scholarly journals Evaluation of the Optilite® analyser for determination of total complement activity and C3 and C4 fractions

2019 ◽  
Vol 77 (4) ◽  
pp. 447-452
Author(s):  
Benoit Nespola ◽  
Hélène Comitogianni ◽  
Isabelle Jahn ◽  
Joëlle Goetz
Keyword(s):  
Complement Activity ◽  
Total Complement

Automated homogeneous liposome-based assay system for total complement activity

1995 ◽  
Vol 41 (4) ◽  
pp. 586-590 ◽  
Author(s):  
S Yamamoto ◽  
K Kubotsu ◽  
M Kida ◽  
K Kondo ◽  
S Matsuura ◽  
...  
Keyword(s):  
Serum Proteins ◽  
M Protein ◽  
Homogeneous Population ◽  
Complement Activity ◽  
Activity Test ◽  
Human Sera ◽  
Total Complement ◽  
Hemolytic Complement Activity ◽  
Glucose 6 Phosphate Dehydrogenase

Abstract We developed an automated homogeneous immunoassay, based on immune lysis of dinitrophenyl (DNP)-labeled liposomes, for measuring total complement activity. Liposome lysis caused by complement activity was detected spectrophotometrically from entrapped glucose-6-phosphate dehydrogenase activity. Complement activity in human sera was quantified by comparison with a calibration curve. For ease of application to fully automated routine clinical analyzers, we adopted a two-reagent system, one reagent containing a homogeneous population of small DNP-labeled liposomes and one containing antibody/substrate. This system required calibration only once a week. Within-run and between-run CVs were 0.4-1.3% (n = 10) and 1.8-4.7% (n = 10), respectively. Serum results were linear upon dilution (with saline) over a twofold range. Bilirubin, hemoglobin, Intrafat, and serum proteins such as rheumatoid factor, M protein, IgG, and IgA did not affect the assay results. The results (y) correlated well with those from a hemolytic complement activity test (x): y = 1.05x - 1.14, r = 0.92, on 66 samples in the range < 10- > 50 kU/L. This method should therefore be of great use for the determination of complement activity.

scholarly journals A Flow Cytometry-Based Assay for the Measurement of Total Complement Activity in the Serum and Clinical Practice

2022 ◽  
Vol 2022 ◽  
pp. 1-12
Author(s):  
Xuewei Ding ◽  
Shijun Li ◽  
Hui Liu
Keyword(s):  
Flow Cytometry ◽  
Nephrotic Syndrome ◽  
Linear Relationship ◽  
High Volume ◽  
Healthy Individuals ◽  
Serum Samples ◽  
Complement Activity ◽  
Significant Difference ◽  
Cell Groups ◽  
Total Complement

Objective. To develop a novel sensitive and accurate assay suitable for high-volume testing of the total complement activity in the serum for clinical laboratories. Methods. The total complement activity (TCA) to be measured was quantified by detecting the number of fragments produced by erythrocyte lysis and the erythrocyte fragmentation index (EFI), indicating TCA. EFI = M × M 2 / M 1 + M 2 , where M is the number of erythrocyte fragments (removed from the background), M 1 is the number of unagglutinated red cells, M 2 is the number of agglutinated red cell groups, and M 2 / M 1 + M 2 is the agglutination coefficient indicating the degree of erythrocyte agglutination. Mild changes in hemolysin and erythrocyte concentrations were made to optimize the testing conditions. The same serum samples were tested for 10 consecutive days to determine the stability of the experimental results. Serum EFI was detected in both nephrotic syndrome patients and healthy subjects. Results. There was a linear relationship between hemolysin and erythrocyte agglutination ( r = 0.999 , P < 0.001 ). A good linear relationship existed between EFI and TCA ( r = 0.991 , P < 0.001 ). The results were not affected by slight fluctuations in the concentrations of hemolysin or erythrocytes. The interbatch CV = 8.6 % of the test results showed good stability. There was a significant difference in the EFI between nephrotic syndrome patients and healthy individuals, P < 0.001 , and EFI was reduced in nephrotic syndrome patients compared to healthy individuals. Conclusion. The flow cytometry-based assay for TCA was sensitive and accurate and had potential value for clinical application.

Computer-Assisted Kinetic Assay for Quantification of Total Complement Activity

1991 ◽  
Vol 8 (2) ◽  
pp. 92-103 ◽  
Author(s):  
M. Abbal ◽  
J. Tkaczuk ◽  
C. Praud ◽  
F. Msayeh ◽  
E. Ohayon
Keyword(s):  
Computer Assisted ◽  
Kinetic Assay ◽  
Complement Activity ◽  
Total Complement

scholarly journals THE DEVELOPMENT OF TOTAL COMPLEMENT ACTIVITY ASSAY BASED ON COMPLEMENT-DEPENDENT IMMUNE LIPOSOME LYSIS

2015 ◽  
Vol 69 (3-4) ◽  
pp. 24-30
Author(s):  
S. N. Skopinskaya ◽  
S. P. Yarkov ◽  
E. N. Khramov ◽  
A. V. Antashev
Keyword(s):  
Blood Serum ◽  
Activity Assay ◽  
Animal Blood ◽  
Complement Activity ◽  
Sheep Erythrocytes ◽  
Wide Range ◽  
Total Complement ◽  
Sulforhodamine 101 ◽  
Internal Volume ◽  
Definition Of

Background: The purpose of work was development of a fast and reproduced procedure for measurement of the total complement activity (ТСА) in human or animal blood serum. Materials and methods: Steady at storage liposomes preparations, which surface sensitized 2,4-DNP haptens, and the internal volume contains calceine or sulforhodamine 101 are obtained. Complement-dependent immune lysis of liposomes at presence of the anti-2,4-DNP immunoglobulines and complement preparations from animals are investigated. Results: It is shown that the degree of liposomes immune lysis depends on complement concentration in a wide range that can be used for definition of TCA level. Research of blood sera from patients has revealed correlation (r =0,793) between data received with the help of liposome immunolytic systems, and the data of nephelometric analysis with application of suspension sheep erythrocytes. Conclusion: The method allows to define total complement activity in blood serum in 15 minutes without separation of reaction components. This might be useful for measurement ТСА level at patients with various diseases and realization of scientific researches.

scholarly journals Study of hemolytic activity and subfactors of complement in smokers intoxication

2019 ◽  
Author(s):  
Rojan Ghanim Al Allaf
Keyword(s):  
Hemolytic Activity ◽  
Normal Values ◽  
Control Group ◽  
Healthy Individuals ◽  
Complement Components ◽  
Complement Activity ◽  
Normal People ◽  
Hemolytic Complement Activity ◽  
Heavy Smokers

Abstract Heavy Smokers appeared to be less resistant to infection, such as bacteria, viruses, and parasites. Many studies have examined the complement components concentrations than compared with normal people and ignored the functional sequencing of complement components, our study included the determination of complement activity by using Sheep red blood cells (SRBCs) as antigen and extracting the hemolytic activity (50%) of complement compounds, and because of difficulty of this method we using statistical analysis program (SPSS 23) and derived the inverse equation which gives the decomposition percentage (1-100%) of complement components by using five serum dilutions only. The total hemolytic complement activity (CH50) and its C3 and C4 fractions were determined in 30 heavy smokers. The results were compared with a control group that contained 30 persons matched in age and sex. Generally, both C3 and C4 concentrations were increased in smoker's individual in compared with the control group. However, when the independent t-test has applied the differences in the C3 and C4 levels in the control group (healthy individuals) and in the smoker group were found to be statistically insignificant but the inverse equation showed a 7% reduction in CH50 in smokers compared with the control group, where 18% reduction was observed. Our current study suggests that the complement components of the heavy smokers suffer from a significant dysfunction in the function, although the concentration of the basic components in the serum is parallel with normal values.

SOME FACTORS OF SIGNIFICANCE IN THE DOUBLE ANTIBODY IMMUNOASSAY OF INSULIN

1967 ◽  
Vol 54 (2) ◽  
pp. 347-361 ◽  
Author(s):  
K. Brunfeldt ◽  
K. R. Jorgensen
Keyword(s):  
Guinea Pig ◽  
Insulin Concentration ◽  
Precipitation Reaction ◽  
Antibody Technique ◽  
Complement Activity ◽  
Double Antibody ◽  
Insulin In Plasma ◽  
Antibody Method ◽  
Heparin Preparation

ABSTRACT Differences were demonstrated between 4 insulin antibody (guinea pig) precipitating antisera (rabbit), int. al. in their ability to cross react with human γ-globulin components. This cross reaction is presumably the reason why »negative« insulin values can be found at higher concentrations of certain precipitating antisera. Independently of the precipitating antiserum it was possible, in some cases, to demonstrate a difference between the insulin concentration in serum and in heparinized plasma. This difference could be eliminated by the addition of heparin, by long-lasting freezing, and by dilution of the serum. Hence, the higher values found in certain cases in serum, as compared with plasma are presumably due to the presence of complement. It is essential in the use of the double antibody technique for the determination of insulin in plasma and serum that a heparin preparation with anti-complement activity should be used and that the concentration range of the precipitating antiserum should be fixed. The double antibody method can be used with or without pre-precipitation if the above mentioned criteria are fulfilled. Pre-precipitation, however, is to be preferred, as the precipitation reaction here occurs under constant conditions.

Differences of Total Activity and Heat Tolerance of Complement in Plasma Between Black and Yellow Populations

2019 ◽  
Author(s):  
Abdelhakam G. Tamomh ◽  
Xiaojun Jin ◽  
Hui Liu
Keyword(s):  
Heat Tolerance ◽  
Total Activity ◽  
Ratio Method ◽  
Healthy Individuals ◽  
Blood Samples ◽  
Complement Activity ◽  
Temperature Calculation ◽  
Total Complement ◽  
Standard Reaction ◽  
Tolerance Distribution

Abstract Background The goal of this paper is to compare and evaluate the differences in total activity and Heat Tolerance of complement distribution Between Black and Yellow populations.Methods Blood samples of Black and Yellow healthy individuals were randomly collected, and plasma was obtained. The test plasma was diluted in a five series concentration, following with standard reaction and experimental reaction measurements. The basic principle of heat tolerance temperature calculation is to consider the ratio value (ratio method) in which heat tolerance of TCA calculated and plotted according to the area of trapezoidal and triangle as follows: C x°C=OD1-OD5+(OD2-OD5+OD3-OD5+OD4-OD5)*2. Where Cx°C is the total complement activity of the specimen measured in the two reaction conditions.Results The total activity of complement in Black individuals was statistically significantly lower than Yellow individuals (P< 0.05). The heat tolerance of total complement activity was higher among blacks than yellow individuals with no differences between the two groups (P> 0.05). The frequency values of total activity of complement distributed in the yellow individuals was statistically higher than Blacks (P<0.05) and The heat tolerance of total complement activity was statistically higher significant among Blacks than Yellow individuals (P<0.05).Conclusions There is a high differences in total complement activity and heat Tolerance distribution between Black and Yellow populations, may predict health status and innate immunity level differences between African race and Asian race.

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