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 5 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.

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.Since the first HIV test was introduced in 1985, the performances of HIV screening assays have improved continuously. In particular, the HIV antigen/antibody (Ag/Ab) combination assays launched in 1997 have shortened the window period by 4 to 5 days compared to those of the previous antibody-alone enzyme immunoassays (8). After the improvement of the detection limit of the p24 antigen enzyme immunoassay (EIA) to equivalent to that of the single-antigen EIA, HIV Ag/Ab combination assays have been implemented in numerous laboratories throughout the world (7, 9).The Centers for Disease Control and Prevention (CDC) recommended the expansion of HIV antibody testing from targeted, risk-based testing to universal screening of all adults aged 13 to 64 years in health care settings (2). Although the revised recommendation could be effective in identifying the maximum possible number of HIV-infected people from a public health perspective, there are concerns about false-positive results (3, 11). False-positive HIV screening results could cause substantial psychological distress while waiting for a confirmatory test (10). Guinn estimated the positive predictive value (PPV) of rapid HIV testing in Oregon with 100% sensitivity and 99.9% specificity to be 29% (3). The rate of falsepositive results could be dramatically increased in situations of extremely low HIV prevalence. Shima-Sano et al. reported that the PPV of HIV screening results in pregnant women is only 3.7% (5).According to a report by the Korea Centers for Disease Control and Prevention, 6,120 individuals have been diagnosed with an HIV infection between 1985 and 2008 in Korea (4). Although the number of newly diagnosed HIV infections has increased, the cumulative number of HIV-infected individuals and prevalence were lower than those in other countries. There has been limited study on the rate of false-positive results in HIV screening tests using an automated HIV Ag/Ab combination assay. In the present study, we retrospectively analyzed the performance of an automated HIV Ag/Ab combination assay in a tertiary health care center with a situation of low HIV prevalence.

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-B 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-B activity. Other regulatory pathways seem to exist for NF-B activation. In conclusion, a specific HBx mutation may contribute to HCC development by activating NF-B 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.Hepatitis B virus (HBV) infection is a global health issue, with two billion people infected worldwide and 350 million suffering from chronic HBV infection (18). Chronic HBV infection is known as the most common underlying etiology of hepatocellular carcinoma (HCC) (2) and can further progress to liver cirrhosis (LC) and HCC. However, the pathogenesis of these HBV-related diseases has not been fully clarified. The disease progression may depend on complex and multistep mechanisms.In an HBV management consensus report, the National Institutes of Health (NIH) suggested risk factors associated with the increased risk of HCC and cirrhosis in chronic HBVinfected patients (20). That report described the HCC risk based on demographic factors, environmental factors, and viral factors and classified them using the scoring system of evidence-based studies. The association between HBV mutations and HCC has not been fully investigated. Recently, the HBV PreS mutations C1653T, T1753V, and A1762T/G1764A were reported as strong risk factors for HCC in a meta-analysis (19). Mutations of HBV have been considered as an escape mechanism from the host immune system and may affect the oncogenic potential of chronic HBV diseases.HBV X is the smallest of four kinds of HBV functional genes, but it expresses a 154-amino-acid multifunctional protein (HBx), with an N-terminal negative regulatory/antiapoptotic domain and a C-termin...

PurposeThe 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 MethodsVenous 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.ResultsThe corrected concentrations of extracted DNAs were 25.42 ± 8.82 ng/µL (13.49-52.85 ng/µL) by QIAamp® Blood Mini Kit (Qiagen), and 22.65 ± 14.49 ng/µL (19.18-93.39 ng/µL) by MagNA Pure LC Nucleic Acid Isolation Kit I, and 22.35 ± 6.47 ng/µL (12.57-35.08 ng/µL) 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.ConclusionThe 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.

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