Abstract: Human epididymis protein 4 (HE4) is a new biomarker for the detection of ovarian cancer. We evaluated the analytical performance of a novel automated HE4 assay and established reference ranges of HE4 and CA125. We also compared the diagnostic performance of both biomarkers for ovarian cancer. Precision performances and linearity of the HE4 assay were assessed. Serum samples from 2,182 healthy and 72 pregnant women were also assayed for HE4 and CA125, and the 95%, 97.5% and 99% reference limits for both markers were calculated. Additionally, sera from 66 ovarian cancer and 257 benign gynecologic disease patients were tested to validate reference ranges and diagnostic performances. The total precision of the HE4 assay was <5% coefficient of variation for most of the levels evaluated. The linearity range of this assay was from 15.0 to 1100.0 pmol/L. The 97.5% upper reference limits for HE4 and CA125 were 33.2 pmol/L (95% confidence interval [CI], 32.2-34.0) and 38.3 U/mL (95% CI, 35.1-41.5), respectively. Using these values as cutoff points, the sensitivity and specificity of HE4 for differentiating ovarian cancer from benign gynecologic diseases and healthy individuals were 90.9% and 94.1%, and those of CA125 were 72.7% and 94.4%. The receiver operating characteristic-area under the curve values of HE4 and CA125 for discriminating ovarian cancer from age-matched control were 0.94 and 0.86, respectively, and they were statistically different (p = 0.0095). The new automated HE4 assay showed good analytical and diagnostic performances. The reference limits established in our study could be used as cutoff levels to facilitate more accurate diagnosis of ovarian cancer in Asian population.
Abstract: The hepatitis B virus (HBV) PreS mutations C1653T, T1753V, and A1762T/G1764A were reported as a strong risk factor of hepatocellular carcinoma (HCC) in a meta-analysis. HBV core promoter overlaps partially with HBx coding sequence, so the nucleotide 1762 and 1764 mutations induce HBV X protein (HBx) 130 and 131 substitutions. We sought to elucidate the impact of HBx mutations on HCC development. Chronically HBV-infected patients were enrolled in this study: 42 chronic hepatitis B (CHB) patients, 23 liver cirrhosis (LC) patients, and 31 HCC patients. Direct sequencing showed HBx131, HBx130, HBx5, HBx94, and HBx38 amino acid mutations were common in HCC patients. Of various mutations, HBx130+HBx131 (double) mutations and HBx5+HBx130+HBx131 (triple) mutations were significantly high in HCC patients. Double and triple mutations increased the risk for HCC by 3.75-fold (95% confidence interval [CI] = 1.101 to 12.768, P = 0.033) and 5.34-fold (95% CI = 1.65 to 17.309, P = 0.005), respectively, when HCC patients were compared to CHB patients. Functionally, there were significantly higher levels of NF-κB activity in cells with the HBx5 mutant and with the double mutants than that of wild-type cells and the triple-mutant cells. The triple mutation did not increase NF-κB activity. Other regulatory pathways seem to exist for NF-κB activation. In conclusion, a specific HBx mutation may contribute to HCC development by activating NF-κB activity. The HBx5 mutation in genotype C2 HBV appears to be a risk factor for the development of HCC and may be used to predict the clinical outcomes of patients with chronic HBV infection.
Abstract: Some tumor markers, including carcinoembryonic antigen (CEA) and cytokeratin 19 fragment (CYFRA 21-1), are used for the detection of lung cancer; however, their use is limited because of low sensitivities and high false-positive rates.
Abstract: to investigate the epidemiological traits of carbapenem-non-susceptible Acinetobacter baumannii (CNSAB) and the usefulness of phylogenetic grouping based on partial rpoB gene sequencing in defining the epidemiological traits of CNSAB.
Abstract: Interpretation of indeterminate results by anti-HIV Western blot assay, which is currently used as a confirmatory test for HIV infection, can be usually difficult. We analyzed outcomes of the patients with indeterminate results by anti-HIV Western blot. Medical records of patients, who were indeterminate by the anti-HIV Western blot assay in a university hospital during recent 5 years, were retrospectively reviewed. HIV screening test was performed by chemiluminescent immunoassay autoanalyzer (Abbot Laboratories, USA) with HIV Ag/Ab Combo kits. Confirmatory Western blot assay for the positive samples by HIV screening test was committed to the Korean National Institute of Health. A total of 202,639 specimens were tested for HIV screening during the period, and 644 (0.32%) sera showed positive results. Among these, 46 (7.1%) cases were indeterminate by the Western blot, which were from 20 patients, and 13 of them converted to be anti-HIV positive, and 3 were lost to follow-up. Another four patients were turned out to be negative for HIV infection, including two neonates from HIV-positive mothers receiving antiviral treatment during pregnancy. Most of the patients who showed Western blot-indeterminate results converted to HIV positive after follow-up. Thus, careful monitoring of patients with indeterminate Western blot results should be essential.
Abstract: The authors aimed to identify the correlation between quantitative hepatitis B surface antigen (HBsAg) and hepatitis B virus (HBV) DNA in Korean patients with a high level of HBsAg (>250 IU/ml) and characteristics of patients for whom quantitative HBsAg can be more suitably used to monitor HBV infection.
Abstract: Background: This study was performed to investigate on the genotypic frequencies
of p53 Arg72Pro polymorphism and the prevalence of p53 codon 249 mutation in
hepatocellular carcinoma patients.
Methods: Plasma DNAs were extracted from the samples of 44 early HCC cases,
24 chronic B-viral hepatitis patients and 27 healthy individuals. Serum levels of
AFP, PIVKA-II, and HBV DNA-positive rates among the study groups were also
compared. PCR-based restriction fragment length polymorphism method was used
to determine p53 Arg72Pro genotype and to detect codon 249 mutation.
Results: Serum AFP and PIVKA-II level, Edmondson grade, tumor size and
frequency of HBV DNA-positivity among HCC group according to Arg72Pro
genotypes showed no statistically significant difference. The frequencies of
Arg72Pro genotypes (Arg/Arg, Arg/Pro, Pro/Pro) were respectively as follows:
34.1%, 47.7%, 18.2% in HCC group; 29.2%, 54.2%, 16.7% in hepatitis group;
29.6%, 55.6%, 14.8% in control group. Pro homozygote genotype had a higher
risk for developing HCC by adjusted OR (1.529, 95% CI 0.325-7.193), but not
statistically significant (P=0.591). No codon 249 mutation was found among 44
HCC cases.
Conclusions: Pro homozygote was around 16% in all study groups, and did not
statistically increase risks to developing HCC. We suggest that Arg72Pro
polymorphism of p53 gene is not a significant risk factor in early
hepatocarcinogenesis.
Abstract: The anti-cyclic citrullinated peptide (anti-CCP) antibody has been increasingly used in the field of rheumatology, and various manufacturers have developed a variety of anti-CCP assays using mainly ELISA techniques. This study evaluated the performance of recently marketed automated chemiluminescence enzyme immunoassays for anti-CCP.
Abstract: Purpose: Early identification of causative agents in lower respiratory infection of pediatric patients can reduce morbidity and prevent an overuse of antimicrobials. Two kinds of multiplex polymerase chain reaction (PCR) and a commercial shell vial viral culture were performed to identify causative agents in pediatric patients. Materials and Methods: Nasopharyngeal aspirates of 220 children diagnosed with viral pneumonia were obtained. Two kinds of multiplex PCR (Seeplex(TM) RV detection kit, and Labopass(TM) RV detection kit), and a shell vial culture by R-Mix were performed. Results: Positive samples from 220 total samples by two multiplex PCRs were 52.7% and 46.4%, respectively. We also cultured 103 samples that showed positive results of the adenovirus, influenza virus, parainfluenza virus, and respiratory syncytial virus (RSV) by two multiplex PCR. The RSV was most frequently detected in 53.0% (Seeplex) and 51.7% (Labopass) of patients. The detection rate of adenovirus (AdV) was 10.3% and 12.1%, influenza virus (IFV) A and B was 12.5% and 3.4%, and parainfluenza virus (PIFV) 1, 2, and 3 were 2.9% and 2.6%. Shell vial cultures showed concordant results with each multiplex PCR by 96.1% and 77.7%, respectively. Sequencing results were 90% consistent with multiplex PCR. Conclusion: Multiplex PCR showed more positivity than the shell vial culture and it can be an effective primary test. Other complementary efforts such as viral cultures and sequencing analysis could be considered, according to clinical and laboratory conditions.
Abstract: We retrospectively analyzed the performance of the Architect HIV antigen/antibody (Ag/Ab) combination assay in a tertiary health care center with a situation of low HIV prevalence. The specificity and positive predictive value (PPV) were 99.78% and 31.21%, respectively. However, the specificity and PPV could increase to 99.99% and 89.70% using an arbitrary cutoff value.
Abstract: An automated hepatitis C virus (HCV) antigen (Ag) assay was evaluated with clinical samples. Determination of HCV Ag and RNA levels in 282 subjects using Abbott HCV Ag and Roche Cobas TaqMan assays revealed that these two tests were highly correlated (r = 0.9464). Thus, the HCV Ag assay could be an alternative test to quantitative reverse transcription-PCR.
Abstract: PURPOSE: The extraction of nucleic acid is initially a limiting step for successful molecular-based diagnostic workup. This study aims to compare the effectiveness of three automated DNA extraction systems for clinical laboratory use. MATERIALS AND METHODS: Venous blood samples from 22 healthy volunteers were analyzed using QIAamp Blood Mini Kit (Qiagen), MagNA Pure LC Nucleic Acid Isolation Kit I (Roche), and Magtration-Magnazorb DNA common kit-200N (PSS). The concentration of extracted DNAs was measured by NanoDrop ND-1000 (PeqLab). Also, extracted DNAs were confirmed by applying in direct agarose gel electrophoresis and were amplified by polymerase chain reaction (PCR) for human beta-globin gene. RESULTS: The corrected concentrations of extracted DNAs were 25.42 +/- 8.82 ng/microL (13.49-52.85 ng/microL) by QIAamp Blood Mini Kit (Qiagen), and 22.65 +/- 14.49 ng/microL (19.18-93.39 ng/microL) by MagNA Pure LC Nucleic Acid Isolation Kit I, and 22.35 +/- 6.47 ng/microL (12.57-35.08 ng/microL) by Magtration-Magnazorb DNA common kit-200N (PSS). No statistically significant difference was noticed among the three commercial kits (p > 0.05). Only the mean value of DNA purity through PSS was slightly lower than others. All the extracted DNAs were successfully identified in direct agarose gel electrophoresis. And all the product of beta-globin gene PCR showed a reproducible pattern of bands. CONCLUSION: The effectiveness of the three automated extraction systems is of an equivalent level and good enough to produce reasonable results. Each laboratory could select the automated system according to its clinical and laboratory conditions.
Abstract: To identify a desirable serum marker for screening tools for gastric cancer, we evaluated the validity of 3 biomarkers, namely, carcinoembryonic antigen (CEA), pepsinogens (PGs), and high sensitive C-reactive protein (hsCRP).
Abstract: We report the case of a 38-year-old man with acute promyelocytic leukemia (APL) showing a distinct breakpoint cluster region 2 (bcr2) variant transcript. Findings from bone marrow, cytogenetic, fluorescence in situ hybridization, and qualitative reverse transcriptase-polymerase chain reaction (RT-PCR) analyses were consistent with the diagnosis of APL. Although PCR products of size 841 bp and 984 bp were amplified from bone marrow specimen, the quantitative PCR (RQ-PCR) findings were negative. Given the discrepancy in PCR results, sequencing of PCR products was performed to determine the detailed composition of these fusion transcripts. By cloning and sequencing, we discovered that these two bands were isoforms, in which one exon (exon 5, 144 bp) of the PML gene was spliced out of the smaller products (minor PCR products); one sequence (G) insertion and one base substitution (T-->C) of PML exon 4 generate a stop codon in the smaller fusion transcript. In addition, a search of the Ensembl database revealed that these variant PML/RARA fusion transcripts were composed of exon 7a insertion of the PML gene and partial deletion (46 bp) of exon 3 of the RARA gene, in addition to inserted sequences of intron 7 of PML and genomic sequence ATCT of unknown origin at the fusion junction site. Although the biological significance of most atypical transcripts remains unclear, sequence analysis of these atypical products should be performed, to reveal the composition of such a fusion transcript and elucidate the molecular mechanism.
Abstract: We report on a 20-year-old man with acute myeloid leukemia (AML) showing a distinct novel CBFB/MYH11 variant fusion transcript. Initial results of bone marrow, chromosome, and flow cytometric analyses were not in accordance with the diagnosis of acute myelomonocytic leukemia with eosinophilia (AML-M4Eo) or AML with a CBFB/MYH11 rearrangement. However, results from 2 commercially available multiplex reverse transcriptase-polymerase chain reaction (RT-PCR) tests repeatedly showed an unusual PCR product from his bone marrow specimen. Not only does this case show a partial insertion of exon 6 of the CBFB (ENSG00000067955) gene, but it also involves novel breakpoints within both exon 6 of the CBFB gene and exon 28 (previously exon 7) of the MYH11 (ENSG00000133392) gene, which is regarded as a previously non-reported, new type (K-type) of CBFB/MYH11 fusion transcript. In addition, our study result was in agreement with the recent report of Schnittger et al. that rare fusion transcripts of CBFB/MYH11 are correlated with an atypical cytomorphology and other aberrant characteristics. Therefore, multiplex RT-PCR and sequence analysis of these atypical products should be performed to diagnose atypical AML with CBFB/MYH11 rearrangement, to predict prognosis of these patients as well as to elucidate the molecular mechanism.
Abstract: To reduce morbidity and mortality through integrated case management, a pilot study to detect respiratory viruses in patients with acute lower respiratory infections (ALRIs) was designed as part of a nationwide surveillance for this disease in Korea. The study population consisted of hospitalized patients under the age of 5 years with bronchiolitis, pneumonia, croup, or acute respiratory distress syndrome. A prospective 6-month study was performed. Two hundred and ninety-seven nasopharyngeal secretions were collected and multiplex reverse transcriptase polymerase chain reactions (RT-PCR)/polymerase chain reactions (PCR) were performed to detect respiratory viruses. If there were any positive RT-PCR/PCR results, viral cultures were proceeded for confirmation. Respiratory viruses were identified in 49.6% of 296 patients. The detection rates were as follows: respiratory syncytial virus (RSV) was the most commonly detected in 52.7% (87/165), human metapneumovirus (hMPV) in 15.8%, human corona virus (hCoV) in 5.5%, adenovirus in 9.7%, human bocavirus (hBoV) in 5.5%, parainfluenza virus (PIV) in 3.6%, rhinovirus (RV) in 4.2%, and the influenza virus in 3% of the patients with ALRIs. The consistent rate of positive results between RT-PCR and viral culture was 92% (105/114). Using our methods to detect viral causes seemed to be acceptable for the national surveillance of severe acute respiratory infections in infants and children.
Abstract: We describe a novel case of simultaneous karyotypic abnormalities of isochromosome 17q and trisomy 14 in a patient with myelodysplastic syndrome (MDS) in leukemic transformation. A 66-yr-old Korean man was admitted to Severance Hospital for evaluation of pancytopenia. On the basis of bone marrow studies at 3 different stages, he was diagnosed with MDS in leukemic transformation. Chromosome karyotyping repeatedly showed the same main clonal abnormalities, including isochromosome 17q and trisomy 14. Isochromosome 17q and trisomy 14 have each been reported as rare, nonrandom recurrent chromosomal abnormalities in patients with MDS showing a poor prognosis. To our knowledge, this is the first report of concurrent i(17)(q10) and trisomy 14 in a patient with MDS in leukemic transformation.
Abstract: We report a rare case of acute myeloid leukemia (AML) with t(6;11)(q15;q23) in a 50-year-old female showing a poor prognosis. Bone marrow biopsy revealed markedly hypercellular marrow with infiltrates of myeloblasts, consistent with AML-M2 morphology. The karyotype of this patient was 46,XX,t(6;11)(q15;q23) in all analyzed cells, and the results of fluorescence in situ hybridization (FISH) and multi-color FISH analysis confirmed this unique MLL rearrangement as a sole abnormality. To our knowledge, t(6;11)(q13 approximately q15;q23) is the most rare type of MLL rearrangement involving the long arm of chromosome 6. Only two cases with t(6;11)(q13;q23) and three cases with t(6;11)(q15;q23) have been reported, but detailed clinical or laboratory data were not available. From this report, it is apparent that in a cytogenetic laboratory, the accurate detection of a rare type of MLL rearrangement is very important in the differential diagnosis, prompt treatment, and prediction of prognosis of leukemias.
Abstract: To evaluate the usefulness of cytokine levels of peripheral blood in diabetic retinopathy (DR), demographic and biochemical parameters including low-density lipoprotein (LDL) diameter as well as cytokine profiles were analyzed in 74 patients with type 2 diabetes mellitus (DM), with DR (n=46) or without DR (n=28). DM duration was longer in the patients with DR than without (p<0.001). Serum glucose (p=0.005) and total cholesterol (p=0.029) levels were higher in DM patients with DR than DM patients without DR. Plasma LDL diameter, interleukin-6 (IL-6), and interleukin-8 (IL-8) showed significant differences among the different degrees of DR severity in analysis of variance (ANOVA) with no definite trend. The risk of DR in DM patients was decreased by an increase of interleukin-10 (IL-10) level [odds ratio (OR)=0.152; confidence interval (CI): 0.028-0.817]. Plasma LDL diameter was smaller and IL-6 and tumor necrosis factor-alpha (TNF-alpha) levels were higher in DM patients with proliferative diabetic retinopathy (PDR) compared to those with non-proliferative diabetic retinopathy (NPDR) (p<0.05). We found that higher IL-10 levels were related to lower risk of DR in DM patients. Levels of IL-6 and TNF-alpha as well as LDL diameter may be helpful in the prediction of PDR in DM patients with DR.
Abstract: BACKGROUND: Methicillin-resistant Staphylococcus aureus (MRSA) and some gram-negative bacilli are very prevalent nosocomial pathogens, commonly causing mixed infections, and are often resistant to multiple drugs. Arbekacin is an aminoglycoside used for the treatment of MRSA infections, but is also active against some gram-negative bacilli. The aim of this study was to determine in vitro activity of arbekacin against recent clinical isolates of staphylococci and gram-negative bacilli. Materials and METHODS: The strains were isolated from clinical specimens of patients at Severance Hospital in 2003. Antimicrobial susceptibility was tested by the Clinical and Laboratory Standards Institute agar dilution method. The following arbekacin breakpoints were used: susceptible, </=4 microg/mL; and resistant, >/=16 microg/mL . RESULTS: All isolates of staphylococci tested were inhibited by </=4 microg/mL of arbekacin, regardless of their methicillin susceptibility. The MIC90s of arbekacin, 1-4 microg/mL, were 8->32-fold and >32-128-fold lower than those of amikacin and gentamicin, respectively. The resistance rates of MRSA, methicillin-susceptible S. aureus, methicillin-resistant coagulase-negative staphylococci (CNS) and methicillin-susceptible CNS were 0% to arbekacin, 0-54% to amikacin, and 24-79% to gentamicin. The MIC90s of arbekacin for Escherichia coli and Citrobacter freundii, 1 microg/mL and 16 microg/mL, were 2-4-fold and 8-16-fold lower than those of amikacin and gentamicin, respectively. However, The MIC90s of arbekacin for other species of gram-negative bacilli, 64->128 microg/mL, were similar to those of other aminoglycosides. CONCLUSIONS: Arbekacin may be a useful alternative to glycopeptides for the treatment of monomicrobial methicillin-resistant staphylococcal infections, as well as mixed infections with gram-negative bacilli, as most isolates of E. coli, C. freundii and some other gram-negative bacilli were also susceptible to arbekacin.
