Collaborative Study to Establish National Reference Standards for Anti-HIV-1 Antibody.

Background: National reference standards for anti-HIV-1 antibody are needed to evaluate the performance and maintain the quality control of anti-HIV-1 antibody assays. The aim of this study was to prepare a mixed-titer performance panel and assess its suitability as a national reference standard for anti-HIV-1 antibody according to stability, collaboration, and other studies. Methods: Nineteen serum samples from different HIV patients were obtained, along with 15 units of fresh frozen plasma samples with negative anti-HIV-1 antibody results. Ten anti-HIV-1 antibody-positive candidate standards and two negative candidate standards were prepared based on the reactivity in the Alinity i HIV Ag/Ab combo assay (Abbott Laboratories, Wiesbaden, Germany). A collaborative study was conducted across eight laboratories using five anti-HIV-1 antibody assays. Real-time and accelerated stability were evaluated to assess the long-term stability. Results: In the collaborative study, results of all five anti-HIV-1 antibody assays were positive for all 10 candidate standards prepared using HIV patient samples. The CV of each assay for every candidate standard was within 10%, except for one assay result. No real-time and accelerated stability change trend was observed at -70°C or -20°C, supporting that the reference standards were maintained in a stable state at -70°C for long-term storage. Conclusions: The overall results suggest that the 12 candidate standards could serve as national reference standards for anti-HIV-1 antibody.

Vivax malaria detection using a parasitic red blood cell flag generated by the Sysmex XN-9000 hematology analyzer.

INTRODUCTION: A Sysmex XN-series hematology analyzer (Sysmex), the next generation up from the Sysmex XE-series, can provide information regarding malaria infection in the form of a parasitic red blood cell (pRBC) flag. This study aimed to determine the usefulness of the pRBC flag for early detection and follow-up in patients infected with Plasmodium vivax. METHODS: A total of 221 patients with fever for whom CBC and malaria microscopy had been requested were analyzed. Sixty-seven individuals were diagnosed with P vivax infection, and 154 were diagnosed with other febrile diseases. The sensitivity and specificity of the pRBC flag for malaria parasite detection and the relationship between parasite density and presence of the pRBC flag were determined. The concordance rate between malaria microscopy and pRBC flag in 147 follow-up cases was calculated. RESULTS: The pRBC flag was detected in 56 of 67 malaria patients (sensitivity, 83.6%; specificity, 100%). The patients with the pRBC flag at initial diagnosis revealed significantly higher parasite density than the patients without the pRBC flag (P < .05). The concordance rate between malaria microscopy and pRBC flag in the follow-up cases was 53.1%. CONCLUSION: Considering its high sensitivity in malaria-suspicious patients, unexpected vivax malaria cases can be detected with the pRBC flag when CBC is done in a routine laboratory setting. The pRBC flag provided by the Sysmex XN series is a valuable tool for vivax malaria detection.

Clinical value of procalcitonin for suspected nosocomial bloodstream infection.

BACKGROUND/AIMS: Procalcitonin (PCT) may prove to be a useful marker to exclude or predict bloodstream infection (BSI). However, the ability of PCT levels to differentiate BSI from non-BSI episodes has not been evaluated in nosocomial BSI. METHODS: We retrospectively reviewed the medical records of patients ≥ 18 years of age with suspected BSI that developed more than 48 hours after admission. RESULTS: Of the 785 included patients, 105 (13.4%) had BSI episodes and 680 (86.6%) had non-BSI episodes. The median serum PCT level was elevated in patients with BSI as compared with those without BSI (0.65 ng/mL vs. 0.22 ng/mL, p = 0.001). The optimal PCT cut-off value of BSI was 0.27 ng/mL, with a corresponding sensitivity of 74.6% (95% confidence interval [CI], 66.4% to 81.7%) and a specificity of 56.5% (95% CI, 52.7% to 60.2%). The area under curve of PCT (0.692) was significantly larger than that of C-reactive protein (CRP; 0.526) or white blood cell (WBC) count (0.518). However, at the optimal cut-off value, PCT failed to predict BSI in 28 of 105 cases (26.7%). The PCT level was significantly higher in patients with an eGFR < 60 mL/min/1.73 m2 than in those with an eGFR ≥ 60 mL/min/1.73 m2 (0.68 vs. 0.17, p = 0.01). CONCLUSIONS: PCT was more useful for predicting nosocomial BSI than CRP or WBC count. However, the diagnostic accuracy of predicting BSI remains inadequate. Thus, PCT is not recommended as a single diagnostic tool to avoid taking blood cultures in the nosocomial setting.

Comparison of automated treponemal and nontreponemal test algorithms as first-line syphilis screening assays.

BACKGROUND: Automated Mediace Treponema pallidum latex agglutination (TPLA) and Mediace rapid plasma reagin (RPR) assays are used by many laboratories for syphilis diagnosis. This study compared the results of the traditional syphilis screening algorithm and a reverse algorithm using automated Mediace RPR or Mediace TPLA as first-line screening assays in subjects undergoing a health checkup. METHODS: Samples from 24,681 persons were included in this study. We routinely performed Mediace RPR and Mediace TPLA simultaneously. Results were analyzed according to both the traditional algorithm and reverse algorithm. Samples with discordant results on the reverse algorithm (e.g., positive Mediace TPLA, negative Mediace RPR) were tested with Treponema pallidum particle agglutination (TPPA). RESULTS: Among the 24,681 samples, 30 (0.1%) were found positive by traditional screening, and 190 (0.8%) by reverse screening. The identified syphilis rate and overall false-positive rate according to the traditional algorithm were lower than those according to the reverse algorithm (0.07% and 0.05% vs. 0.64% and 0.13%, respectively). A total of 173 discordant samples were tested with TPPA by using the reverse algorithm, of which 140 (80.9%) were TPPA positive. CONCLUSIONS: Despite the increased false-positive results in populations with a low prevalence of syphilis, the reverse algorithm detected 140 samples with treponemal antibody that went undetected by the traditional algorithm. The reverse algorithm using Mediace TPLA as a screening test is more sensitive for the detection of syphilis.

Colistin treatment in carbapenem-resistant Acinetobacter baumannii pneumonia patients: Incidence of nephrotoxicity and outcomes.

Colistimethate sodium (CMS) is increasingly used to treat multidrug-resistant Gram-negative bacilli infections. However, the incidence of CMS-associated nephrotoxicity has not been evaluated in patients with carbapenem-resistant Acinetobacter baumannii (CRAB) pneumonia. This retrospective study included 120 patients with CRAB pneumonia treated with intravenous CMS for ≥72 h. The objective of the study was to determine risk factors for CMS-induced nephrotoxicity and 30-day mortality in patients with CRAB pneumonia. Of the 120 patients with CRAB pneumonia, 61 (51%) developed nephrotoxicity. Multivariate analysis showed that dose per ideal body weight (IBW) [odds ratio (OR)=1.28, 95% confidence interval (CI) 1.01-1.62; P=0.04], Charlson co-morbidity index (OR=1.31, 95% CI 1.06-1.60; P=0.01) and septic shock (OR=3.16, 95% CI 1.32-7.60; P=0.01) were associated with CMS-associated nephrotoxicity. Thirty-day mortality was 33% (39/120). Multivariate analysis showed that higher daily doses of CMS per IBW [hazard ratio (HR)=0.81, 95% CI 0.67-0.98; P=0.03] and longer duration of CMS therapy (HR=0.86, 95% CI 0.79-0.95; P=0.002) were associated with increased survival. Septic shock (HR=3.91, 95% CI 1.95-7.83; P<0.001) and corticosteroid use (HR=3.49, 95% CI 1.67-7.28; P=0.001) were associated with decreased survival in patients with CRAB pneumonia. Higher daily doses of CMS per IBW, Charlson comorbidity index and septic shock were significant risk factors for CMS-associated nephrotoxicity. However, CMS-associated nephrotoxicity does not appear to have an impact on mortality.

A case of fixed drug eruption due to doxycycline and erythromycin present in food.

A fixed drug eruption (FDE) is not difficult to diagnose, given its clinical characteristics. However, the causative agent can be difficult to identify, particularly when the patient denies ingestion of any drugs. To the best of our knowledge, we present herein the first reported case of an FDE caused by antibiotics taken in food; doxycycline and erythromycin contained in pork and fish. A 57-year-old female experienced repeated episodes of well-demarcated erythematous patches covering her entire body. She denied taking any medications, but she thought that the lesions appeared after consuming pork and/or fish. An oral provocation test showed positive results for doxycycline and erythromycin, commonly used antibiotics in live-stock farming and in the fishing industry. Because of the antibiotics' thermostability, cooking does not guarantee the elimination of residual drugs. From the patient's history, we concluded that doxycycline and erythromycin contained in the pork and fish that she ate were the cause of the FDE.

Methicillin-resistant Staphylococcus aureus in nasal surveillance swabs at an intensive care unit: an evaluation of the LightCycler MRSA advanced test.

BACKGROUND: We compared the LightCycler MRSA advanced test (Roche Diagnostics, Germany) with enrichment culture methods to evaluate the relative diagnostic performance of the LightCycler MRSA advanced test for active surveillance in a high-prevalence setting. METHODS: A total of 342 nasal swab specimens were obtained from patients in the intensive care unit at admission and on the seventh day for follow-up. The results of LightCycler MRSA advanced test were compared to those of the enrichment culture. For discrepant results, mecA gene PCR was performed. RESULTS: For the detection of methicillin-resistant Staphylococcus aureus (MRSA), the LightCycler MRSA advanced test showed 98.5% sensitivity and 78.6% specificity and had positive and negative predictive values of 75.0% and 98.8%, respectively. A total of 46 samples had discrepant results between the LightCycler MRSA advanced test and enrichment culture. Of the 44 specimens that were positive in the LightCycler MRSA advanced test but negative by enrichment culture, mecA genes were detected in 37 specimens. In addition, of the original 44 cases, 21 patients had a history of MRSA colonization or infection within the last month; of those 21 specimens, 20 were positive for mecA gene as shown by PCR. Seven mecA-negative discrepant specimens comprised 3 methicillin-sensitive S. aureus-culture positive and only 2 patients had MRSA infections. CONCLUSIONS: Despite its low specificity and positive predictive value, the LightCycler MRSA advanced test could serve as a rapid test for patients colonized with MRSA.

Association between changes in lipid profiles and progression of symptomatic intracranial atherosclerotic stenosis: a prospective multicenter study.

BACKGROUND AND PURPOSE: Predictors of progression of intracranial atherosclerotic stenosis have not been clearly identified. We investigated whether poststroke changes in lipid profiles would affect the prognosis of symptomatic intracranial atherosclerotic stenosis. METHODS: This is a substudy of Trial of cilOstazol in Symptomatic intracranial Stenosis 2 (TOSS-2). From 10 centers we enrolled 230 subjects with acute symptomatic stenosis in the M1 segment of the middle cerebral artery or basilar artery. At baseline and 7 months after stroke, subjects underwent MR angiogram and assessment of cardiovascular risk factors including lipoprotein levels. Progression of intracranial atherosclerotic stenosis was determined by comparing stenosis on the baseline and follow-up MR angiograms. RESULTS: Cilostazol treatment was more frequently seen in the nonprogression group (109 of 198 [55.1%]) than in the progression group (11 of 32 [34.4%]). At 7 months after stroke when compared with baseline, low-density lipoprotein cholesterol and total cholesterol levels decreased in both groups. However, only nonprogressors showed increase in high-density lipoprotein cholesterol levels between baseline and follow-up. Changes in apolipoprotein B/apolipoprotein A-I levels were not different between the groups, although apolipoprotein B/A-I at 7 months was higher in progressors than in nonprogressors. Remnant lipoprotein cholesterol levels decreased in nonprogressors, whereas they did not change in progressors. In multivariable analyses, after adjusting for cilostazol treatment and remnant lipoprotein cholesterol reduction or apolipoprotein B/A-I at 7 months, high-density lipoprotein cholesterol elevation remained as a significant predictor for the nonprogression. CONCLUSIONS: This is the first prospective multicenter study to demonstrate that high-density lipoprotein cholesterol elevation, along with remnant lipoprotein cholesterol reduction and low apolipoprotein B/A-I, is associated with prevention of angiographic progression of symptomatic intracranial atherosclerotic stenosis. CLINICAL TRIAL REGISTRATION INFORMATION: URL: http://www.clinicaltrials.gov. Unique identifier: NCT00130039.

A case of monoclonal gammopathy in extranodal marginal zone B-cell lymphoma of the small intestine.

Monoclonal gammopathy occurs in one-third of the patients with mucosa-associated lymphoid tissue lymphoma (MALT lymphoma). However, monoclonal gammopathy has been rarely reported in Korea. Paraprotenemia accompanying MALT lymphoma is strongly correlated with involvement of the bone marrow, and this involvement leads to the progression of the disease. Here, we present a case of a 66-yr-old man diagnosed with IgM monoclonal gammopathy and stage IV extranodal marginal zone lymphoma of the small intestine, with the involvement of the bone marrow.

High levels of remnant lipoprotein cholesterol is a risk factor for large artery atherosclerotic stroke.

BACKGROUND AND PURPOSE: Remnant lipoproteins (RLPs) are products of partially catabolized chylomicrons and very-low-density lipoprotein, from which some triglycerides have been removed. These particles are smaller and denser than the parent particles and are believed to be strongly atherogenic. We explored the association between RLP cholesterol (RLP-C) and ischemic stroke, including stroke subtypes. METHODS: A cohort of 142 ischemic stroke patients (90 men and 52 women; age, 65.2±12.8 years, mean±SD) was enrolled; all had acute infarcts confirmed by diffusion-weighted MRI, and had fasting lipograms. A full stroke-related evaluation was conducted on each patient. An outpatient population of 88 subjects without a history of cerebrovascular or cardiovascular disease served as a control group. Serum RLP fractions were isolated using an immunoaffinity gel containing specific antiapolipoprotein (anti-apo)B-100 and anti-apoA-I antibodies. RLP-C values were considered to be high when they were in the highest quartile of all values in the study. RESULTS: High RLP-C values were more common in stroke patients than in control patients (31.0% vs. 14.8%, p=0.01), when 5.6 mg/dL (>75th percentile) was used as the cutoff value. Multivariable analyses indicated that RLP-C was a risk factor for stroke, with an odds ratio of 2.54 (p=0.045). The RLP-C level was higher in the large artery atherosclerosis subgroup (5.7±3.9 mg/dL) than in any other stroke subgroup (small vessel occlusion, 4.9±5.9 mg/dL; cardioembolism, 1.8±2.3 mg/dL; stroke of undetermined etiology, 3.1±2.9 mg/dL). CONCLUSIONS: We have found an association between high RLP-C levels and ischemic stroke, and in particular large artery atherosclerotic stroke.

[Current status and proposal of a guideline for manual slide review of automated complete blood cell count and white blood cell differential].

BACKGROUND: Manual slide review (MSR) is usually triggered by the results of automated hematology analyzers, but each laboratory has different criteria for MSR. This study was carried out to investigate the current status of MSR criteria of automated complete blood cell count (CBC) and white blood cell (WBC) differential results and to propose a basic guideline for MSR. METHODS: Total 111 laboratories were surveyed regarding MSR using questionnaires. The questionnaire asked: kinds of automated hematology analyzers used and the presence of criteria triggering MSR in seven categories: 1) CBC results, 2) 5 differential WBC counts, 3) 3 differential WBC counts, 4) automated reticulocyte counts, 5) delta check, 6) instrument flags (or messages), 7) clinical information (wards or diseases). Based on the survey results, we determined basic and extended criteria for MSR. With these criteria, we consulted nine hematology experts to get a consensus. RESULTS: All 111 laboratories had their own MSR criteria. Among 111 laboratories, 98 (88.3%) used more than three criteria for MSR including CBC results and 5-part WBC differential count results and 95 (85.6%) had criteria of flags triggering MSR. For MSR criteria with numeric values, the 10th, 50th, and 90th percentiles of upper and lower threshold values were obtained. The basic guideline for MSR was made. CONCLUSIONS: We proposed a basic guideline for MSR. This guideline would be helpful to hematology laboratories for their daily operation and providing more rapid and accurate CBC and WBC differential results.

[Current status of external quality assessment of fecal occult blood test].

BACKGROUND: Nationwide external quality assessment (EQA) of the fecal occult blood test (FOBT) in Korea was first introduced in 2007-2009. The EQA results were analyzed to assess the current status of FOBT and to plan the continuation of the EQA program. METHODS: The surveys included 40 hospitals in the preliminary survey conducted in 2007, 249 general hospitals in 2008, and 389 hospitals in 2009. In the surveys, the participating hospitals provided the results of the distributed materials and replies to the questionnaire on the FOBT test procedures and quality controls. RESULTS: In the surveys conducted between 2007 and 2009, a total of 650 institutes submitted 653 test system results; 3 institutes used 2 kinds of methods. All of the institutes used immunologic methods; 107 institutes (16.5%) used quantitative equipments and 546 institutes (84.0%) used qualitative kits. Most quantitative tests yielded consistent positive or negative results; however, their cut-off and measured values differed according to the equipments used. A low-level material tested in 2007 was negative in the quantitative methods but positive in some qualitative methods because of lower detection limits. The discordance rates among quantitative tests were 3.2% in 2007, 4.4% in 2008, and 0% in 2009 and the rates among qualitative tests were 13.8% in 2008 and 2.6% in 2009. Semi-solid EQA materials showed the ability to evaluate the overall test procedures with acceptable stability. CONCLUSIONS: In the first Korean FOBT EQA, commercially available EQA materials were proven to be stable. Continuation of the EQA program and further education of laboratory personnel are needed to reduce inconsistency in results. Further, the test kit, procedures, and result reports must be standardized.

[A case of post-essential thrombocythemia myelofibrosis with severe osteosclerosis].

Essential thrombocythemia (ET) is a chronic myeloproliferative neoplasm that involves primarily the megakaryocytic lineage. After many years, a few patients with ET may develop bone marrow (BM) fibrosis and rarely develop osteosclerosis. A 60-yr-old female was admitted due to severe left upper quadrant abdominal discomfort. She had been diagnosed as ET 19 yrs ago. On liver computed tomography severe splenomegaly was shown. Laboratory tests revealed WBC 24.3x10(9)/L, hemoglobin 13.4 g/dL, platelets 432x10(9)/L, lactate dehydrogenase 4,065 IU/L (reference range; 240-480). Blood smear demonstrated leukoerythroblastosis, teardrop cells, and giant and hypogranular platelets. BM study revealed inadequate aspirate due to dry tap. BM biopsy showed clusters of dysplastic megakaryocytes, grade 3 fibrosis, and severe osteosclerosis. Major/minor BCR-ABL1 rearrangement and JAK2 V617F mutation were not detected. Cytogenetic studies revealed normal karyotype. According to the 2008 WHO diagnostic criteria, the patient was diagnosed as having post-essential thrombocythemia myelofibrosis with severe osteosclerosis.

[Evaluation of the BD phoenix automated microbiology system SMIC/ID-2 panel for antimicrobial susceptibility testing of Streptococcus pneumoniae].

BACKGROUND: With the emergence of antimicrobial resistance among Streptococcus pneumoniae, a more accurate and automated antimicrobial susceptibility testing method is essential. We evaluated the BD Phoenix Automated Microbiology System (Becton Dickinson Diagnostic Systems, USA) SMIC/ID-2 panel for antimicrobial susceptibility testing of S. pneumoniae. METHODS: A total of 113 clinical strains of S. pneumoniae (88 penicillin susceptible strains, 8 intermediate strains, and 17 resistant strains by 2008 CLSI criteria) were tested. Minimum inhibitory concentrations (MICs) for penicillin, cefotaxime, clindamycin, erythromycin, levofloxacin, trimethoprim/ sulfamethoxazole, tetracycline, and vancomycin were determined by Etest (AB Biodisk, Sweden) and Phoenix System. The results obtained by Phoenix system were compared to those obtained by Etest. RESULTS: The overall essential agreement of MICs (within one dilution of MICs) defined by the Phoenix and Etest was 92.3%. Neither very major errors nor major errors were produced, and minor errors were 6.5%. Minor errors were frequently observed in susceptibility testings for penicillin (22.1%), cefotaxime (12.4%), and trimethoprim/sulfamethoxazole (11.5%). CONCLUSIONS: The Phoenix SMIC/ID-2 panel provided a simple and rapid susceptibility testing for S. pneumoniae, and the results were in a good agreement with those of Etest. The Phoenix system appears to be an effective automated system in clinical microbiology laboratories.

The implementation and effects of a clinical laboratory accreditation program in Korea from 1999 to 2006.

BACKGROUND: The Korean Laboratory Accreditation Program (KLAP) by the Korean Society of Laboratory Medicine (KSLM) was started in 1999. We summarized history and achievement of KLAP for the last 8 yr. METHODS: We analyzed 8 yr data (1999-2006) of historical events, trends of participating laboratories, and scores according to the impact of the question to the outcome of the tests. Inspection check lists are for 'laboratory management', 'clinical chemistry', 'diagnostic hematology', 'clinical microbiology', 'diagnostic immunology', 'transfusion medicine', 'cytogenetics', 'molecular genetics', 'histocompatibility', 'flow cytometry', and 'comprehensive laboratory test verification report'. The laboratories with score 90 or higher got 2-yr certificate and laboratories with score between 60 and 89 got 1-yr certificate. The laboratories with score below 60 failed accreditation. RESULTS: The number of accredited laboratories was 2.4 times higher in 2006 (n=227) than in 1999 (n=96). Inspection check lists have been revised 5 times till 2006. The average accreditation rate was 99.6% during these periods and the 2-yr accreditation rate was 32.4% in 2000, 45.6% in 2001, 53.3% in 2002, 47.3% in 2003, 68.5% in 2004, 37.7% in 2005, and 47.7% in 2006. Number of participants in inspector training workshops increased from 89 in 2000 to 766 in 2006. CONCLUSIONS: The KLAP has been in place successfully and stabilized over the past 8 yr. It seemed to enhance the laboratory quality. Efforts for improvement of quality control and inspector training workshops appeared to be in the main contributing factors.

A case of myocarditis associated with Plasmodium vivax malaria.

Cardiac complications in malaria have been infrequently associated with Plasmodium falciparum infections. However, myocarditis associated with Plasmodium vivax malaria has not been reported in the literature. We observed an unusual case of vivax malaria that was complicated by myocarditis.

[Effectiveness of sodium fluoride as a glycolysis inhibitor on blood glucose measurement: comparison of blood glucose using specimens from the Korea National Health and Nutrition Examination survey].

BACKGROUND: Accurate measurement of blood glucose concentrations is essential for defining diabetes, and the minimization of ex vivo glycolysis has been recommended. Recent guidelines advocate two kinds of methods for sample collection and processing: either the sodium fluoride (NaF) method or immediate refrigeration using a serum separation tube (SST). We investigated the difference between the two methods in measuring subsequent glucose concentrations using blood specimens from participants recruited for the fourth Korean National Health and Nutrition Examination Survey. METHODS: Paired venous blood samples were collected in an SST and a NaF tube from 1,103 men and women. SST serum was separated within 30 min, including standing for 15 min, and then refrigerated. The NaF samples were refrigerated, but not separated until immediately before analysis. We compared the blood glucose concentrations between the SST (SST glucose) and NaF (NaF glucose) methods. RESULTS: The mean SST glucose was significantly higher than NaF glucose (99.0 mg/dL vs 96.5 mg/dL, P<0.05). NaF glucose showed a negative mean bias of 2.6 mg/dL vs SST glucose but showed high correlation (R=0.9899). There was no significant correlation between the bias of blood glucose concentrations by two methods and the storage time of NaF glucose. CONCLUSIONS: The negative bias associated with the use of NaF tubes may significantly affect the prevalence of diabetes. Serum separation and refrigeration within 30 min after venous sampling is recommended over NaF method, not only to minimize the preanalytical impact on detecting diabetes but also to reduce sample volume and number of tubes.