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.

Enzyme-Linked Immunosorbent Assay ; Hepatitis B Surface Antigens ; blood ; Humans ; Immunoassay ; methods ; Luminescent Measurements ; methods

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.

Blotting, Western ; methods ; China ; Enzyme-Linked Immunosorbent Assay ; methods ; HIV Infections ; diagnosis ; Humans ; Sensitivity and Specificity

In the present study, quids from La Cueva de los Muertos Chiquitos (CMC) were subjected to ELISA tests for 2 protozoan parasites, Toxoplasma gondii (n=45) and Trypanosoma cruzi (n=43). The people who occupied CMC, the Loma San Gabriel, lived throughout much of present-day Durango and Zacatecas in Mexico. The known pathoecology of these people puts them into at-risk categories for the transmission of T. gondii and T. cruzi. Human antibodies created in response to these 2 parasites can be detected in modern saliva using ELISA kits intended for use with human serum. For these reasons, quids were reconstituted and subjected to ELISA testing. All test wells yielded negative results. These results could be a factor of improper methods because there is no precedence for this work in the existing literature. The results could equally be a simple matter of parasite absence among those people who occupied CMC. A final consideration is the taphonomy of human antibodies and whether or not ELISA is a sufficient method for recovering antibodies from archaeological contexts. An additional ELISA test targeting secretory IgA (sIgA) was conducted to further examine the failure to detect parasite-induced antibodies from quids. Herein, the methods used for quid preparation and ELISA procedures are described so that they can be further developed by future researchers. The results are discussed in light of the potential future of quid analysis.
Antibodies ; Enzyme-Linked Immunosorbent Assay ; Humans ; Immunoglobulin A, Secretory ; Loma ; Methods ; Mexico ; Parasites ; Saliva ; Toxoplasma ; Trypanosoma cruzi

OBJECTIVESTo develop and evaluate a method for detection of the total antibodies against hemorrhagic fever with renal syndrome (HFRS) virus with improved sensitivity and simplified operation procedure.

METHODSThe nucleic proteins of hantavirus were used as coating antigens as well as detection antigens labeled with horse radish peroxidase (HRP). The operation protocol was established, optimized and compared with indirect fluorescence assay (IFA).

RESULTSThe specificity of this method was 100 percent in the test of different human sera and 4-8 times more sensitive than IFA. And, it is simpler without requiring any change of the reagents, different sources of samples did not affect the results of the test.

CONCLUSIONThis method is specific, sensitive and simple for detection of the total antibodies in sera against hantavirus, could be used for the screening of Hantavirus infection in human and host rodent animals.

Antibodies, Viral ; blood ; Enzyme-Linked Immunosorbent Assay ; methods ; Hantaan virus ; immunology ; Humans ; Reproducibility of Results ; Sensitivity and Specificity

IgG4-related disease (IgG4-RD) is a novel and rare autoimmune disease entity. Elevated serum IgG4 level is strongly suggestive of IgG4-RD. But it is still unknown whether serum IgG4 elevation commonly occurs in other autoimmune diseases. In this study, the serum IgG4 levels were detected by an established enzyme-linked immunosorbent assay (ELISA) in a variety of autoimmune diseases including systemic lupus erythematosus (SLE), Sjogren's syndrome (SS), polymyositis or dermatomyositis (PM/DM) and IgG4-RD. To evaluate the reliability of this ELISA system, some of our samples were sent to a lab in Kanazawa Medical University, Japan, and detected by using the nephelometric assay. The results showed that our findings were consistent with theirs. Moreover, it was found that the serum IgG4 levels were 0.23±0.16 g/L in 53 healthy controls, 0.16±0.15 g/L in 103 SLE patients, 0.22±0.18 g/L in 41 SS patients and 0.40±0.32 g/L in 21 PM/DM patients. No significant difference in the serum IgG4 level was observed among these groups (P>0.05). The serum IgG4 levels of two cases of IgG4-RD were 1.63 and 4.65 g/L respectively, and both decreased markedly after treatment with glucocorticoids. These data indicated that this established ELISA system can be used for detecting serum IgG4 levels. Elevated serum IgG4 levels help diagnose IgG4-RD and evaluate the curative effect of this condition rather than other autoimmune diseases.
Autoimmune Diseases ; blood ; diagnosis ; immunology ; Enzyme-Linked Immunosorbent Assay ; methods ; Humans ; Immunoglobulin G ; blood

OBJECTIVETo prepare the antibodies against salbutamol (SAL) with high sensitivity and to develop an indirect competitive enzyme-linked immunoassay (ic-ELISA) for fast detection of SAL.

METHODSThe New Zealand white rabbits were immunized with SAL in a small dose and long period mode. The method of ic-ELISA was optimized and adopted for the detection of a series of SAL samples, then the standard curve of SAL was established. The precision and the recoveries of the method were determined.

RESULTSThe antibodies with high sensitivity towards SAL were prepared with a IC50 of 12.21 ng/ml. The ic-ELISA method for SAL measurement was established, the recoveries of measurement was between 95%-105% and the CV was <3%.

CONCLUSIONThe antibodies against salbutamol have been prepared and an indirect competitive enzyme-linked immunoassay for fast and specific detection of SAL has been developed.

Albuterol ; analysis ; immunology ; Animals ; Antibodies ; immunology ; Enzyme-Linked Immunosorbent Assay ; methods ; Male ; Rabbits

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: 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