scholarly journals An Immunoblot Assay for Detection of Immunoglobulin M Antibody to Human Herpesvirus 6

2000 ◽  
Vol 7 (5) ◽  
pp. 823-827 ◽  
Author(s):  
Steve LaCroix ◽  
John A. Stewart ◽  
Margaret E. Thouless ◽  
Jodi B. Black
Keyword(s):  
Human Serum ◽  
Acute Phase ◽  
Human Herpesvirus ◽  
Immunoglobulin M ◽  
Human Herpesvirus 6 ◽  
Serum Samples ◽  
Immunoblot Assay ◽  
Human Serum Samples ◽  
Convalescent Phase ◽  
Phase Sample

ABSTRACT We identified the human herpesvirus 6 (HHV-6)-dominant immunoglobulin M (IgM)-reactive virion protein as being the same 101-kDa protein (101K) previously identified as the major IgG immunoreactive protein and a specific serologic marker of HHV-6 infection. An immunoblot assay (IB) to detect HHV-6-specific IgM antibodies against the 101K protein in human serum samples was developed. The assay was validated by using acute- and convalescent-phase serum collected from children under 2 years of age in which we previously detected IgG seroconversion to the HHV-6 101K protein. Of 32 serum pairs which previously demonstrated IgG seroconversion to the 101K protein, 29 had IgM reactivity to the same protein in the acute-phase sample and the remaining 3 had reactivity in the convalescent-phase sample. We also detected HHV-6 IgM activity in sera collected from individuals ≥4 years of age who were also IgM seropositive to measles or rubella. Results of cross-adsorption studies using measles virus-, rubella virus-, and HHV-6-infected cells as the adsorbing antigen indicated no cross-reactivity between measles or rubella IgM and HHV-6 IgM in human serum samples. The IgM IB detected HHV-6-specific IgM antibody to the 101K protein in 78% (63 of 81) of tested acute-phase serum collected from young children with an undifferentiated rash illness by using a single serum dilution.

scholarly journals Serum Antibody Responses in Children with Rotavirus Diarrhea Can Serve as Proxy for Protection

2005 ◽  
Vol 12 (2) ◽  
pp. 273-279 ◽  
Author(s):  
J. Xu ◽  
P. Dennehy ◽  
H. Keyserling ◽  
L. E. Westerman ◽  
Y. Wang ◽  
...  
Keyword(s):  
Acute Phase ◽  
Severe Disease ◽  
Serum Antibody ◽  
Immunoglobulin M ◽  
Antibody Responses ◽  
Serum Samples ◽  
Convalescent Phase ◽  
Conversion Rates ◽  
Reliable Marker ◽  
Rotavirus Diarrhea

ABSTRACT We examined sera from 42 patients 1 to 30 months of age for rotavirus immunoglobulin M (IgM), IgA, IgG, and IgG subclasses and sought to determine if serum antibody could serve as a reliable marker for prediction of disease severity. Infants in the first few months of life usually had high maternal IgG titers and, when they were infected with rotavirus, had low IgM titers or no IgM in acute-phase sera and poor seroconversions 3 weeks later, suggesting that maternal antibodies had inhibited viral replication and antibody responses. All patients ≥6 months of age had IgM in acute-phase sera, indicating that IgM is a good marker for acute rotavirus infection. IgG was the best overall predictor of an infection, as the convalescent-phase sera of 81% of the patients had a fourfold rise in the IgG titer. IgA titers in convalescent-phase sera and conversion rates were higher among patients ≥12 months of age than among children younger than 12 months. IgG1 was the predominant subclass detected in the acute-phase sera of some children and in all 28 convalescent-phase serum samples examined. Patients with preexisting acute-phase IgG titers of ≥100 or ≥200 had diarrhea that was less severe or of a shorter duration. These results indicate that serum IgG is the most reliable marker for seroconversion and is a consistent proxy for protection against severe disease.

scholarly journals Crimean-Congo Hemorrhagic Fever in Iraq During 2010

2012 ◽  
Vol 36 (0E) ◽  
pp. 99-103
Author(s):  
Emad S. Abul-Eis ,
Keyword(s):  
Human Serum ◽  
Hemorrhagic Fever ◽  
Zoonotic Disease ◽  
Immunoglobulin M ◽  
Serum Samples ◽  
Crimean Congo Hemorrhagic Fever ◽  
Human Serum Samples ◽  
Specific Immunoglobulin ◽  
Cchf Virus

Crimean-Congo hemorrhagic fever (CCHF) is a viral zoonotic disease with a highmortality rate in humans. CCHF is caused by genus Nairovirus, in family of Bunyaviridae,and is transmitted to humans through the bite of ticks Hyalomma spp or contact with blood ortissues of CCHF patients or infected livestock.The total numbers of positive patients to CCHF virus was 11 out of 44 suspected sampleswere examined from eight provinces during the period from January to December 2010 . Theway of transmission is due to contact with blood and tissues of infected animals, and onepatient slaughtered sheep in his house. ELISA was used to detect Crimean-Congohemorrhagic fever (CCHF) virus-specific immunoglobulin M (IgM) in human serum samples.

scholarly journals Analysis of Eight Commercial Enzyme Immunoassay Tests for Detection of Antibodies to Mycoplasma pneumoniae in Human Serum

2004 ◽  
Vol 11 (5) ◽  
pp. 862-867 ◽  
Author(s):  
Deborah F. Talkington ◽  
Susan Shott ◽  
Michael T. Fallon ◽  
Stephanie B. Schwartz ◽  
W. Lanier Thacker
Keyword(s):  
Mycoplasma Pneumoniae ◽  
Acute Phase ◽  
The United States ◽  
Immunoglobulin M ◽  
Atypical Pneumonia ◽  
Serum Samples ◽  
Convalescent Phase ◽  
Single Use ◽  
Positive Results ◽  
Plate Type

ABSTRACT Mycoplasma pneumoniae is an important etiologic agent of primary atypical pneumonia in children and adults. The diagnosis of M. pneumoniae infection is commonly confirmed through serologic testing. In this study, we used paired sera from 51 patients (all with confirmed M. pneumoniae infection and positive complement fixation [CF] titers) to compare the results of eight enzyme immunoassays (EIAs) available commercially in the United States. We compared two single-use EIAs and six plate-type EIAs. Results from acute-phase sera ranged from only 7 (14%) positive by ImmunoWELL (GenBio) immunoglobulin M (IgM) EIA to 23 (45%) positive by Zeus IgG EIA. When both the acute-phase and convalescent-phase serum samples were examined, positive results ranged from 20 (39%) by the ImmunoWELL (GenBio) IgM assay to 45 (88%) positive by the Remel IgG-IgM EIA. In this study, the single-use EIAs by Remel and Meridian were more reliable than were the plate-type EIAs. Among the plate-type EIAs, the Zeus and DiaSorin assays (which detect antibodies to protein antigens) were more sensitive than the ImmunoWELL assay (which detects antibodies to glycolipid antigens). In general, IgG EIAs on convalescent-phase sera were more concordant with one another than were IgM EIAs with one another. Scatter plot analysis of convalescent-phase sera showed that, as the CF titer dropped, the IgM assays identified fewer positive convalescent-phase sera. In contrast, the IgG assays provided fairly consistent positive results for convalescent-phase sera with CF titers of 64 and above. Results of individual tests and overall limitations of serodiagnostics for M. pneumoniae infections are discussed.

scholarly journals Diagnosis of Mycoplasma pneumoniaePneumonia in Children

1998 ◽  
Vol 36 (11) ◽  
pp. 3155-3159 ◽  
Author(s):  
Matti E. Waris ◽  
Pia Toikka ◽  
Taina Saarinen ◽  
Simo Nikkari ◽  
Olli Meurman ◽  
...  
Keyword(s):  
Acute Phase ◽  
Southern Hybridization ◽  
Solid Phase ◽  
Laboratory Diagnosis ◽  
Immunoglobulin M ◽  
Serum Samples ◽  
Convalescent Phase ◽  
Pcr Products ◽  
Serology Test ◽  
Time Of Admission

We evaluated a commercial immunoglobulin M (IgM)-capture immunoassay for the detection of Mycoplasma pneumoniaeinfections in 278 pediatric patients with community-acquired, radiographically defined pneumonia. Acute- and convalescent-phase serum samples were collected from all patients and were tested for M. pneumoniae-specific IgM and IgG antibodies by Platelia enzyme immunoassays (Sanofi Diagnostica Pasteur, Marnes la Coquette, France). Nasopharyngeal aspirates (NPAs) were collected at the time of admission to the hospital. A total of 227 NPAs were subjected to the detection ofM. pneumoniae DNA by PCR, and 191 NPAs were cultured by using the Pneumofast kit (International Mycoplasma, Signeswere, France). Southern hybridization of PCR products and the IgM test with solid-phase antigen (Serion Immunodiagnostica, Würzburg, Germany) were used for additional confirmation of a positive result, which required agreement of at least two different methods. A total of 24 (9%) confirmed diagnoses of mycoplasma infection were made, 5 (21%) of which were in children <5 years of age. Of the positive children, 24 of 24 (sensitivity, 100%) were positive by the IgM-capture test with convalescent-phase serum, 19 of 24 (79%) were positive by the IgM-capture test with acute-phase serum, 19 of 24 (79%) were positive by IgG serology, 10 of 20 (50%) were positive by PCR, and 8 of 17 (47%) were positive by culture. An additional 5 (of 254) children were positive by the Platelia IgM test alone (specificity, 98%). When the PCR with Southern hybridization result was combined with the IgM-capture test result with the acute-phase sera, the sensitivity of rapid laboratory diagnosis increased to 95%. In conclusion, the IgM serology test was the single most valuable tool for the diagnosis ofM. pneumoniae pneumonia in children of any age.

Simultaneous Monitoring of Febuxostat and Uric Acid in Human Serum Samples Using the Direct Square-Wave Voltammetric Method

2019 ◽  
Vol 15 (6) ◽  
pp. 678-684
Author(s):  
Biljana Nigović ◽  
Jakov Vlak
Keyword(s):  
Uric Acid ◽  
Human Serum ◽  
Oxidase Inhibitor ◽  
Serum Samples ◽  
Boron Doped Diamond ◽  
Fast Analysis ◽  
Xanthine Oxidase Inhibitor ◽  
Voltammetric Method ◽  
Square Wave ◽  
Human Serum Samples

Background: High uric acid serum level, hyperuricemia, is now associated with many diseases such as gout, chronic kidney disease, hypertension, coronary artery disease and diabetes. Febuxostat is a novel selective xanthine oxidase inhibitor approved for the treatment of hyperuricemia. Objective: The aim of this study was to develop a first analytical method for the simultaneous determination of febuxostat and uric acid. Methods: An unmodified boron-doped diamond electrode provided concurrent quantitation of drug at low levels and uric acid, which has clinical significance in the diagnosis and therapy of hyperuricemia, at relatively high concentrations. The direct square-wave voltammetric method was applied to the analysis of both analytes in human serum samples. Results: Under the optimized conditions, the linear response of peak current on febuxostat concentration was achieved in the range from 7.5 × 10-7 to 3 × 10-5 M, while uric acid showed two linear ranges of 5 × 10-6 - 5 × 10-5 M and 5 × 10-5 - 2 × 10-4 M. The method was successfully utilised for quantification of both analytes in human serum samples. Good recoveries were obtained without interference from common inorganic cations and anions as well as glucose, dopamine, ascorbic and folic acids at concentrations expected in physiological conditions. Conclusion: The great benefits of developed method are fast analysis (only 7.5 s for run), low cost and simplicity of performance.

A rat granulosa cell plasminogen activator bioassay for FSH in human serum

1990 ◽  
Vol 126 (1) ◽  
pp. 159-168 ◽  
Author(s):  
A. N. Thakur ◽  
R. Coles ◽  
A. Sesay ◽  
B. Earley ◽  
H. S. Jacobs ◽  
...  
Keyword(s):  
Human Serum ◽  
Plasminogen Activator ◽  
Granulosa Cell ◽  
Serum Samples ◽  
Glycoprotein Hormone ◽  
Assay Sensitivity ◽  
Serum Factors ◽  
Human Serum Samples ◽  
Heat Treated ◽  
Fsh Bioactivity

ABSTRACT A previously described in-vitro rat granulosa cell plasminogen activator bioassay for FSH has been modified and applied in the assay of human serum. This modified method consists of exposing the diethylstilboestrol-stimulated granulosa cells from 25- to 26-day-old rats to FSH or test substance for 3·5 h in wells coated with 125I-labelled fibrinogen and treated with thrombin. Following stimulation with FSH, the dose-related production of plasminogen activator was measured as the degree of 125I-labelled fibrinolysis in the presence of added plasminogen. Using the urinary FSH/LH bioassay reference preparation as the assay standard, the useful range of the assay was 0·3–15IU/l, with an assay sensitivity of 0·3 IU/l. As determined using purified glycoprotein hormone preparations, the assay was highly specific for FSH. The minor degree of FSH bioactivity measured in some of the hormone preparations was accounted for by the amount of FSH contamination in these preparations. To abolish interference caused by unknown serum factors, we heat-treated the serum samples for 15 min at 56 °C before the assay. The results indicated that neither immunoreactivity nor bioactivity was affected by this treatment. Furthermore, heat-treated human sera gave responses parallel to the standard curve at the three dose levels (2, 4 and 8 μl) studied. We used this bioassay to estimate the FSH-like bioactivity in 15 human serum samples. The estimates of immunoreactive FSH in these samples correlated well with the corresponding FSH bioactivity (r = 0·745, n = 15 and P < 0·05). The results indicate that with this sensitive and rapid (completed within 24 h) bioassay, it should be possible to measure FSH bioactivity in heat-treated human serum samples. Journal of Endocrinology (1990) 126, 159–168

scholarly journals Evaluation of PCR-based assay in human serum samples for diagnosis of fatal cases of spotted fever group rickettsiosis

2009 ◽  
Vol 15 ◽  
pp. 232-234 ◽  
Author(s):  
E.M Mendes do Nascimento ◽  
S. Colombo ◽  
T.K. Nagasse-Sugahara ◽  
R.N. Angerami ◽  
M.R. Resende ◽  
...  
Keyword(s):  
Human Serum ◽  
Spotted Fever ◽  
Spotted Fever Group ◽  
Serum Samples ◽  
Human Serum Samples

Simultaneous determination of ascorbic and uric acids and dopamine in human serum samples using three-way calibration with data from square wave voltammetry

2016 ◽  
Vol 129 ◽  
pp. 205-212 ◽  
Author(s):  
Adrian Marcelo Granero ◽  
Gastón Darío Pierini ◽  
Sebastián Noel Robledo ◽  
María Susana Di Nezio ◽  
Héctor Fernández ◽  
...  
Keyword(s):  
Human Serum ◽  
Simultaneous Determination ◽  
Square Wave Voltammetry ◽  
Serum Samples ◽  
Square Wave ◽  
Human Serum Samples ◽  
Wave Voltammetry

scholarly journals A Novel, Rapid, and Low-Volume Assay for Therapeutic Drug Monitoring of Posaconazole and Other Long-Chain Azole-Class Antifungal Drugs

mSphere ◽  
2018 ◽  
Vol 3 (6) ◽  
Author(s):  
Gregory R. Wiedman ◽  
Yanan Zhao ◽  
David S. Perlin
Keyword(s):  
Liquid Chromatography ◽  
Therapeutic Drug Monitoring ◽  
Human Serum ◽  
Drug Monitoring ◽  
Fungal Infections ◽  
Antifungal Drugs ◽  
Therapeutic Drug ◽  
Serum Samples ◽  
Human Serum Samples ◽  
Long Chain

ABSTRACT Clinicians need a better way to accurately monitor the concentration of antimicrobials in patient samples. In this report, we describe a novel, low-sample-volume method to monitor the azole-class antifungal drug posaconazole, as well as certain other long-chain azole-class antifungal drugs in human serum samples. Posaconazole represents an important target for therapeutic drug monitoring (TDM) due to its widespread use in treating invasive fungal infections and well-recognized variability of pharmacokinetics. The current “gold standard” requires trough and peak monitoring through high-pressure liquid chromatography (HPLC) or liquid chromatography-tandem mass spectroscopy (LC-MS/MS). Other methods include bioassays that use highly susceptible strains of fungi in culture plates or 96-well formats to monitor concentrations. Currently, no method exists that is both highly accurate in detecting free drug concentrations and is also rapid. Herein, we describe a new method using reduced graphene oxide (rGO) and a fluorescently labeled aptamer, which can accurately assess clinically relevant concentrations of posaconazole and other long-chain azole-class drugs in little more than 1 h in a total volume of 100 µl. IMPORTANCE This work describes an effective assay for TDM of long-chain azole-class antifungal drugs that can be used in diluted human serum samples. This assay will provide a quick, cost-effective method for monitoring concentrations of drugs such as posaconazole that exhibit well-documented pharmacokinetic variability. Our rGO-aptamer assay has the potential to improve health care for those struggling to treat fungal infections in rural or resource-limited setting.

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