Il Joong Park

VanB phenotype-vanA genotype Enterococcus faecium with heterogeneous expression of teicoplanin resistance.

ABSTRACT Six VanB phenotype-vanA genotype isolates of Enterococcus faecium with heterogeneous expression of teicoplanin resistance which gave rise to an outbreak at a Korean tertiary care teaching hospital have IS1216V in the coding region of vanS. This could be the underlying cause of the VanB phenotype-vanA genotype with heterogeneous expression of teicoplanin resistance.

Acute myeloid leukemia with t(16;21)(q24;q22) and eosinophilia: case report and review of the literature

The t(16;21)(q24;q22), a rare chromosomal translocation involving chromosome 21 in de novo and therapy-related acute myeloid leukemia (AML), produces a RUNX1-CBFA2T3 fusion gene (previously AML1-MTG16) fusion gene. The translocation has been reported in 20 patients with AML, with eosinophilia present in 3 cases. Here we report a pediatric case of t(16;21)(q24;q22) in de novo AML with eosinophilia and suggest that eosinophilia is a hematologic characteristic of at least a subpopulation of AML with t(16;21)(q24;q22). A 4-year-old Korean girl was admitted with complaints of pale appearance and dizziness, and was diagnosed with acute myelomonocytic leukemia. On admission, laboratory evaluation revealed hemoglobin at 3.3 g/dL, platelets at 9.0 x 10(9)/L, and white blood cells at 9.1 x 10(9)/L with 10% eosinophils and 1% blasts. The bone marrow aspirate contained 31% blasts and 11% eosinophils. Flow cytometric analysis revealed the expression of CD13, CD14, CD19, CD33, CD34, and HLA-DR by the leukemic blasts. The karyotype was 47,XX, + 8,t(16;21)(q24;q22)[18]/46,XX[2]. Interphase fluorescence in situ hybridization analysis with a dual-color, dual-fusion translocation LSI AML1/ETO probe set for RUNX1 and RUNX1T1 produced three signals for each probe in 90% of interphases, but no fusion signals. We confirmed the presence of RUNX1-CBFA2T3 fusion transcripts with reverse transcriptase-polymerase chain reaction, using primers AML1ex5f1 and MTG16r2.

Characterization of a Vancomycin-resistant Enterococcus faecium Outbreak Caused by 2 Genetically Different Clones at a Neonatal Intensive Care Unit

In July 2010, we identified an outbreak of vancomycin-resistant enterococci (VRE) in our 26-bed neonatal intensive care unit. We performed an epidemiological investigation after clinical cultures of 2 neonates were positive for VRE. Identification, susceptibility testing, and molecular characterization were performed. Cultures of 3 surveillance stool samples of inpatients and 5 environmental samples were positive for VRE. All isolates were identified as Enterococcus faecium containing the vanA gene. Two distinct clones were identified by performing pulsed-field gel electrophoresis. The 2 clones exhibited different pulsotypes, but they represented identical Tn1546 types. Two sequence types, ST18 and ST192, were identified among all of the isolates with multilocus sequence typing. Our investigation determined that the outbreak in the neonatal intensive care unit was caused by 2 genetically different clones. The outbreak may have occurred through clonal spread and horizontal transfer of the van gene.

Genetic Rearrangements of Tn1546-Like Elements in Vancomycin-Resistant Enterococcus faecium Isolates Collected from Hospitalized Patients over a Seven-Year Period

ABSTRACT The heterogeneity of Tn1546 results from point mutations, deletions, and the integration of insertion sequence (IS) elements. Among these variations, the presence of IS elements accounts for much of the heterogeneity. Such a rearrangement could play a key role in the evolution of the vanA gene cluster, and hence, it may modify its transferability. In this study, we characterized the consequence of Tn1546 in vanA-containing Enterococcus faecium isolates collected from patients over time. From 1998 to 2004, 57 vanA-containing E. faecium isolates were collected from hospitalized patients at Ajou University Hospital in Korea. PCR amplification of internal regions of Tn1546 was performed, and both DNA strands were directly sequenced by the dideoxy termination method. All isolates were divided into three main types, including the prototype, according to the distribution of IS elements integrated into Tn1546 elements. Type I was characterized by an IS1542 insertion in the orf2-vanR intergenic region and an IS1216V insertion in the vanX-vanY intergenic region. Type II was represented by the presence of two copies of IS1216V at the 3′ end of IS1542 and in the vanX-vanY intergenic region, as well as IS1542 in the orf2-vanR intergenic region. Seventeen strains isolated from 1998 to 2000 represented type I, and 38 strains isolated from 2000 to 2004 represented type II. The remaining two isolates were the prototype. The tendency for the rearrangement of Tn1546 was that the sequences were shortened as time passed, especially at the left or the right end, and hence, this could gradually modulate their transferability.

A case of chronic myelogenous leukemia with e8a2 fusion transcript

The Philadelphia chromosome and its corresponding fusion gene, BCR-ABL, is one of the best-known genetic abnormalities in hematological malignancies. Major BCR-ABL translocation is much more common in chronic myelogenous leukemia (CML) and minor BCR-ABL in acute lymphoblastic leukemia. We experienced an extraordinarily rare case of CML with an e8a2 variant. An unusual band, other than the common transcripts, was observed in reverse transcription-polymerase chain reaction (RT-PCR) for the BCR-ABL gene rearrangement. Sequence analysis of the PCR product revealed an 1172-bp e8a2 fusion with a 14-bp insertion of ABL intron Ia. The patient achieved a complete hematological response 3 months after imatinib treatment. It is necessary to keep in mind that an unexpected band revealed with RT-PCR may mean the presence of unusual fusion gene.

Relapse and reacquisition of rectal colonization by vancomycin-resistant Enterococcus faecium after decolonization.

To better understand the epidemiology of colonization of vancomycin-resistant enterococci (VRE), we performed an 8-year retrospective study of all hospitalized patients with recurrent VRE colonization after they were documented as being clear of VRE and compared the primary colonization isolates and recolonization isolates by pulsed-field gel electrophoresis and Tn1546 typing. Review of the medical records of all patients showed that of the 15 patients with recurrent colonization, six continued to be hospitalized on the same floor. Five were discharged home and then readmitted. Four were moved to another floor. Patients who remained on the same floor were recolonized with a strain that was indistinguishable from the original colonizing strain. Patients who were moved or were discharged had de novo VRE colonization with strains distinct from the original colonizing strain.

[Two cases of acute myeloid leukemia with t(16;21)(p11;q22) and TLS/FUS-ERG fusion transcripts].

Many AML-associated chromosomal abnormalities, such as t(8;21), t(15;17), inv(16), t(9;11), t(9;22) and t(6;9) are well known. The chromosomal aberration of t(16;21)(p11;q22) in AML is rare and it is known to be associated with poor prognosis, young age (median age, 22 yr), and involvement of various subtypes of the French-American-British classification. We report here 2 AML patients with t(16;21)(p11;q22), proved by conventional cytogenetics and/or reverse transcription (RT)-PCR. Erythrophagocytosis by leukemic blasts was observed in both of the cases. One patient was a 24 yr-old male with acute myelomonocytic leukemia. His karyotype was 46,XY,t(16;21)(p11;q22),del(18)(p11.2) and RT-PCR revealed the TLS/FUS-ERG fusion transcripts. Although he received allogeneic peripheral blood stem cell transplantation after the first remission, he died 9 months after the initial diagnosis due to relapse of the disease and graft-versus-host disease. The other patient was a 72 yr-old male with acute myeloid leukemia without maturation. His karyotype was 45,XY,-16,add(21)(q22) and the presence of t(16;21)(p11;q22) was detected by RT-PCR. He was transferred to another hospital with no more follow-up. We suggest that the presence of t(16;21)(p11;q22) and/or TLS/FUS-ERG fusion transcripts has to be considered in cases of AML with erythrophagocytosis.

Usefulness of delta value of platelet parameters on ADVIA 120 for the functional reactivity of stored platelets.

An agonist‐induced expression of CD62P by flow cytometry analysis for evaluating platelet functional reactivity has some disadvantages. We investigated the usefulness of platelet parameters by ADVIA 120 to predict an agonist‐induced expression of CD62P in stored platelets. The CD62P expression by flow cytometry and the platelet parameters by ADVIA 120 were studied in samples from 27 platelet pheresis products. Delta (Δ) values were calculated as the degree of change of the platelet parameters studied with or without adenosine 5′‐diphosphate sodium (ADP) stimulation. The CD62P expression of the ADP‐activated platelets were correlated with the Δplatelet count (r=0.517) in the short‐term storage group (within 10 hr from preparation), with the platelet component distribution width (PCDW) without ADP (r=−0.744) and the ΔPCDW (r=−0.755) in the long‐term storage group (after 10 hr from preparation). Therefore, the delta values of platelet parameters on ADVIA 120 analysis in platelets between with and without ADP stimulation could be useful as a simple predictor for the functional reactivity of stored platelets. J. Clin. Lab. Anal. 24:38–43, 2010.

Frequency, interobserver reproducibility and clinical significance of equivocal peaks in PCR clonality testing using Euroclonality/BIOMED-2 primers

Aims PCR studies for lymphoid clonality are now widely employed, especially using Euroclonality/BIOMED-2 primers. Criteria for interpretation as a clonal result, however, have proven controversial. This study examines the frequency and clinical significance of equivocal amplification patterns and measures the interobserver reproducibility of clonality interpretations. Methods At our institution, results of each primer set are first classified as clonal, non-clonal or abnormal (equivocal peak on polyclonal background). Final results for all primer sets are then collectively reported as positive (≥1 clonal result), negative (non-clonal results) or indeterminate (≥1 abnormal result) for a clonal population. Results of 274 consecutive clonality cases were reviewed, and the interobserver reproducibility of individual primer set reactions and final results was determined in a subset of 30 cases. Results 44/161 (27%) B-cell and 50/163 (31%) T-cell cases contained at least one abnormal peak. Of these, 29 (64%) and 31 (62%), respectively, showed clonal results in another primer set. Interobserver reproducibility was excellent for most primer sets and for final interpretations, but only fair to good for IGK V-J and TCRB D-J1+2 primer sets. A definitive diagnosis of lymphoma was rendered in 93%, 20% and 6% of B-cell cases and 90%, 42%, and 14% of T-cell cases positive, indeterminate or negative for a clonal population, respectively. Conclusions Using a subjective approach, abnormal (equivocal) peaks are frequently observed in routine practice. However, most cases with abnormal peaks contain clonal rearrangements in other primer sets, facilitating overall interpretation of final results with excellent interobserver reproducibility.

A Case of Escherichia coli O157 Hemorrhagic Colitis

Escherichia coli O157 is an important serotype of enterohemorrhagic E. coli that causes hemorrhagic colitis worldwide. Outbreaks of E. coli O157 have been assocoated with contaminated food like meat, raw milk, and water, but recently vegetables and fruits have accounted for a growing number of recognized outbreaks. We isolated verotoxin producing E. coli O157 from the stool of a 3 year-old female with bloody diarrhea and abdominal pain. The child had been eating salad with vegetables and fruits frequently. (Korean J Clin Microbiol 2008;11:66-68)