Hee Jin Huh

Prognostic significance of multidrug resistance gene 1 (MDR1), multidrug resistance-related protein (MRP) and lung resistance protein (LRP) mRNA expression in acute leukemia.

The prognostic significance of multidrug resistance (MDR) gene expression is controversial. We investigated whether multidrug resistance gene 1 (MDR1), multidrug resistance-related protein (MRP) and lung resistance protein (LRP) mRNA expression are associated with outcomes in acute leukemia patients. At diagnosis we examined MDR1, MRP and LRP mRNA expression in bone marrow samples from 71 acute leukemia patients (39 myeloid, 32 lymphoblastic) using nested RT-PCR. The expression of each of these genes was then expressed as a ratio in relation to β-actin gene expression, and the three genes were categorized as being either 0, 1+, 2+ or 3+. MDR1, MRP and LRP mRNA expression was detected in 23.9%, 83.1% and 45.1%, respectively. LRP mRNA expression was significantly associated with resistance to induction chemotherapy in acute leukemia patients, and in the AML proportion (p=0.02 and p=0.03, respectively). MRP and high MDR1 mRNA expression was associated with poorer 2-yr survival (p=0.049 and p=0.04, respectively). Patients expressing both MRP and LRP mRNA had poorer outcomes and had worse 2-yr survival. The present data suggest that MDR expression affects complete remission and survival rates in acute leukemia patients. Thus, determination of MDR gene expression at diagnosis appears likely to provide useful prognostic information for acute leukemia patients.

Malaria detection with the Sysmex XE-2100 hematology analyzer using pseudoeosinophilia and abnormal WBC scattergram

Recent investigation using the Sysmex XE-2100 hematology analyzer (Sysmex Corporation, Japan) has demonstrated erroneously high eosinophil counts and abnormal white blood cell (WBC) scattergrams in malaria cases. This study was conducted to assess the diagnostic efficiency of the Sysmex XE-2100 analyzer for malaria. One hundred forty-four patients initially diagnosed with Plasmodium vivax infection, 319 patients with febrile illness, and 24 patients who underwent malaria treatment were analyzed. Atypical features on Sysmex XE-2100 analyzer were categorized as pseudoeosinophilia (a gap of more than 5% in eosinophil counts between the Sysmex XE-2100 analyzer and microscopic examination) and abnormal WBC scattergram. Pseudoeosinophilia or abnormal WBC scattergram were detected in 100 of 144 malaria-positive samples (sensitivity 69.4%, specificity 100%). The samples with pseudoeosinophilia or abnormal WBC scattergrams showed significantly higher parasite counts than the samples without pseudoeosinophilia or an abnormal WBC scattergram (P < 0.05). All 24 samples from patients for whom the malaria smear was repeated after malaria treatment was initiated showed a normalized eosinophil count and a normal WBC histogram. In conclusion, attention to differential count and WBC scattergrams provided by the Sysmex XE-2100 would be a valuable tool in malaria detection.

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.

Secondary syphilis presenting as a generalized lymphadenopathy: clinical mimicry of malignant lymphoma.

The diagnosis of syphilis remains challenging. The absence of classical features of the disease, such as the rash of secondary syphilis or genital lesion, may pose diagnostic difficulties. In this article, we report a case of secondary syphilis in which the clinical syndrome and pattern of fluorodeoxyglucose uptake mimicked malignant lymphoma. This case highlights the importance of thorough history taking including sexual contact. Clinicians should be alert for syphilis-underlying unexplained lymphadenopathy, even in the absence of typical rash or genital lesion.

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.

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.

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.

Evaluation of the Bio-Rad Geenius HIV 1/2 Confirmation Assay as an Alternative to Western Blot in the Korean Population: A Multi-Center Study.

Recently updated recommendations for diagnosis of HIV infection suggest a new diagnostic algorithm including HIV-1/HIV-2 antibody differentiation immunoassay instead of western blot (WB) as a confirmatory testing. We evaluated Bio-Rad Geenius HIV1/2 confirmation assay as a simple and reliable alternative to WB in the Korean population with low HIV prevalence. The Geenius HIV1/2 was performed in a total of 192 serum specimens (140 reactive and 52 nonreactive specimens by ARCHITECT HIV Ag/Ab Combo assay) that were prospectively collected from five institutions. HIV-1 nucleic acid amplification test (NAT) was performed in negative or indeterminate specimens by Geenius HIV1/2 or WB. Among 140 reactive specimens by HIV Ag/Ab assay, 82 (58.6%) were positive for HIV-1 Ab by Geenius HIV1/2. Among 58 negative or indeterminate specimens by Geenius HIV1/2, four specimens (6.9%) were positive by HIV-1 NAT. The sensitivity and specificity of Geenius HIV1/2 were 95.3% and 100.0%, respectively. When we considered only WB, the sensitivity and specificity of Geenius HIV1/2 were 100.0% and 99.1%, respectively. Agreement between Geenius HIV1/2 and WB was excellent (weighted Kappa = 0.89). The Geenius HIV1/2 is simple and time-saving compared with WB. It has an excellent performance and can be a reliable alternative to WB. HIV-1 NAT should be performed in negative or indeterminate specimens by Geenius HIV1/2 to detect acute HIV infection as recommended in new HIV testing algorithms.

Evaluation of the Phoenix Automated Microbiology System for Detecting Extended-Spectrum β-Lactamase in Escherichia coli, Klebsiella species and Proteus mirabilis

BACKGROUND The aim of this study was to compare the BD Phoenix (Beckton Dickinson Diagnostic Systems, USA) extended-spectrum beta-lactamase (ESBL) test with the Clinical and Laboratory Standards Institute (CLSI) ESBL phenotypic confirmatory test by disk diffusion (CLSI ESBL test) in Escherichia coli, Klebsiella pneumoniae, Klebsiella oxytoca and Proteus mirabilis. METHODS We tested 224 clinical isolates of E. coli, K. pneumoniae, K. oxytoca and P. mirabilis during May 2006 to March 2007. These isolates were examined by the Phoenix and the CLSI ESBL tests simultaneously. For the isolates showing discordant results between the two tests, boronic acid disk test was performed to differentiate AmpC beta-lactamase and ESBL. RESULTS Among the 224 clinical isolates, 75 and 79 isolates were positive for ESBL by CLSI ESBL test and Phoenix test, respectively. Having detected 4 more isolates as ESBL-producers, Phoenix test showed a 98.2% agreement with a 100% sensitivity and 97.3% specificity compared with CLSI ESBL test. Among the four false positive isolates, three were AmpC-positive but ESBL-negative. CONCLUSIONS The BD Phoenix ESBL test was sensitive and specific, and can be used as a rapid and reliable method to detect ESBL production in E. coli, Klebsiella species, and P. mirabilis.

Investigation of bone marrow involvement in malignant lymphoma using fluorescence in situ hybridization: possible utility in the detection of micrometastasis

We evaluated the usefulness of interphase fluorescence in situ hybridization (FISH) for the detection of bone marrow involvement of lymphoma, comparing the results with those of microscopic examination. Bone marrow aspirates obtained for staging work-up from 150 patients with non-Hodgkin lymphoma were used in this study. Interphase FISH study using four probes and conventional G-banding were performed on bone marrow aspirates. The four probes included locus specific identifier (LSI) immunoglobulin heavy chain (IGH) dual-color break-apart rearrangement probe, an LSI p16 SpectrumOrange/CEP 9 SpectrumGreen probe, an LSI BCL6 dual-color break-apart rearrangement probe. Among 150 cases, 29 cases (19.3%) showed infiltration of neoplastic lymphoid cells by microscopic examination. Chromosomal aberrations were detected by FISH in eight patients and by conventional cytogenetic study in three patients. FISH study showed 14q32 rearrangement in four patients (4/126, 3.2%), 9q21 rearrangement in no patients (0/144, 0%), 3q27 rearrangement in four patients (4/131. 3.1%), and a gain of 1q21q32 in two patients (2/115, 1.7%). Among eight patients with abnormal FISH patterns, six had normal karyotypes or no analyzable metaphase according to the conventional cytogenetic study. Seven patients with FISH abnormality showed bone marrow involvement of lymphoma by microscopic examination. One patient, who was defined as having no evidence of bone marrow involvement by microscopic examination, showed a 3q27 aberration in the FISH study. Although the number of patients with BM involvement that was detected by FISH was low, abnormal FISH patterns were detected in six patients who did not have abnormal karyotypes. Therefore, FISH analysis would be beneficial in cytogenetic diagnosis and follow-up study of minimal residual diseases, once the cytogenetic changes are detected at initial diagnosis.