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.

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 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 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 Detection of Bovine Herpesvirus-1-Specific IgM Using a Capture Enzyme Immune Assay with Isotype-Specific Monoclonal Antibodies

1989 ◽  
Vol 1 (2) ◽  
pp. 139-145 ◽  
Author(s):  
Fernando Osorio ◽  
Subramaniam Srikumaran ◽  
Marvin Rhodes ◽  
David Christensen ◽  
Pushpa Srikumaran
Keyword(s):  
Monoclonal Antibodies ◽  
Solid Phase ◽  
Bovine Herpesvirus ◽  
Immunoglobulin M ◽  
Viral Reactivation ◽  
Clinical Samples ◽  
Serum Samples ◽  
Bovine Herpesvirus 1 ◽  
Igm Antibody ◽  
Herpesvirus 1

The detection of virus-specific immunoglobulin M (IgM) antibodies in acute-phase serum samples offers the possibility of making an accurate and rapid serologic diagnosis. We have developed a solid-phase capture assay that uses murine monoclonal antibodies specific for bovine IgM to separate the whole IgM fraction of a bovine serum sample. The IgM specific for bovine herpesvirus-1 (BHV-1) is then detected by the addition of viral antigen, which in turn is detected by BHV-1-specific monoclonal antibodies conjugated to horseradish peroxidase. A BHV-1 IgM antibody response was detected during the early postinfection period (7–40 days PI). Bovine herpesvirus-1 IgM antibody was not detected in sera taken from 3 animals following dexamethasone-induced viral reactivation. This method compares favorably with viral isolation, antigen detection in the clinical samples, and paired serology in the diagnosis of BHV-1 infection at a herd level.

scholarly journals Laboratory Diagnosis of Toscana Virus Infection by Enzyme Immunoassay with Recombinant Viral Nucleoprotein

1999 ◽  
Vol 37 (3) ◽  
pp. 649-652 ◽  
Author(s):  
Dario Soldateschi ◽  
Grazia Maria Dal Maso ◽  
Marcello Valassina ◽  
Laura Santini ◽  
Silvia Bianchi ◽  
...  
Keyword(s):  
Enzyme Immunoassay ◽  
Nested Pcr ◽  
Immune Status ◽  
Laboratory Diagnosis ◽  
Summer Season ◽  
Immunoglobulin M ◽  
Toscana Virus ◽  
Serum Samples ◽  
Rna Sequences ◽  
Specific Serum

A recombinant enzyme immunoassay (rEIA) to detect serum immunoglobulin M (IgM) and IgG to Toscana virus (TOSV) was developed with the aim of establishing a simple and easily available assay for diagnosing acute and/or previous infections. The rEIA, based on the recombinant nucleoprotein of TOSV expressed in Escherichia coli, was evaluated with 97 serum samples collected in an area where TOSV is endemic and compared to an analogous assay based on cell-derived TOSV. Discordant results were resolved by immunoblotting (IB). Twenty-two of these samples, obtained from subjects hospitalized during the summer season with meningitis of suspected TOSV etiology, were further characterized by indirect immunofluorescence and IB, and detection of specific TOSV RNA sequences in the cerebrospinal fluid of these patients was attempted by nested PCR. The results indicated that rEIA was able to diagnose acute TOSV infection by detection of specific serum IgM in all of the subjects with TOSV meningitis confirmed by nested PCR or serology. The overall sensitivity and specificity of rEIA were both 100% for IgM detection and 100 and 96.6%, respectively, for IgG detection. Thus, rEIA appears to be a simple and reliable laboratory test for the diagnosis of acute TOSV infection and for the assessment of immune status.

Evaluation of Visceral Larva Migrans by Radioimmunoassay Systems

PEDIATRICS ◽  
1975 ◽  
Vol 56 (3) ◽  
pp. 417-420
Author(s):  
Roy Patterson ◽  
Mary Roberts ◽  
Robert J. Hart ◽  
Carolyn C. Huntley
Keyword(s):  
Acute Phase ◽  
Solid Phase ◽  
Larva Migrans ◽  
Parasitic Diseases ◽  
Igg Antibodies ◽  
Visceral Larva Migrans ◽  
Serum Samples ◽  
Solid Phase Radioimmunoassay ◽  
Antibody Levels ◽  
Hyperimmunoglobulinemia E

A 9-year-old child with miliary pulmonary infiltrates, eosinophilia, and hyperimmunoglobulinemia E recovered rapidly over a four-week period. Subsequent analysis of serum samples by a solid phase radioimmunoassay technique demonstrated IgM, IgE, and IgG antibodies to Ascaris suum antigen which declined following the acute phase of the illness in parallel with a decline in serum IgM, IgE, and IgG concentrations. Precipitating antibodies in serum against Ascaris antigen were demonstrated. The diagnosis is considered to be toxocariasis or ascariasis. The application of sensitive radioimmunoassay techniques of this type should provide a method of earlier diagnosis and the demonstration of rapidly changing antibody levels a method of confirming the diagnosis in parasitic diseases.

scholarly journals Quantitative PCR-Enhanced Immunoassay for Measurement of Enteroviral Immunoglobulin M Antibody and Diagnosis of Aseptic Meningitis

2005 ◽  
Vol 12 (2) ◽  
pp. 235-241 ◽  
Author(s):  
Amal Elfaitouri ◽  
Nahla Mohamed ◽  
Jan Fohlman ◽  
Robert Aspholm ◽  
Gun Frisk ◽  
...  
Keyword(s):  
Aseptic Meningitis ◽  
Solid Phase ◽  
Immunoglobulin M ◽  
Serum Samples ◽  
Serum Dilution ◽  
Group 4 ◽  
Control Serum ◽  
Group 3 ◽  
Group 2 ◽  
Capture Technique

ABSTRACT A PCR-enhanced immunoassay (PIA) to detect enterovirus (EV) immunoglobulin M (IgM) for diagnosis of recent EV infection was recently developed. This test was compared with another EV IgM capture technique, the solid-phase reverse immunosorbent test (SPRIST). Fourteen of 43 serum samples from aseptic meningitis patients were positive by PIA, whereas 10 were positive by SPRIST. One of 39 control serum samples was weakly positive by PIA. A single-serum-dilution real-time PCR-based PIA for EV IgM (quantitative PIA [QPIA]) was also developed and evaluated against PIA, SPRIST, an EV IgM radioimmunoassay (RIA), and clinical data. A mixture of 12 EVs was used as the antigen. Results from investigating four groups of serum samples were as follows. (i) The nine PIA-positive serum samples in group 1 were all positive by QPIA. (ii) Group 2 consisted of 59 serum samples from aseptic meningitis patients. Nineteen of 30 serum samples (63%) taken at hospital admission were positive by QPIA. Of these, 17 were positive in EV PCR. (iii) None of the 30 control serum samples in group 3 were positive by QPIA. (iv) For the 24 serum samples in group 4, of which 11 were positive and 13 were negative by RIA, the QPIA results were completely concordant. The sensitivity and specificity of QPIA for diagnosis of EV infection were 70 and 80%, respectively. QPIA provides a rational strategy for the detection of EV IgM, allows the use of viral antigens with minimal purification, and needs no virus-specific reagents apart from those in the PCR. QPIA is a generally applicable method for the detection of viral IgM in IgM capture assays.

scholarly journals Performance of Indirect Immunoglobulin M (IgM) Serology Tests and IgM Capture Assays for Laboratory Diagnosis of Measles

2000 ◽  
Vol 38 (1) ◽  
pp. 99-104
Author(s):  
Samuel Ratnam ◽  
Graham Tipples ◽  
Carol Head ◽  
Micheline Fauvel ◽  
Margaret Fearon ◽  
...  
Keyword(s):  
Measles Virus ◽  
False Negative ◽  
Laboratory Diagnosis ◽  
Immunoglobulin M ◽  
Confirmatory Test ◽  
Serum Samples ◽  
Negative Results ◽  
Capture Assay ◽  
Specimen Collection ◽  
Control And Prevention

ABSTRACT As progress is made toward elimination of measles, the laboratory confirmation of measles becomes increasingly important. However, both false-positive and false-negative results can occur with the routinely used indirect measles immunoglobulin M (IgM) serology tests. The measles IgM capture assay is considered to be more specific, and therefore, its use is indicated for confirmatory testing, but its relative performance has not been fully assessed. Four commercial indirect measles IgM serology test kits (the Behring, Clark, Gull, and PanBio assays) and a commercial IgM capture assay (the Light Diagnostics assay) were evaluated for their abilities to detect measles virus-specific IgM antibody with a total of 308 serum samples from patients involved in a measles outbreak and with confirmed cases of measles and 454 samples from subjects without measles. The Centers for Disease Control and Prevention (CDC) IgM capture assay was also used in a part of the evaluation. Among the indirect assays, the overall sensitivities ranged from 82.8% (Clark assay) to 88.6% (Behring assay) and specificity ranged from 86.6% (PanBio assay) to 99.6% (Gull assay). These rates were 92.2 and 86.6%, respectively, for the Light Diagnostics capture assay and 87.0 and 94.8%, respectively, for the CDC capture assay. While the Light Diagnostics capture assay had the best detection rate (80%) with the acute-phase samples compared with those for the rest of the tests (CDC capture assay, 77%; Behring assay, 70%; Gull assay, 69%; PanBio assay, 58%; and Clark assay, 57%), all tests showed a significantly improved sensitivity in the range of 92% (Clark and PanBio assays) to 97% (Light Diagnostics and CDC capture assays) with the convalescent-phase samples, as expected. The best seropositivity rates (in the range of 92 to 100%) were observed with samples collected 6 to 14 days after the onset of symptoms. The Gull assay showed the highest positive predictive value (99.6%), followed by the Behring assay (97.8%) and the CDC capture assay (96.1%). Overall, the Gull and Behring assays were found to be as good as or better than the capture assays. In conclusion, laboratory diagnosis of measles based on IgM serology varies depending on the timing of specimen collection and the test used, and the case for the use of the IgM capture assay as the confirmatory test appears to be uncertain.

scholarly journals Real-Time PCR with Serum Samples Is Indispensable for Early Diagnosis of Acute Q Fever

2009 ◽  
Vol 17 (2) ◽  
pp. 286-290 ◽  
Author(s):  
Peter M. Schneeberger ◽  
Mirjam H. A. Hermans ◽  
Erik J. van Hannen ◽  
Jeroen J. A. Schellekens ◽  
Alexander C. A. P. Leenders ◽  
...  
Keyword(s):  
Early Diagnosis ◽  
Phase I ◽  
Real Time ◽  
Acute Phase ◽  
Phase Ii ◽  
Real Time Pcr ◽  
Q Fever ◽  
Serum Samples ◽  
Convalescent Phase ◽  
Acute Q Fever

ABSTRACT The world's largest Q fever outbreak is ongoing in The Netherlands with around 3,000 confirmed cases since the first half of 2007. Increased awareness has resulted in early referral of patients for diagnostics. An important drawback to serological diagnosis of acute Q fever is the lag phase in antibody response. Therefore, we evaluated the performance of a real-time PCR for detection of Coxiella burnetii DNA using serum samples from patients with acute Q fever. PCR, targeting IS1111, was retrospectively performed on acute-phase and follow-up convalescent-phase serum samples from 65 patients with acute Q fever as diagnosed by immunofluorescence assay. The results obtained by PCR were related to disease stage as defined by subsequent appearance of phase II IgM, phase II IgG, phase I IgM, and phase I IgG (IgM-II, IgG-II, IgM-I, and IgG-I, respectively) antibodies and time since onset of disease. In addition, we analyzed seronegative acute-phase serum samples from patients with inconclusive Q fever serology, because no convalescent-phase serum samples were available. PCR was scored positive in 49/50 (98%) seronegative sera, 9/10 (90%) sera with isolated IgM-II antibodies, 3/13 (23%) sera with IgM-II/IgG-II antibodies, 2/41 (5%) sera with IgM-II/IgG-II/IgM-I antibodies, 0/15 (0%) sera with IgM-II/IgG-II/IgM-I/IgG-I antibodies, and 0/1 (0%) serum sample with IgM-II/IgG-II/IgG-I antibodies. The latest time point after onset of disease in which C. burnetii DNA could be detected was at day 17. In patients with inconclusive Q fever serology, PCR was positive in 5/50 (10%) cases. We conclude that real-time PCR with serum samples is indispensable for early diagnosis of acute Q fever. C. burnetii DNA becomes undetectable in serum as the serological response develops.

scholarly journals Evaluation of Two Rapid Immunochromatographic Assays for Diagnosis of Dengue among Vietnamese Febrile Patients

2007 ◽  
Vol 14 (6) ◽  
pp. 799-801 ◽  
Author(s):  
Tran Thi Thanh Nga ◽  
Khoa T. D. Thai ◽  
Hoang Lan Phuong ◽  
Phan Trong Giao ◽  
Le Quoc Hung ◽  
...  
Keyword(s):  
Sensitivity And Specificity ◽  
Acute Phase ◽  
Immunoglobulin G ◽  
Immunoglobulin M ◽  
Serum Samples ◽  
Rapid Tests ◽  
Immunosorbent Assays ◽  
Strip Test ◽  
Acute Phase Serum

ABSTRACT Results from two dengue rapid tests, the PanBio Duo cassette and the SD Bioline strip test, were compared to those of enzyme-linked immunosorbent assays (Focus Diagnostics) from sera of 200 Vietnamese febrile patients. The PanBio assay was superior, with sensitivity and specificity values for acute-phase serum samples of 54% and 70% (immunoglobulin M) and 70% and 88% (immunoglobulin G), respectively.

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