood count parameters including white blood cell (WBC) differential counts were compared between the two analyzers. Double differential matrix (DDX) by ABX Pentra DX 120 and hematopoietic progenitor cell (HPC) by Sysmex XE-2100 were compared with CD34(+) cells by flow cytometry. RESULTS: Most of the parameters showed acceptable correlation between the two analyzers. Although WBC differential of both analyzers showed acceptable correlation with manual counts, mononuclear cells (MNC) by ABX Pentra DX 120 better correlated with manual count than MNC by Sysmex XE-2100. NRBC by Sysmex XE-2100 better correlated with manual count than NRBC by ABX Pentra DX 120. ABX Pentra DX 120 showed better flagging performances. DDX better correlated with CD34(+) cells than HPC. CONCLUSION: Although the results from both analyzers are mostly interchangeable and reliable in CB specimens, flagging performance of ABX Pentra DX 120 seems to be superior to that of Sysmex XE-2100. DDX by ABX Pentra DX 120 would be valuable to evaluate the quality of CB for further therapeutic utilization.

Establishment of reference interval for immature platelet fraction.

INTRODUCTION: Immature platelet fraction (IPF) is a parameter for reticulated platelets. A high percentage IPF (%-IPF) is indicative of consumptive or recovering thrombocytopenic disorders in contrast to a low %-IPF seen in aplastic states. Absolute IPF (A-IPF) specifically reflects the number of immature platelets in circulation. This study aimed to establish reliable reference intervals for %-IPF and A-IPF. METHODS: Except outliers, platelet counts and IPF were determined in 2152 healthy individuals (1252 men and 900 women) and 133 umbilical cord blood from healthy full-term neonates using XE-2100 hematology analyzer (Sysmex, Kobe, Japan). The reference intervals for %-IPF and A-IPF were defined using nonparametrical percentile methods according to the Clinical and Laboratory Standard Institute (CLSI) guideline. RESULTS: Platelets,%-IPF, and A-IPF all showed nonparametrical distributions. In total individuals, the reference intervals for %-IPF and A-IPF were 0.5-3.3% (0.5-3.1% in men; 0.5-3.4% in women) and 1.25-7.02 × 10(9) /L (1.30-6.80 × 10(9) /L in men; 1.21-7.15 × 10(9) /L in women), respectively. The reference intervals for %-IPF and A-IPF in umbilical cord blood were 0.7-3.8% and 1.93-9.7 × 10(9) /L, respectively. CONCLUSIONS: This study provides the reference interval for IPF, including %-IPF and A-IPF, according to the CLSI guideline. These results could be used as fundamental data for clinical use as well as future researches.

Fecal carriage of extended-spectrum ß-lactamase-producing Enterobacteriaceae in Korean community and hospital settings.

PURPOSE: The assessment and early recognition of risk factors for infections due to extended-spectrum ß-lactamase-producing Enterobacteriaceae (ESBL-E) are important for infection control and proper treatment. The aim of the present study was to investigate the prevalence of fecal carriage of ESBL-E in healthy individuals and hospitalized high-risk patients in Korea and to compare the characteristics of ESBL-E in these two groups. METHODS: A total of 384 samples from 290 healthy individuals and 94 high-risk patients were collected. The screening of ESBL-E was performed using a commercial chromogenic medium. Bacterial identification and antibiotic susceptibility testing were performed using the Vitek 2 system. RESULTS: The prevalence of ESBL-E carriage was 20.3 % in healthy individuals and 42.5 % in high-risk patients. Escherichia coli comprised a large majority (96.6 %) of the isolates from healthy individuals, but Klebsiella pneumoniae was more commonly detected (45.0 %) in high-risk patients than in healthy individuals. K. pneumoniae isolates exhibited significantly higher resistance to ceftazidime, ampicillin, and carbapenem, and E. coli exhibited higher resistance to cefotaxime. E. coli from high-risk patients exhibited significantly higher resistance to levofloxacin and cefepime than that from healthy individuals. CONCLUSIONS: We demonstrated the high prevalence of ESBL-E carriage in Korea and clarified the characteristics of ESBL-E carriage in healthy individuals and high-risk patients. The distribution and antibiotic susceptibility of colonizing ESBL-E were different between the group of healthy individuals and the high-risk patients. Active surveillance of ESBL-E carriage is suggested for infection control, and the use of chromogenic agar appears to be an efficient method.

Isothermal target and probe amplification for Mycobacterium tuberculosis identification from broth cultures.

SETTING: Various methods are used to identify Mycobacterium tuberculosis complex (MTC) from broth cultures. The isothermal target and probe amplification (iTPA) method has recently been introduced as a simple and cost-effective molecular assay. OBJECTIVE: To evaluate the diagnostic performance of the iTPA method. DESIGN: A total of 175 specimens from the Mycobacteria Growth Indicator Tube (MGIT) 960 broth culture system were evaluated. The immunochromatographic test (ICT) and real-time quantitative PCR (RQ-PCR) were compared with the iTPA method. RESULTS: MTC was identified in 71/131 MGIT-positive specimens, including 60 ICT-positive and 11 ICT- negative/PCR-positive specimens. The sensitivity and specificity of the ICT assay were respectively 84.5% (95%CI 74.0-92.0) and 100% (95%CI 94.0-100). These 71 specimens were all MTC-positive with the iTPA method also. Sixty non-tuberculous mycobacteria specimens and 44 MGIT-negative specimens were all MTC-negative with the iTPA method. CONCLUSION: Our data show that the diagnostic performance of the iTPA method is comparable to that of RQ-PCR. The iTPA method could be a reliable and cost-effective option for the identification of MTC from broth culture.

Detection of tuberculosis using artus M. tuberculosis PCR Kit and COBAS AMPLICOR Mycobacterium tuberculosis Test.

SETTING: Nucleic acid amplification tests can detect Mycobacterium tuberculosis complex rapidly and reliably. OBJECTIVE: To compare the diagnostic performance of the artus M. tuberculosis PCR Kit and COBAS AMPLICOR Mycobacterium tuberculosis Test. In the artus assay, an appropriate cycle threshold (Ct) value was determined for positivity. DESIGN: A total of 238 clinical respiratory specimens were analysed using both the artus and COBAS AMPLICOR assays. In 221 specimens, these results were further compared with culture results. RESULTS: The overall agreement between artus and COBAS AMPLICOR was 96.2% (229/238). Among the nine (3.8%) discrepant specimens, three (1.3%) were artus-positive and COBAS AMPLICOR-negative, while the other six (2.5%) were artus-negative and COBAS AMPLICOR-positive. Using culture as a standard, the sensitivity and specificity of the artus assay were 97.8% and 85.1%, and those of COBAS AMPLICOR assay were 100% and 86.2%, respectively. The difference was not statistically significant. In the artus assay, the minimum Ct value for the positivity determination was 38. CONCLUSION: The artus and COBAS AMPLICOR assays showed comparable diagnostic performance and can be confidently used for detection of M. tuberculosis complex. In the artus assay, a Ct value of 38 could be suggested as an appropriate cut-off value.

Optimization of laboratory workflow in clinical hematology laboratory with reduced manual slide review: comparison between Sysmex XE-2100 and ABX Pentra DX120.

INTRODUCTION: The validation of automated hematology analyzer results by manual slide review (MSR) is currently an inevitable work process in clinical hematology laboratories. The laboratory workload would be optimized if the requirement for MSR could be reduced without compromising patient care. We investigated whether slide-making rates would be different between two hematology analyzers, which were paired with their own automated slide makers/stainers: Sysmex XE-2100 with SP-1000i (Sysmex, Kobe, Japan) and ABX Pentra DX120 with SPS evolution (ABX-Horiba, Montpellier, France). METHODS: A total of 943 samples were run in parallel on the Sysmex XE-2100 and ABX Pentra DX120. Reflex slides were automatically made in each analyzer according to its own criteria, which reflected the criteria of MSR in our laboratory. The slide-making rates were compared, and the results were further confirmed using the criteria of MSR. RESULTS: The slide-making rates in Sysmex XE-2100, ABX Pentra DX120, and manual review were 22.5% (212/943), 15.91% (150/943), and 11.5% (108/943), respectively. In 774 (82.1%) samples, the three methods showed concordant results, and all made slides in 82 samples. Using the manual method as a standard, the sensitivity and specificity were 86.1% and 85.8% in Sysmex XE-2100 and 89.8% and 93.7% in ABX Pentra DX120. CONCLUSION: Our data show that the slide-making rates are variable in different hematology analyzers. It also implies that although MSR cannot be fully substituted by modern hematology analyzers, it can be effectively reduced to optimize laboratory workload.

High prevalence of autoantibodies in hepatitis A infection: the impact on laboratory profiles.

AIMS: In the absence of IgM antibodies against hepatitis A virus (HAV), HAV infections can be regarded as autoimmune hepatitis when they show positive autoantibodies by indirect immunofluorescence and lack other viral markers. The aim of this study was to evaluate the prevalence, titres and impact of autoantibodies in Korean patients with HAV infection. METHODS: The study involved a retrospective review of the electronic medical records of 73 patients with HAV at Konkuk University Hospital from August 2005 to September 2008. The presence and pattern of anti-nuclear antibody, anti-smooth muscle antibody, anti-mitochondrial antibody and anti-liver/kidney microsomal antibody were assessed by indirect immunofluorescence on Hep-2 cells and mouse/kidney sections. RESULTS: Of the 73 patients with hepatitis A, 65 (89.0%) showed positive indirect immunofluorescence tests. Of note, most of the positive tests (95.5%) showed a cytoplasmic pattern with filamentous staining of cytoplasmic fibres. There was no significant difference between groups in age or sex. In patients positive for autoantibodies, alanine aminotransferase and leucocyte count were significantly higher, while the increase in globulin was not statistically significant. In terms of titres, globulin was significantly higher in patients with > or =1:160 titres than in those with < or =1:80 titres (mean (SD) 3.4 (0.5) versus 2.8 (0.4) g/dl, respectively; p = 0.000). CONCLUSIONS: The study demonstrated a high prevalence of anticytoplasmic autoantibodies in patients with acute hepatitis A. These data would be useful to aid interpretation of indirect immunofluorescence testing in patients with acute hepatitis, especially in areas with a high prevalence of HAV.

Effect of physical exercise on renal response to head-out water immersion.

Head-out water immersion (HOI) induces various renal functional changes, such as diuresis, natriuresis, and kaliuresis. The present study was undertaken 1) to characterize the renal response to HOI in Koreans who routinely ingest high salt diet and 2) to evaluate the impact of exercise on the renal response to HOI. Six healthy male subjects (average Na+ intake of 232 mEq.day-1) were immersed upto the neck in 34.5 degrees C water and rested in a seated position or exercised on a bicycle ergometer for 3 hours. In resting subjects, we observed a reversible increase in urine flow and a decrease in urine osmolality, with no changes in creatinine clearance. The peak urine flow observed during the second hour of immersion was 4-fold greater than the pre-immersion level. The excretion of total osmotic substances rose progressively during the 3-hour immersion, which was accompanied by a similar change in Na+ excretion. The K+ excretion was slightly elevated. The major component of the immersion diuresis was a water diuresis in the early phase and an osmotic diuresis in the late phase of immersion. In exercising subjects, the diuretic and natriuretic responses to HOI were attenuated and the kaliuretic response was potentiated. Blood hemoglobin concentration and plasma levels of renin, ADH, and aldosterone decreased during immersion-rest, but they remained unchanged or increased during immersion-exercise. These results suggest that 1) the cardiac mechanoreceptor-mediated renal responses to HOI are not changed by chronic high salt diet, and 2) excessive urinary sodium and water losses are prevented by exercise during immersion.