scholarly journals Mumps Virus Nucleoprotein and Hemagglutinin-Specific Antibody Response Following a Third Dose of Measles Mumps Rubella Vaccine

2017 ◽  
Vol 4 (4) ◽  
Author(s):  
Donald R Latner ◽  
Amy Parker Fiebelkorn ◽  
Marcia McGrew ◽  
Nobia J Williams ◽  
Laura A Coleman ◽  
...  
Keyword(s):  
Control Measure ◽  
Neutralizing Antibody ◽  
Risk Groups ◽  
Mumps Virus ◽  
Enzyme Linked Immunosorbent Assay ◽  
Poor Correlation ◽  
Mmr Vaccine ◽  
Specific Antibody Response ◽  
Potential Benefits

Abstract Background Recent mumps outbreaks among 2-dose measles mumps rubella (MMR) vaccine recipients have raised questions regarding the potential benefits of a third dose of vaccine (MMR3). If MMR3 provides a sustained elevation in mumps antibody, it may be beneficial for certain at-risk groups or as an outbreak control measure. Methods Sera were collected immediately prior to MMR3 and at 1 month and 1 year post-MMR3 from 656 healthy adults aged 18–28 years in a nonoutbreak setting. Immunoglobulin G (IgG) was measured by enzyme-linked immunosorbent assay (ELISA) using whole mumps virus (commercial ELISA), hemagglutinin (HN; major neutralizing target), and nucleoprotein (NP; immunodominant) antigens. ELISA measurements were compared with in vitro plaque reduction neutralization (PRN) titers, and baseline antibody was compared with post-MMR3 levels. Results There were modest but statistically significant (P < .05) increases in mumps antibody at 1 month post-MMR3 by all 3 ELISA methods and by PRN titer. At 1 year post-MMR3, mumps antibody declined toward baseline but remained elevated (P < .05). The correlation between PRN titers and ELISA measurements was poor (r2 = .49), although sera with the highest amount of HN IgG also had the highest PRN titers. Conclusions Individuals with the lowest baseline PRN titers had the largest increase in frequency of samples that became positive for HN and NP by ELISA. A third dose of MMR may benefit certain individuals with a low level of mumps virus–neutralizing antibody, especially in the context of an outbreak or other high-risk setting. Additionally, poor correlation among serologic tests does not allow effective prediction of PRN titer by ELISA.

Autonomous Megakaryocyte Growth in Essential Thrombocythemia and Idiopathic Myelofibrosis Is Not Related to a c-mpl Mutation or to an Autocrine Stimulation by Mpl-L

Blood ◽  
1999 ◽  
Vol 93 (1) ◽  
pp. 125-139 ◽  
Author(s):  
Anne Laure Taksin ◽  
Jean-Pierre Le Couedic ◽  
Isabelle Dusanter-Fourt ◽  
Aline Massé ◽  
Stéphane Giraudier ◽  
...  
Keyword(s):  
Essential Thrombocythemia ◽  
Neutralizing Antibody ◽  
Inhibitory Effect ◽  
Enzyme Linked Immunosorbent Assay ◽  
Coding Region ◽  
Idiopathic Myelofibrosis ◽  
Spontaneous Growth ◽  
Cd34 Cells ◽  
Autocrine Stimulation

Abstract Essential thrombocythemia (ET) and idiopathic myelofibrosis (PMF) are two myeloproliferative diseases characterized by a marked megakaryocytic (MK) involvement. The pathogenesis of these two diseases is unknown. Recently it has been shown that overexpression of Mpl-ligand (Mpl-L) in mice induces thrombocytosis and myelofibrosis. In this study, we investigated whether Mpl-L was responsible for the pathogenesis of ET and PMF. Using in vitro cultures of blood or marrow CD34+ cells, we investigated whether MK growth was abnormal in these two diseases. Spontaneous MK growth involving only a fraction (20%) of the MK progenitors, as compared with growth in the presence of pegylated recombinant human megakaryocyte growth and development factor (PEG-rhuMGDF), was found in both diseases (21ET and 14PMF) using serum-free semisolid and liquid cultures, including cultures at one cell per well. We first searched for ac-mpl mutation/deletion by sequencing the entire coding region of the gene by polymerase chain reaction (PCR) in nine ET patients and five PMF patients, but no mutation was found. We subsequently investigated whether an autocrine stimulation by Mpl-L could explain the autonomous MK growth. Addition of different preparations of soluble Mpl receptor (sMpl) containing a Fc domain of IgG1 (sMpl-Fc) markedly inhibited MK spontaneous growth in both ET and PMF patients. This effect was specific for sMpl because a control soluble receptor (s4-1BB-Fc) had no inhibitory effect and an sMpl devoid of the Fc fragment had the same inhibitory efficacy as the sMpl-Fc. This inhibition was reversed by addition of PEG-rhuMGDF or a combination of cytokines. The sMpl-Fc markedly altered the entry into cell cycle of the CD34+ cells and increased the apoptosis that occurs in most patient CD34+ cells in the absence of exogenous cytokine, suggesting an autocrine stimulation. In contrast, a neutralizing antibody against Mpl-L did not alter the spontaneous MK growth, whereas it totally abolished the effects of 10 ng/mL PEG-rhuMGDF on patient or normal CD34+ cells. Mpl-L transcripts were detected at a very low level in the patient CD34+cells and MK and only when a highly sensitive fluorescent PCR technique was used. By quantitative reverse-transcription (RT)-PCR, the number of Mpl-L transcripts per actin transcripts was lower than detected in human Mpl-L–dependent cell lines, suggesting that this synthesis of Mpl-L was not biologically significant. In favor of this hypothesis, the Mpl-L protein was not detected in culture supernatants using either an enzyme-linked immunosorbent assay (ELISA) or a biological (Ba/F3huc-mpl) assay, except in one PMF patient. Investigation of Mpl-L signaling showed an absence of constitutive activation of STATs in spontaneously growing patient MKs. Addition of PEG-rhuMGDF to these MKs activated STATs 3 and 5. This result further suggests that spontaneous growth is neither related to a stimulation by Mpl-L nor to ac-mpl mutation. In conclusion, our results show that Mpl-L or Mpl are not directly implicated in the abnormal proliferation of MK cells from ET and PMF. The mechanisms by which the sMpl mediates a growth inhibition will require further experiments.

Glycated calmodulin from platelets as an index of glycemic control

1993 ◽  
Vol 39 (5) ◽  
pp. 815-819 ◽  
Author(s):  
A Muruganandam ◽  
G J Romsa ◽  
R J Thibert ◽  
R M Cheung ◽  
T F Draisey ◽  
...  
Keyword(s):  
Glycemic Control ◽  
Time Window ◽  
Human Subjects ◽  
Enzyme Linked Immunosorbent Assay ◽  
Poor Correlation ◽  
Type I ◽  
Clinical Role ◽  
Short Time Window ◽  
Short Time

Abstract In an effort to test whether a significant fraction of calmodulin would become glycated within the life span of the platelet (10-14 days), we monitored the kinetics of calmodulin glycation in vitro. Under the conditions we used, the fraction of glycated calmodulin reached a maximum (approximately 21%) within 10 days. We then extended the studies to human subjects. The intraplatelet concentrations of calmodulin and glycated calmodulin from age-matched type I diabetic subjects were monitored by a combination of m-aminophenylboronate affinity chromatography and enzyme-linked immunosorbent assay. The results indicate that the concentrations of total intraplatelet calmodulin (nonglycated plus glycated) were not dependent on the glycemic state of the subjects. Data from control and diabetic subjects showed a poor correlation between the concentrations of glycohemoglobin and of glycated calmodulin. However, a better correlation was obtained when glycated calmodulin concentrations were compared with those of serum fructosamine. The fraction of glycated calmodulin in the control population (7.71% +/- 0.75%) was significantly (P < 0.05) different from that of the diabetic population (21.6% +/- 1.26%). Given that the clinical role of the fructosamine assay remains controversial, estimation of glycated calmodulin in platelets might be useful as a short time-window index of glycemic control.

Autonomous Megakaryocyte Growth in Essential Thrombocythemia and Idiopathic Myelofibrosis Is Not Related to a c-mpl Mutation or to an Autocrine Stimulation by Mpl-L

Blood ◽  
1999 ◽  
Vol 93 (1) ◽  
pp. 125-139 ◽  
Author(s):  
Anne Laure Taksin ◽  
Jean-Pierre Le Couedic ◽  
Isabelle Dusanter-Fourt ◽  
Aline Massé ◽  
Stéphane Giraudier ◽  
...  
Keyword(s):  
Essential Thrombocythemia ◽  
Neutralizing Antibody ◽  
Inhibitory Effect ◽  
Enzyme Linked Immunosorbent Assay ◽  
Coding Region ◽  
Idiopathic Myelofibrosis ◽  
Spontaneous Growth ◽  
Cd34 Cells ◽  
Autocrine Stimulation

Essential thrombocythemia (ET) and idiopathic myelofibrosis (PMF) are two myeloproliferative diseases characterized by a marked megakaryocytic (MK) involvement. The pathogenesis of these two diseases is unknown. Recently it has been shown that overexpression of Mpl-ligand (Mpl-L) in mice induces thrombocytosis and myelofibrosis. In this study, we investigated whether Mpl-L was responsible for the pathogenesis of ET and PMF. Using in vitro cultures of blood or marrow CD34+ cells, we investigated whether MK growth was abnormal in these two diseases. Spontaneous MK growth involving only a fraction (20%) of the MK progenitors, as compared with growth in the presence of pegylated recombinant human megakaryocyte growth and development factor (PEG-rhuMGDF), was found in both diseases (21ET and 14PMF) using serum-free semisolid and liquid cultures, including cultures at one cell per well. We first searched for ac-mpl mutation/deletion by sequencing the entire coding region of the gene by polymerase chain reaction (PCR) in nine ET patients and five PMF patients, but no mutation was found. We subsequently investigated whether an autocrine stimulation by Mpl-L could explain the autonomous MK growth. Addition of different preparations of soluble Mpl receptor (sMpl) containing a Fc domain of IgG1 (sMpl-Fc) markedly inhibited MK spontaneous growth in both ET and PMF patients. This effect was specific for sMpl because a control soluble receptor (s4-1BB-Fc) had no inhibitory effect and an sMpl devoid of the Fc fragment had the same inhibitory efficacy as the sMpl-Fc. This inhibition was reversed by addition of PEG-rhuMGDF or a combination of cytokines. The sMpl-Fc markedly altered the entry into cell cycle of the CD34+ cells and increased the apoptosis that occurs in most patient CD34+ cells in the absence of exogenous cytokine, suggesting an autocrine stimulation. In contrast, a neutralizing antibody against Mpl-L did not alter the spontaneous MK growth, whereas it totally abolished the effects of 10 ng/mL PEG-rhuMGDF on patient or normal CD34+ cells. Mpl-L transcripts were detected at a very low level in the patient CD34+cells and MK and only when a highly sensitive fluorescent PCR technique was used. By quantitative reverse-transcription (RT)-PCR, the number of Mpl-L transcripts per actin transcripts was lower than detected in human Mpl-L–dependent cell lines, suggesting that this synthesis of Mpl-L was not biologically significant. In favor of this hypothesis, the Mpl-L protein was not detected in culture supernatants using either an enzyme-linked immunosorbent assay (ELISA) or a biological (Ba/F3huc-mpl) assay, except in one PMF patient. Investigation of Mpl-L signaling showed an absence of constitutive activation of STATs in spontaneously growing patient MKs. Addition of PEG-rhuMGDF to these MKs activated STATs 3 and 5. This result further suggests that spontaneous growth is neither related to a stimulation by Mpl-L nor to ac-mpl mutation. In conclusion, our results show that Mpl-L or Mpl are not directly implicated in the abnormal proliferation of MK cells from ET and PMF. The mechanisms by which the sMpl mediates a growth inhibition will require further experiments.

Detection of anti-HTLV-I Tax antibodies in HTLV-I enzyme-linked immunosorbent assay-negative individuals

Blood ◽  
1989 ◽  
Vol 74 (3) ◽  
pp. 1066-1072 ◽  
Author(s):  
GD Ehrlich ◽  
JB Glaser ◽  
MA Abbott ◽  
DJ Slamon ◽  
D Keith ◽  
...  
Keyword(s):  
High Risk ◽  
Immunosorbent Assay ◽  
Risk Groups ◽  
Enzyme Linked Immunosorbent Assay ◽  
Drug Abusers ◽  
Western Blots ◽  
True Incidence ◽  
Tax Protein ◽  
Radioimmunoprecipitation Assay

Abstract The HTLV-I tax gene protein (Tax) is not packaged within the mature viral particle from which the proteins for the commercially available enzyme-linked immunosorbent assay (ELISA) are derived. Screening of 162 individuals within a cohort of white intravenous (IV) drug abusers, previously identified as having an increased incidence of HTLV-I infection, demonstrated that seven of them had antibodies to the HTLV-I Tax protein but tested negative in HTLV-I ELISAs and Western blots prepared from purified virion proteins. Three out of 35 individuals in other behaviorally defined high-risk groups also displayed this limited pattern of reactivity to HTLV-I proteins. The presence of the anti-HTLV- I p40/Tax antibodies was determined by radioimmunoprecipitation assay (RIPA), which also revealed low levels of anti-env reactivity. The specificity of the anti-p40 reactivity was confirmed on specific Tax ELISAs and Western blots prepared from recombinantly produced Tax. In vitro gene amplification by the polymerase chain reaction (PCR) was used to establish the presence of sequences homologous to HTLV-I proviral DNA in four/four of these HTLV-I ELISA negative, Tax ELISA/Tax western blot/RIPA positive individuals. These data suggest that the true incidence of HTLV-I infection within high-risk cohorts is greater than previously reported.

scholarly journals Detection of anti-HTLV-I Tax antibodies in HTLV-I enzyme-linked immunosorbent assay-negative individuals

Blood ◽  
1989 ◽  
Vol 74 (3) ◽  
pp. 1066-1072
Author(s):  
GD Ehrlich ◽  
JB Glaser ◽  
MA Abbott ◽  
DJ Slamon ◽  
D Keith ◽  
...  
Keyword(s):  
High Risk ◽  
Immunosorbent Assay ◽  
Risk Groups ◽  
Enzyme Linked Immunosorbent Assay ◽  
Drug Abusers ◽  
Western Blots ◽  
True Incidence ◽  
Tax Protein ◽  
Radioimmunoprecipitation Assay

The HTLV-I tax gene protein (Tax) is not packaged within the mature viral particle from which the proteins for the commercially available enzyme-linked immunosorbent assay (ELISA) are derived. Screening of 162 individuals within a cohort of white intravenous (IV) drug abusers, previously identified as having an increased incidence of HTLV-I infection, demonstrated that seven of them had antibodies to the HTLV-I Tax protein but tested negative in HTLV-I ELISAs and Western blots prepared from purified virion proteins. Three out of 35 individuals in other behaviorally defined high-risk groups also displayed this limited pattern of reactivity to HTLV-I proteins. The presence of the anti-HTLV- I p40/Tax antibodies was determined by radioimmunoprecipitation assay (RIPA), which also revealed low levels of anti-env reactivity. The specificity of the anti-p40 reactivity was confirmed on specific Tax ELISAs and Western blots prepared from recombinantly produced Tax. In vitro gene amplification by the polymerase chain reaction (PCR) was used to establish the presence of sequences homologous to HTLV-I proviral DNA in four/four of these HTLV-I ELISA negative, Tax ELISA/Tax western blot/RIPA positive individuals. These data suggest that the true incidence of HTLV-I infection within high-risk cohorts is greater than previously reported.

scholarly journals Characterization of Alpha-ToxinhlaGene Variants, Alpha-Toxin Expression Levels, and Levels of Antibody to Alpha-Toxin in Hemodialysis and Postsurgical Patients with Staphylococcus aureus Bacteremia

2014 ◽  
Vol 53 (1) ◽  
pp. 227-236 ◽  
Author(s):  
Batu K. Sharma-Kuinkel ◽  
Yuling Wu ◽  
David E. Tabor ◽  
Hoyin Mok ◽  
Bret R. Sellman ◽  
...  
Keyword(s):  
Staphylococcus Aureus ◽  
Monoclonal Antibody ◽  
Clinical Outcome ◽  
Neutralizing Antibodies ◽  
Neutralizing Antibody ◽  
Enzyme Linked Immunosorbent Assay ◽  
Alpha Toxin ◽  
Content Type ◽  
Antibody Levels

Alpha-toxin is a majorStaphylococcus aureusvirulence factor. This study evaluated potential relationships betweenin vitroalpha-toxin expression ofS. aureusbloodstream isolates, anti-alpha-toxin antibody in serum of patients withS. aureusbacteremia (SAB), and clinical outcomes in 100 hemodialysis and 100 postsurgical SAB patients. Isolates underwentspatyping andhlasequencing. Serum anti-alpha-toxin IgG and neutralizing antibody levels were measured by using an enzyme-linked immunosorbent assay and a red blood cell (RBC)-based hemolysis neutralization assay. Neutralization of alpha-toxin by an anti-alpha-toxin monoclonal antibody (MAb MEDI4893) was tested in an RBC-based lysis assay. Most isolates encodedhla(197/200; 98.5%) and expressed alpha-toxin (173/200; 86.5%).In vitroalpha-toxin levels were inversely associated with survival (cure, 2.19 μg/ml, versus failure, 1.09 μg/ml;P< 0.01). Both neutralizing (hemodialysis, 1.26 IU/ml, versus postsurgical, 0.95;P< 0.05) and IgG (hemodialysis, 1.94 IU/ml, versus postsurgical, 1.27;P< 0.05) antibody levels were higher in the hemodialysis population. Antibody levels were also significantly higher in patients infected with alpha-toxin-expressingS. aureusisolates (P< 0.05). Levels of both neutralizing antibodies and IgG were similar among patients who were cured and those not cured (failures). Sequence analysis ofhlarevealed 12 distincthlagenotypes, and all genotypic variants were susceptible to a neutralizing monoclonal antibody in clinical development (MEDI4893). These data demonstrate that alpha-toxin is highly conserved in clinicalS. aureusisolates. Higherin vitroalpha-toxin levels were associated with a positive clinical outcome. Although patients infected with alpha-toxin-producingS. aureusexhibited higher anti-alpha-toxin antibody levels, these levels were not associated with a better clinical outcome in this study.

scholarly journals Sero-Prevalence of Rubella Virus among Pregnant Women in Kaduna State Nigeria 2015

2017 ◽  
Vol 9 (1) ◽  
Author(s):  
Aishatu Gubio ◽  
Steve Olonitola ◽  
Edward Jattau ◽  
Maryam Mukhtar
Keyword(s):  
Pregnant Women ◽  
Rubella Virus ◽  
Control Measure ◽  
First Trimester ◽  
Effective Vaccine ◽  
Enzyme Linked Immunosorbent Assay ◽  
Mmr Vaccine ◽  
Childbearing Age ◽  
Organ Systems ◽  
Positive Strand Rna

ObjectiveTo determine the IgM and IgG antibodies of rubella viruscirculating among pregnant women in Kaduna State Nigeria.IntroductionRubella virus causes -“German measles,” also known as “three-daymeasles.” This is usually a milder disease than red measles. Red/Hardmeasles or just measles is caused by Rubeola virus. The result of acuteinfection of the virus is a benign systematic rash which is significantlypathogenic to humans. This virus is a, positive-strand RNA virus thatreplicates in the cytoplasm of the infected cell.(Brooks et al., 2007).If placental infection of the virus spread during 8-10 weeks gestationit causes a chronic infection of the fetus leading to the developmentof congenital rubella syndrome (CRS) (Matthewset al., 2011) Theeffect of the infection of the several organ systems which include theeyes, ears, heart, brain, and endocrine system is known as congenitalrubella infection (CRI) (Chantleret, al.,2001)Rubella is endemic in Nigeria. Studies among women of childbearing age in Nigeria put seroprevalence at 66.6% in Imo, 77% inLagos and 93.5% in Oyo (8-10). Thus as part of the control measure,the availability of an effective vaccine to prevent Rubella infectionand therefore CRS, is necessary to evaluate the burden of disease ina country where MMR vaccine is not covered in the immunizationschedule or in vaccination strategyMethodsA cross-sectional study carried out on pregnant women attendingante-natal clinic from the three different senatorial district in Kadunastate. Blood samples were screened for rubella IgM & IgG antibodyusing commercially produced enzyme linked immunosorbent assay(ELISA), Questionnaires were administered to obtain demographicinformation and possible risk factors associated with rubella virus.Data was analzyed using Epi Info 6 Version 3.5.3.ResultsOf the 900 pregnant women screened, 572(63.3%) were positivefor rubella IgG. The prevalence of rubella IgG was highest among theage group 21-25 with 198(34.6%) and IgM was highest among theage group 21-25(51.3%). The IgG test results shows that 317 (66.0%)pregnant women tested positive for their first trimester, while the IgMpositive results shows 17(33.3%) for their first trimester. Although thesouthern senatorial district had the highest seroprevalence 14(35.9%)among the three centres, the differences were not statisticallysignificant (p>0.05). Only 3 people claimed to have been vaccinatedagainst rubella virus. Acquisition of primary education and being ahouse wife were insignificantly associated with raised titres. (p>0.05).ConclusionsThe serological evidence of rubella virus found in pregnant womenamong age group & their first trimester in this study is an indicationthat rubella is prevalent in Nigeria. It is however still necessary toimmunize seronegative women against rubella before they getpregnant.

scholarly journals Decreased humoral immunity to mumps in young adults immunized with MMR vaccine in childhood

2019 ◽  
Vol 116 (38) ◽  
pp. 19071-19076 ◽  
Author(s):  
Mohammed Ata Ur Rasheed ◽  
Carole J. Hickman ◽  
Marcia McGrew ◽  
Sun Bae Sowers ◽  
Sara Mercader ◽  
...  
Keyword(s):  
Young Adults ◽  
Humoral Immunity ◽  
Close Contact ◽  
Geometric Mean ◽  
Neutralizing Antibody ◽  
The United States ◽  
Enzyme Linked Immunosorbent Assay ◽  
Wild Type ◽  
Contributing Factors ◽  
Mmr Vaccine

In the past decade, multiple mumps outbreaks have occurred in the United States, primarily in close-contact, high-density settings such as colleges, with a high attack rate among young adults, many of whom had the recommended 2 doses of mumps-measles-rubella (MMR) vaccine. Waning humoral immunity and the circulation of divergent wild-type mumps strains have been proposed as contributing factors to mumps resurgence. Blood samples from 71 healthy 18- to 23-year-old college students living in a non-outbreak area were assayed for antibodies and memory B cells (MBCs) to mumps, measles, and rubella. Seroprevalence rates of mumps, measles, and rubella determined by IgG enzyme-linked immunosorbent assay (ELISA) were 93, 93, and 100%, respectively. The index standard ratio indicated that the concentration of IgG was significantly lower for mumps than rubella. High IgG avidity to mumps Enders strain was detected in sera of 59/71 participants who had sufficient IgG levels. The frequency of circulating mumps-specific MBCs was 5 to 10 times lower than measles and rubella, and 10% of the participants had no detectable MBCs to mumps. Geometric mean neutralizing antibody titers (GMTs) by plaque reduction neutralization to the predominant circulating wild-type mumps strain (genotype G) were 6-fold lower than the GMTs against the Jeryl Lynn vaccine strain (genotype A). The majority of the participants (80%) received their second MMR vaccine ≥10 years prior to study participation. Additional efforts are needed to fully characterize B and T cell immune responses to mumps vaccine and to develop strategies to improve the quality and durability of vaccine-induced immunity.

scholarly journals Estimates of Mumps Seroprevalence May Be Influenced by Antibody Specificity and Serologic Method

2013 ◽  
Vol 21 (3) ◽  
pp. 286-297 ◽  
Author(s):  
Donald R. Latner ◽  
Marcia McGrew ◽  
Nobia J. Williams ◽  
Sun B. Sowers ◽  
William J. Bellini ◽  
...  
Keyword(s):  
Neutralizing Antibodies ◽  
Neutralizing Antibody ◽  
Virus Antigen ◽  
Mumps Virus ◽  
Poor Correlation ◽  
Serologic Method ◽  
Specific Igg ◽  
Low Levels ◽  
The Individual ◽  
Antibody Levels

ABSTRACTNeutralizing antibodies are assumed to be essential for protection against mumps virus infection, but their measurement is labor- and time-intensive. For this reason, enzyme-linked immunosorbent assays (ELISAs) are typically used to measure mumps-specific IgG levels. However, since there is poor correlation between mumps neutralization titers and ELISAs that measure the presence of mumps-specific IgG levels, ELISAs that better correlate with neutralization are needed. To address this issue, we measured mumps antibody levels by plaque reduction neutralization, by a commercial ELISA (whole-virus antigen), and by ELISAs specific for the mumps nucleoprotein and hemagglutinin. The results indicate that differences in the antibody response to the individual mumps proteins could partially explain the lack of correlation among various serologic tests. Furthermore, the data indicate that some seropositive individuals have low levels of neutralizing antibody. If neutralizing antibody is important for protection, this suggests that previous estimates of immunity based on whole-virus ELISAs may be overstated.

An Enzyme Immunoassay (ELISA) for the Quantitation of Human Factor VII

1986 ◽  
Vol 56 (03) ◽  
pp. 250-255 ◽  
Author(s):  
C Boyer ◽  
M Wolf ◽  
C Rothschild ◽  
M Migaud ◽  
J Amiral ◽  
...  
Keyword(s):  
Human Factor ◽  
Solid Phase ◽  
Factor Vii ◽  
Enzyme Linked Immunosorbent Assay ◽  
Poor Correlation ◽  
Vein Thrombosis ◽  
Coagulant Activity ◽  
Significant Difference ◽  
Large Populations ◽  
Deep Vein

SummaryA new solid phase enzyme-linked immunosorbent assay (ELISA) was developed for the quantitation of human Factor VII antigen (F VII Ag), using a monospecific rabbit anti-F VII antiserum. Anti-F VII F(ab′)2 fragments were adsorbed to polystyrene plates. The binding of serial dilutions of control or test plasma, containing F VII, was detected by incubation with peroxidase-labeled anti- FV II IgG followed by the addition of hydrogen peroxyde and O-phenylenediamine. This ELISA is specific, sensitive (detection limit: 0.05%) and accurate (coefficient of variation: 1.5-4% for within- and 1.6-9% for between-assays). F VII coagulant activity (F VII C) and F VII Ag were determined in large populations of controls and patients. In normal plasma (n = 38), F VII Ag ranged from 83 to 117% and the correlation coefficient between F VII Ag and F VII C was 0.94. In patients with severe (F VII C inf. 1%) congenital F VII deficiency (n = 5), F VII Ag was undetectable in two cases (inf. 0.05%) and markedly reduced (0.35 to 5.6%) in the three other cases. In patients with liver cirrhosis (n = 15), F VII Ag ranged from 21 to 59% and was in good correlation with F VII C (r = 0.84). In dicoumarol treated patients (n = 15), the levels of F VII Ag ranged from 51% to 79% and a poor correlation (r = 0.52) with F VIIC was observed. In “compensated” DIC (n = 5), levels of F VII Ag varied from 60 to 186%, with significantly higher F VII C levels (from 143 to 189%). In contrast, in “decompensated” DIC (n = 7), low F VII Ag and F VII C levels were observed (from 7 to 27%). In patients with deep-vein thrombosis (n = 25), high levels of F VII Ag (from 102 to 136%) and F VII C (from 110 to 150%) were demonstrated. In surgical patients, no significant difference was observed before and one day after intervention.

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