In Trang An polyclinics in Ha Noi, with ELISA technique, 35 healthy people (27 female aged 20-38, 8 male aged 25-37) undergone an examination of serum CA153-3 and 31 other healthy people (17 female aged 21-38, 14 male aged 22-41) an examination of resum CA125. The technique is complicated needing precaution and accuracy; for each examination, it is recommended a standard graphic with a standard sample determined antigen concentration. CA15-3 normal concentration is 2.37-18.17 U/ml, and CA125 is 7.17-38.37 U/ml
Enzyme-Linked Immunosorbent Assay ; methods ; serum

OBJECTIVETo evaluate chemiluminescent immunoassay (CLIA), electrochemiluminescent immunoassay (ECLIA) and ELISA in determination of low level HBsAg.

METHODSAccording to the standard of CLIA Architect i2000, 70 samples were divided into three groups by HBsAg concentration : <1 ng/ml, 1-5 ng/ml and >4 ng/ml. The samples were also determined by ECLIA MODULAR170 and ELISA, and the results were compared with those measured by CLIA Architect i2000.

RESULTSThe concordance rates of ECLIA MODULAR170 with Architect i2000 was 79.2% for <1 ng/ml group, 100% for 1-5 ng/ml group, 100% for >4 ng/ml group. And the concordance rates of ELISA with Architect i2000 was 0 % for <1 ng/ml group, 45.5% for 1-5 ng/ml group and 100 % for >4 ng/ml group.

CONCLUSIONFor determination of low level HBsAg,CLIA Architect i2000 and ECLIA MODULAR170 have high credibility. ELISA is also credible when HBsAg >5 ng/ml, but not for low level HBsAg, especially for HBsAg <1 ng/ml.

OBJECTIVETo identify a cost-efficient alternative antibody testing strategy for screening and confirmation of HIV infection by rapid simple tests (RSTs) and enzyme-linked immunosorbent assays (ELISAs).

METHODSFour RSTs (RST1, RST2, RST3, and RST4) and five ELISAs (ELISA1, ELISA2, ELISA3, ELISA4, and ELISA5) were evaluated in two phases by using banked and serum specimens prospectively collected at regional hospitals and voluntary counseling and testing (VCT) centers in Beijing. A total of 200 banked serum specimens were included in the first phase, including 62 HIV-positive, 127 HIV-negative and 11 indeterminate specimens. All specimens were tested by four RSTs and five ELISAs respectively. The second phase involved prospective testing of 389 routine specimens, including 92 HIV-positive, 287 HIV-negative, and 10 indeterminate specimens. All the specimens were tested by two RSTs (RST2 and RST4) and three ELISAs (ELISA1, ELISA3, and ELISA4), which were selected for their respective excellent sensitivity and/or specificity. Western blot (WB) was used as a gold standard for confirming the reactivity of all the specimens.

RESULTSSensitivity, specificity, and efficacy were calculated for each assay in two phases. In the first phase, four assays (ELISA4, RST2, RST3, and RST4) had a specificity of 100%. For the determination of efficacy, ELISA4, RST2, and RST4 were selected in the second phase. ELISA1 and ELISA3 which have a sensitivity of 95.9% and 93.2% respectively also entered this phase. In the second phase, all the five assays (ELISA1, ELISA3, ELISA4, RST2, and RST4) had a sensitivity and specifity of over 90%. ELISA1 had a sensitivity of 99% and ELISA4 a specificity of 99%.

CONCLUSIONThe sensitivity ELISA1 and the specificit of ELISA4 are comparable to ELISA/WB standard strategy. Application of this alternative testing strategy provides a cost-effective method for determining HIV prevalence in Beijing.

OBJECTIVE: α-synuclein, Nogo-A and Ubiquitin C-terminal hydrolase L1 (UCH-L1) have neuromodulatory roles for human brain. Therefore, abnormalities of these molecules are associated with neuropsychiatric disorders. Although some serum studies in the other disorders have been made, serum study of α-synuclein, Nogo-A and UCH-L1 is not present in patients with schizophrenia and healthy controls. Therefore, our aim was to compare serum levels of α-synuclein, Nogo-A and UCH-L1 of the patients with schizophrenia and healthy controls. METHODS: Forty-four patients with schizophrenia who is followed by psychotic disorders unit, and 40 healthy control were included in this study. Socio-demographic form and Positive and Negative Syndrome Scale (PANSS) was applied to patients, and sociodemographic form was applied to control group. Fasting bloods were collected and the serum levels of α-synuclein, Nogo-A and UCH-L1 were measured by ELISA method. RESULTS: Serum α-synuclein [patient: 12.73 (5.18–31.84) ng/mL; control: 41.77 (15.12–66.98) ng/mL], Nogo-A [patient: 33.58 (3.09–77.26) ng/mL; control: 286.05 (136.56–346.82) ng/mL] and UCH-L1 [patient: 5.26 (1.64–10.87) ng/mL; control: 20.48 (11.01–20.81) ng/mL] levels of the patients with schizophrenia were significianly lower than healthy controls (p<0.001). CONCLUSION: Our study results added new evidence for explaining the etiopathogenesis of schizophrenia on the basis of neurochemical markers.
Biomarkers ; Brain ; Enzyme-Linked Immunosorbent Assay ; Fasting ; Humans ; Methods ; Psychotic Disorders ; Schizophrenia* ; Ubiquitin Thiolesterase

PURPOSE: Homeless patients usually live outside and are therefore frequently exposed to injury and tetanus infection. Thus, after visits to an emergency department (ED) due to injury, homeless patients need to be vaccinated for the prevention of tetanus infection with tetanus immunoglobulin regardless of tetanus antibody titer or previous vaccination history. Because the exact history of previous tetanus vaccination in homeless patients is unclear, the tetanus antibody titer between homeless patients and general patients was assessed. METHODS: Subjects who visited the ED after injury from October 2008 to February 2010 were enrolled. All participants answered questions on age, gender, previous vaccination or prophylaxis history, and military service. The Tetanus Immunoglobulin G ELISA (Enzyme-linked immunosorbent assay) method was used for the analysis of serum samples. Propensity score-matched analysis was used to control for age, gender, previous vaccination or prophylaxis history, and military service. RESULTS: A total of 1325 samples were analyzed. There was 83 samples from homeless patients and 1242 samples from general patients. After matched analysis using the propensity score, 56 subjects were matched. The geometric mean titer of tetanus antibody was 0.204+/-0.392 IU/mL in homeless patients and 0.105+/-0.143 IU/mL in general patients (p=0.078). The proportion of patients with a safe tetanus antibody titer was 66.1 percent of homeless patients and 23.2 percent of general patients (p<0.001). CONCLUSION: Homeless patients had a higher mean titer and a statistically higher proportion had a safe titer compared to general patients.
Emergencies ; Enzyme-Linked Immunosorbent Assay ; Humans ; Immunoglobulin G ; Immunoglobulins ; Methods ; Military Personnel ; Propensity Score ; Tetanus* ; Vaccination

PURPOSE: Homeless patients usually live outside and are therefore frequently exposed to injury and tetanus infection. Thus, after visits to an emergency department (ED) due to injury, homeless patients need to be vaccinated for the prevention of tetanus infection with tetanus immunoglobulin regardless of tetanus antibody titer or previous vaccination history. Because the exact history of previous tetanus vaccination in homeless patients is unclear, the tetanus antibody titer between homeless patients and general patients was assessed. METHODS: Subjects who visited the ED after injury from October 2008 to February 2010 were enrolled. All participants answered questions on age, gender, previous vaccination or prophylaxis history, and military service. The Tetanus Immunoglobulin G ELISA (Enzyme-linked immunosorbent assay) method was used for the analysis of serum samples. Propensity score-matched analysis was used to control for age, gender, previous vaccination or prophylaxis history, and military service. RESULTS: A total of 1325 samples were analyzed. There was 83 samples from homeless patients and 1242 samples from general patients. After matched analysis using the propensity score, 56 subjects were matched. The geometric mean titer of tetanus antibody was 0.204+/-0.392 IU/mL in homeless patients and 0.105+/-0.143 IU/mL in general patients (p=0.078). The proportion of patients with a safe tetanus antibody titer was 66.1 percent of homeless patients and 23.2 percent of general patients (p<0.001). CONCLUSION: Homeless patients had a higher mean titer and a statistically higher proportion had a safe titer compared to general patients.
Emergencies ; Enzyme-Linked Immunosorbent Assay ; Humans ; Immunoglobulin G ; Immunoglobulins ; Methods ; Military Personnel ; Propensity Score ; Tetanus* ; Vaccination

PURPOSE: Tuberculosis (TB) is a major infectious disease and is responsible for two million deaths annually. For the identification and quantitation of Mycobacterium tuberculosis (M. tuberculosis), a causative agent of TB, a sandwich enzyme-linked immunosorbent assay (ELISA) against the MPT64 protein of M. tuberculosis, an antigen marker of the M. tuberculosis complex, was developed. MATERIALS AND METHODS: The MPT64 protein was expressed, and anti-MPT64 monoclonal antibodies were prepared. A sandwich ELISA was established using recombinant MPT64 protein and anti-MPT64 monoclonal antibodies. The sandwich MPT64 ELISA was evaluated using reference and clinical mycobacterial strains. RESULTS: The sandwich MPT64 ELISA detected MPT64 protein from 2.1 ng/mL to 250 ng/mL (equivalent to 1.7x10(4) CFU/mL and 2.0x10(6) CFU/mL). All 389 clinical M. tuberculosis isolates tested positive in the sandwich MPT64 ELISA (sensitivity, 100%), and the assay showed no cross reactivity to any tested nontuberculous mycobacterial strain (specificity, 100%). CONCLUSION: The sandwich MPT64 ELISA is a highly sensitive and quantitative test for MPT64 protein, which can identify M. tuberculosis.
Antigens, Bacterial/*analysis/immunology ; Enzyme-Linked Immunosorbent Assay/*methods ; Mycobacterium tuberculosis/*immunology

PURPOSE: To evaluate a recently marketed commercial glycoprotein enzyme-linked immunosorbent assay (gpEIA) kit, the VaccZyme(TM) VZV gpEIA, for measuring the immunity of varicella-vaccinated children. MATERIALS AND METHODS: We investigated the accuracy and reproducibility of the VaccZyme(TM) VZV gpEIA kit for the detection of antibodies to VZV. We also examined the sensitivity, specificity, and correlation between antibody titers calculated with gpEIA versus fluorescent antibody to membrane antigen (FAMA) by using sera of 349 children, ranging from 1 to 6 years old. RESULTS: VaccZyme(TM) VZV gpEIA gave precise and reproducible intra- and inter-assay results. FAMA and gpEIA titers showed a linear correlation (Pearson correlation coefficient=0.987). The sensitivity and specificity of the VaccZyme(TM) gpEIA was 31.4% and 100%, respectively, when the guidelines of the gpEIA (<100 mIU/mL) and FAMA 1:4 were adopted as cutoff values. However, the maximum sensitivity and specificity were 88.9% and 95.1%, respectively, with the highest correlation (kappa=0.840), if the cutoff values were set with gpEIA at 49.7 mIU/mL and FAMA 1:16. CONCLUSION: These results demonstrate that the VaccZyme(TM) VZV gpEIA kit gave precise and reproducible data for measuring antibody titer after varicella vaccination. The results also showed that the antibody titer calculated with the VaccZyme(TM) gpEIA kit strongly correlated with the FAMA titer. However, cutoff values should be re-optimized for the evaluation of vaccine immunity.
Antibodies ; Chickenpox* ; Child ; Enzyme-Linked Immunosorbent Assay* ; Glycoproteins* ; Humans ; Membranes ; Methods ; Sensitivity and Specificity ; Vaccination

BACKGROUND: As stated in ‘The Action Strategy for Tuberculosis-Free Korea,’ last March, high-throughput, large-scale analytical instruments for interferon gamma release assays (IGRA) are demanded by many clinical laboratories using the QuantiFERON-TB Gold In-Tube assay (Cellestis/Qiagen, Australia). Agility (Dynex Technologies, USA) is an automated high-throughput enzyme linked immunosorbent assay analyser. The present study aimed to evaluate its accuracy and speed. METHODS: Pooled plasma was prepared using samples obtained after IGRA testing. Analyses of precision, linearity, cut-off evaluation, and comparison with conventional methods were performed for multiple Agility instruments according to the Clinical and Laboratory Standards Institute EP5-A3, EP6-A, EP9-A3 and EP12-A2 guidelines. The turnaround time and throughput were also analysed. RESULTS: The coefficient of variation range was 2.48%–4.0%, 7.01%–11.17%, and 9.69%–14.84% for the repeatability, between-run precision, and between-day precision analyses, respectively. The linearity ranged from 0 to 10.541. Comparison analysis presented a high concordance of Agility with the conventional instrument, DS2 (Dynex Technologies), and manual method for IGRA. The cut-off value of 0.35 IU/mL was well compatible with the C50. It was identified that the C50±20% contained the C5–C95 interval. The average turnaround time was 3.84 hours, from the submission of pre-treated samples to the reporting of results. The throughput was determined to be 290 tests during a routine working time of 8 hours. CONCLUSIONS: Agility showed high precision, linearity, concordance, and had a 2.5 times faster throughput than with the conventional and manual method. It could be useful for large-scale IGRA testing in latent tuberculosis infection screening project. Samples within C50±20% are suspected to show relatively low reporducible results of high inversion between postivie and negative.
Enzyme-Linked Immunosorbent Assay ; Interferon-gamma Release Tests ; Interferons ; Latent Tuberculosis ; Mass Screening ; Methods ; Plasma

A competitive enzyme-linked immunosorbent assay (ELISA) was developed to determine ruscogenin (RUS) by using the monoclonal antibody (McAb). The monoclonal antibody against RUS, secreted from the established hybridoma cell lines, was identified as being of the IgG1 isotype. The McAb exhibited high specificity to RUS, showing a very slight cross reactivity with diosgenin (15.7%), and no cross-reactivity to sarsasapogenin, diammonium glycyrrhizinate, oleanolic acid and notoginsenoside R1. The established ELISA, at an IC50 value of 157.55 ng·mL(-1) and a detection limit (IC20) of 20.57 ng·mL(-1), was compared with HPLC analyses, and a good correlation between ELISA and HPLC-ELSD analyses of RUS in the extract of Radix Ophiopogonis was obtained. The experimental data indicated that the ELISA method exhibits more advantages over HPLC-ELSD, such as low detection limit, high specificity, low background, and no requirement for sample pre-treatment, and is more suitable for the determination of natural components in Chinese traditional medicines and in biological samples for pharmacokinetic studies.
Antibodies, Monoclonal ; analysis ; Drugs, Chinese Herbal ; analysis ; Enzyme-Linked Immunosorbent Assay ; methods ; Sensitivity and Specificity ; Spirostans ; analysis