Immunological studies of ribonuclease C from human urine.

1981 ◽  
Vol 27 (8) ◽  
pp. 1362-1367 ◽  
Author(s):  
J W Cranston ◽  
H C Hoover ◽  
E R Crisp
Keyword(s):  
Pancreatic Carcinoma ◽  
Human Urine ◽  
Rnase A ◽  
Serum Samples ◽  
Positive Correlation ◽  
Alternative Method ◽  
Normal Individuals ◽  
Human Sera ◽  
Pancreatic Rnase

Abstract Antiserum to human urinary RNase C [ribonuclease (pancreatic), EC 3.1.27.5], developed in rabbits, was used to characterize this enzyme through studies of inhibition of RNase C-catalyzed poly(C) hydrolysis and of competition in a RIA. By either assay, the antiserum failed to cross react with human urinary RNase U (EC 3.1.27.-) or bovine pancreatic RNase A (EC 3.1.27.5). RNase C is immunologically identical to the poly(C)-active RNase in various human sera, including samples obtained from normal individuals, patients with pancreatic carcinoma, pancreatitis, or other malignant and nonmalignant diseases. This conclusion is based on the finding of superimposable antibody dose-inhibition curves for poly(C) hydrolysis and parallel competition RIA curves for RNase C and the various sera. There was a positive correlation (r = 0.73; p less than 0.001) between concentrations of RNase C as determined by poly(C) hydrolysis and competition RIA in serum samples from 102 patients. Therefore, the latter technique provides al alternative method for measuring RNase C in sera.

scholarly journals Antibodies to Highly Pathogenic A/H5Nx (Clade 2.3.4.4) Influenza Viruses in the Sera of Vietnamese Residents

Pathogens ◽  
2021 ◽  
Vol 10 (4) ◽  
pp. 394
Author(s):  
Tatyana Ilyicheva ◽  
Vasily Marchenko ◽  
Olga Pyankova ◽  
Anastasia Moiseeva ◽  
Tran Thi Nhai ◽  
...  
Keyword(s):  
Influenza Virus ◽  
Avian Influenza ◽  
Wide Spectrum ◽  
Influenza Viruses ◽  
Serum Samples ◽  
Socialist Republic ◽  
Highly Pathogenic ◽  
Virus Variant ◽  
Human Sera ◽  
Hemagglutinin Inhibition

To cause a pandemic, an influenza virus has to overcome two main barriers. First, the virus has to be antigenically new to humans. Second, the virus has to be directly transmitted from humans to humans. Thus, if the avian influenza virus is able to pass the second barrier, it could cause a pandemic, since there is no immunity to avian influenza in the human population. To determine whether the adaptation process is ongoing, analyses of human sera could be conducted in populations inhabiting regions where pandemic virus variant emergence is highly possible. This study aimed to analyze the sera of Vietnamese residents using hemagglutinin inhibition reaction (HI) and microneutralization (MN) with A/H5Nx (clade 2.3.4.4) influenza viruses isolated in Vietnam and the Russian Federation in 2017–2018. In this study, we used sera from 295 residents of the Socialist Republic of Vietnam collected from three groups: 52 samples were collected from households in Nam Dinh province, where poultry deaths have been reported (2017); 96 (2017) and 147 (2018) samples were collected from patients with somatic but not infectious diseases in Hanoi. In all, 65 serum samples were positive for HI, at least to one H5 virus used in the study. In MN, 47 serum samples neutralizing one or two viruses at dilutions of 1/40 or higher were identified. We postulate that the rapidly evolving A/H5Nx (clade 2.3.4.4) influenza virus is possibly gradually adapting to the human host, insofar as healthy individuals have antibodies to a wide spectrum of variants of that subtype.

Hepatitis C virus core antigen assay: an alternative method for hepatitis C diagnosis

2016 ◽  
Vol 54 (2) ◽  
pp. 279-285 ◽  
Author(s):  
Lijuan Wang ◽  
Hong Lv ◽  
Guojun Zhang
Keyword(s):  
Hepatitis C ◽  
Positive Predictive Value ◽  
Predictive Value ◽  
Characteristic Curve ◽  
High Specificity ◽  
Receiver Operator Characteristic Curve ◽  
Core Antigen ◽  
Hcv Rna ◽  
Serum Samples ◽  
Alternative Method

Background The study aimed to evaluate a fully automated chemiluminescent immunoassay and compared it with a quantitative RNA assay and anti-HCV assay to verify the utility of this automated Ag assay as an alternative method for hepatitis C diagnosis. Methods A total of 229 serum samples previously tested for anti-HCV concentrations by the Architect Anti-HCV assay, were selected for HCV RNA testing by real time RT-PCR kit (Shanghai ZJ Bio-Tec Co., Ltd) and 125 specimens were tested for HCV Ag by the Architect HCV core Antigen kit. Results The log10 HCVAg and HCV RNA concentrations were highly correlated [ r = 0.834); with HCV RNA as the comparator test, HCVAg had 100% specificity, 100% positive predictive value (PPV) and 94.8% sensitivity. We found 1 pg/mL of total HCV core Ag is equivalent to approximately 6607HCV RNA international units (IU)/mL. Receiver operator characteristic curve analysis showed that the area under the curve of HCV core Ag (0.989) was greater than HCV Ab (0.871). HCV Ag concentrations and RNA-to-Ag ratio of the groups for HCV RNA concentrations ≤105 and >105 IU/mL were both significantly different from each other ( P < 0.05). Conclusion The Architect HCV core Ag assay may be an alternative method for hepatitis C diagnosis, performed on the same analytical platform and sample as the anti-HCV assay, shortening the diagnostic window period, demonstrating good correlation with HCV RNA assay with high specificity and positive predictive value.

scholarly journals Standardization of dot-ELISA for the serological diagnosis of toxocariasis and comparision of the assay with ELISA

1992 ◽  
Vol 34 (1) ◽  
pp. 55-60 ◽  
Author(s):  
Eide Dias Camargo ◽  
Paulo Mutuko Nakamura ◽  
Adelaide José Vaz ◽  
Marcos Vinícius da Silva ◽  
Pedro Paulo Chieffi ◽  
...  
Keyword(s):  
Low Cost ◽  
Clinical Signs ◽  
Toxocara Canis ◽  
Enzyme Linked Immunosorbent Assay ◽  
Serum Samples ◽  
Specific Antibodies ◽  
Normal Individuals ◽  
Before And After ◽  
Dot Elisa ◽  
Stability Time

The dot-enzyme-linked immunosorbent assay (dot-ELISA) was standardized using somatic (S) and excretory-secretory (ES) antigens of Toxocara-canis for the detection of specific antibodies in 22 serum samples from children aged 1 to 15 years, with clinical signs of toxocariasis. Fourteen serum samples from apparently normal individuals and 28 sera from patients with other pathologies were used as controls. All samples were used before and after absorption with Ascaris suum extract. When the results were evaluated in comparison with ELISA, the two tests were found to have similar sensitivity, but dot-ELISA was found to be more specific in the presence of the two antigens studied. Dot-ELISA proved to be effective for the diagnosis of human toxocariasis, presenting advantages in terms of yield, stability, time and ease of execution and low cost.

scholarly journals Immunodiagnosis of human neurocysticercosis by using semi-purified scolex antigens from Taenia solium cysticerci

2007 ◽  
Vol 40 (2) ◽  
pp. 163-169 ◽  
Author(s):  
Francesco Iudici Neto ◽  
Geraldo Pianetti-Filho ◽  
Ricardo Nascimento Araújo ◽  
Evaldo Nascimento
Keyword(s):  
Sensitivity And Specificity ◽  
Excellent Agreement ◽  
Taenia Solium ◽  
Polyacrylamide Gel ◽  
Parasitic Diseases ◽  
Serum Samples ◽  
Serological Screening ◽  
Crude Antigen ◽  
Normal Individuals ◽  
Confirmatory Tests

Crude antigen and semi-purified proteins from scolices of Taenia solium cysticerci were evaluated for the immunodiagnosis of human neurocysticercosis neurocysticercosis. Semi-purified proteins obtained by electrophoresis on polyacrylamide gel and by electroelution were tested by means of the immunoenzymatic reaction against sera from normal individuals and from patients with neurocysticercosis or other parasitic diseases. The 100kDa protein provided 100% sensitivity and specificity in the immunodiagnosis. When 95 or 26kDa proteins were used, 95 and 100% sensitivity and specificity were obtained, respectively. The assays involving crude antigen and sera from normal individuals or from patients with neurocysticercosis, diluted to 1:256, gave excellent agreement with those in which 100, 95 or 26kDa proteins were tested against the same serum samples diluted to 1:64. (Kappa: 0.95 to 1.00). Crude scolex antigen may be useful for serological screening, while 100, 95 or 26kDa protein can be used in confirmatory tests on neurocysticercosis-positive cases.

scholarly journals Recombinant Dense Granular Protein (GRA5) for Detection of Human Toxoplasmosis by Western Blot

2014 ◽  
Vol 2014 ◽  
pp. 1-8 ◽  
Author(s):  
Xiao Teng Ching ◽  
Yee Ling Lau ◽  
Mun Yik Fong ◽  
Veeranoot Nissapatorn ◽  
Hemah Andiappan
Keyword(s):  
Western Blot ◽  
Affinity Purification ◽  
Expression System ◽  
Economic Loss ◽  
Serum Samples ◽  
Serological Tests ◽  
Human Patient ◽  
Prokaryotic Expression System ◽  
Human Sera ◽  
Lysate Antigens

Toxoplasma gondiiinfects all warm-blooded animals, including humans, causing serious public health problems and great economic loss for the food industry. Commonly used serological tests require costly and hazardous preparation of wholeToxoplasmalysate antigens from tachyzoites. Here, we have evaluated an alternative method for antigen production, which involved a prokaryotic expression system. Specifically, we expressedT. gondiidense granular protein-5 (GRA5) inEscherichia coliand isolated it by affinity purification. The serodiagnostic potential of the purified recombinant GRA5 (rGRA5) was tested through Western blot analysis against 212 human patient serum samples. We found that rGRA5 protein was 100% specific for analysis of toxoplasmosis-negative human sera. Also, rGRA5 was able to detect acute and chronicT. gondiiinfections (sensitivities of 46.8% and 61.2%, resp.).

scholarly journals Semiautomated microtiter plate assay for monitoring peptidylprolyl cis/trans isomerase activity in normal and pathological human sera

1998 ◽  
Vol 44 (3) ◽  
pp. 502-508 ◽  
Author(s):  
Gerhard Küllertz ◽  
Sabine Lüthe ◽  
Gunter Fischer
Keyword(s):  
Biological Fluids ◽  
Microtiter Plate ◽  
Serum Samples ◽  
Ppiase Activity ◽  
Microtiter Plate Assay ◽  
Isomerase Activity ◽  
Microtiter Plates ◽  
Activity Factor ◽  
Healthy Donors ◽  
Human Sera

Abstract An UV/VIS spectrophotometric assay technique was developed that was able to routinely monitor peptidylprolyl cis/trans isomerase (PPIase) activity of biological fluids in 96-well microtiter plates. The assay, based on monitoring the cis-to-trans isomerization of succinyl-Phe-cisPro-Phe-4-nitroanilide as substrate in a chymotrypsin-coupled reaction, yields a throughput of 96 samples per 30 min. The assay’s capacity was exemplified by dealing with the PPIase activity in several normal and pathological human sera. Reference values of 151 healthy subjects (83 females, 69 males, 17 to 60 years old) were found to possess significant sex-specific differences. PPIase activity factor K of the sera was significantly greater in males (5th, 50th, 95th percentiles: 17, 36, 55 K) than females (14, 30, 48 K). PPIase activities of sera from healthy donors (n = 151) were significantly higher (Mann–Whitney rank-sum test P &lt;0.0001) than those of patients (n = 47). PPIase activity in serum samples stored at 4 °C was stable for at least 20 h.

scholarly journals Antibody response in patients with Gaucher disease after repeated infusion with macrophage-targeted glucocerebrosidase

Blood ◽  
1993 ◽  
Vol 82 (5) ◽  
pp. 1402-1409 ◽  
Author(s):  
SM Richards ◽  
TA Olson ◽  
JM McPherson
Keyword(s):  
Gaucher Disease ◽  
Mean Value ◽  
Patient Treatment ◽  
Enzyme Linked Immunosorbent Assay ◽  
Serum Samples ◽  
Enzyme Replacement ◽  
Human Sera ◽  
Normal Human ◽  
Igg1 Subclass

Abstract Recent clinical data have shown that enzyme replacement therapy with macrophage-targeted glucocerebrosidase (GCR) can be effective in treating type 1 Gaucher disease. Sera from 262 patients, repeatedly infused with GCR, were assessed for the presence of antibodies to this therapeutic protein. Patient serum samples obtained at 3-month intervals were assessed by enzyme-linked immunosorbent assay and those with values greater than two standard deviations above the mean value obtained with a pool of normal human sera were further characterized by radioimmunoprecipitation. At the time of these analyses, the duration of patient treatment varied from 3 months to approximately 3 years. Of the 262 patients analyzed, 34 (12.9%) showed IgG antibodies, as confirmed by radioimmunoprecipitation. All patients who seroconverted did so within 1 year of treatment. The predominant antibody developed was the IgG1 subclass. Fourteen patients in the study experienced periodic symptoms suggestive of immediate hypersensitivity. Nine of these 14 patients had antibody to GCR as determined by radioimmunoprecipitation, whereas 5 patients were antibody negative. There was no evidence of the development of IgE antibodies in these 14 patients. The presence of GCR antibodies did not appear to effect efficacy of therapy in any of the patients treated to date.

Reference intervals and biological variation for kallikrein 6: influence of age and renal failure

2012 ◽  
Vol 50 (5) ◽  
Author(s):  
Eduardo Martínez-Morillo ◽  
Anastasia Diamandis ◽  
Eleftherios P. Diamandis
Keyword(s):  
Renal Failure ◽  
Significant Positive Correlation ◽  
Reference Interval ◽  
Serum Marker ◽  
Reference Intervals ◽  
Biological Variation ◽  
Serum Samples ◽  
Positive Correlation ◽  
Reference Change Value ◽  
Kallikrein 6

AbstractKallikrein 6 (KLK6) is a serine protease involved in numerous cellular processes, up-regulated in many cancers and associated with some neurodegenerative disorders. The aim of this study was to establish a reference interval and estimate the biological variation of KLK6 in serum samples of adults. Furthermore, levels of this protein in patients with renal failure were also studied.Serum samples from healthy volunteers (n=136) were collected. Between 15 and 18 additional samples from four of these subjects were obtained over a period of 2 months. Samples from individuals (n=1043) who visited the University Health Network for a routine check-up were collected to study the association between KLK6 with age and gender. Samples from patients with renal failure (n=106) were also obtained and KLK6 and creatinine concentrations were analyzed by ELISA and an automated enzymatic method, respectively.The reference interval was established to be 1.04–3.93 ng/mL. The index of individuality was 0.43 and the reference change value was 35%. Only two serum samples would be required to estimate the homeostatic setting point of an individual. There is a weak but highly significant positive correlation between KLK6 and age (p<0.0001). Furthermore, there is a significant positive correlation between serum concentrations of KLK6 and creatinine (p<0.0001), in patients with renal failure.The established reference interval for KLK6 and the estimation of its biological variation will further aid in the clinical use of this protein as a serum marker of malignancy and other diseases.

Isolation of 2-Acetamido-1-β-(L-β-Aspartamido)-1,2-Dideoxy-D-Glucose from Normal Human Urine

1972 ◽  
Vol 18 (9) ◽  
pp. 951-955 ◽  
Author(s):  
Barbara O’Neill Rowley ◽  
Paul B Hamilton
Keyword(s):  
Ion Exchange ◽  
Human Urine ◽  
Ion Exchange Chromatography ◽  
Exchange Chromatography ◽  
Normal Individuals ◽  
Normal Human ◽  
Elution Behavior ◽  
Solvent Systems ◽  
And Migration ◽  
Layer Chromatography

Abstract A glycopeptide was isolated from normal human urine by fractionation on a column of Sephadex G-10 and preparative ion-exchange chromatography. Elution behavior during ion-exchange chromatography in two different solvent systems, amino acids formed upon hydrolysis, and migration on high-voltage electrophoresis and thin-layer chromatography were essentially identical for this substance and for authentic 2-acetamido-l-β-(L-β-aspartamido)-1,2-dideoxy-D-glucose. A technique was developed to permit analytical-scale fractionation of individual urines followed by analysis for this glycopeptide; urine from two normal individuals contained 7 and 11 µmol of 2-acetamido-1-β-(L-β-aspartamido)-1,2-dideoxy-D-glucose per liter.

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