scholarly journals Analysis of Complement Fixation and Commercial Enzyme Immunoassays for Detection of Antibodies to Mycoplasma pneumoniae in Human Serum

2000 ◽  
Vol 7 (5) ◽  
pp. 778-780 ◽  
Author(s):  
W. Lanier Thacker ◽  
Deborah F. Talkington
Keyword(s):  
Mycoplasma Pneumoniae ◽  
Complement Fixation ◽  
Adult Population ◽  
Immunoglobulin M ◽  
Igg Antibodies ◽  
Serum Samples ◽  
Enzyme Immunoassays ◽  
Current Infection ◽  
Paired Serum ◽  
Antibody Tests

ABSTRACT The Meridian ImmunoCard (IC), GenBio ImmunoWELL-IgM, and Remel EIA commercial antibody tests are qualitative enzyme immunoassays that detect antibodies to Mycoplasma pneumoniae in serum. These tests were compared to an M. pneumoniae complement fixation (CF) assay, which uses a commercially available antigen component. The Meridian IC and the ImmunoWELL-IgM detect immunoglobulin M (IgM) only; the Remel EIA and the CF test detect both IgM and IgG antibodies. Detection of specific IgM antibody, which appears early in infection, can be, but is not always, indicative of a recent or current infection. Paired serum samples from 64 adult patients with probable M. pneumoniae infection were examined with each of the four tests. Thirty (47%) of the 64 acute-phase sera were IgM positive by Meridian IC, 26 (41%) were positive by Remel EIA, 24 (38%) were positive by CF, and 15 (23%) were positive by ImmunoWELL-IgM. When both the acute- and convalescent-phase serum samples from each patient were examined, 61 (95%) of the 64 patients were positive by CF, 60 patients (94%) were positive by Remel EIA, 52 patients (81%) were IgM positive by the Meridian IC, and 29 patients (45%) were IgM positive by the ImmunoWELL-IgM assay. The Meridian IC was more sensitive than the other tests for early detection of IgM antibodies. However, after examining paired serum samples, we concluded that the detection of IgM alone may not be useful for all cases of mycoplasma infection, especially in an adult population.

The Implication of Viruses in Idiopathic Sudden Hearing Loss: Primary Infection or Reactivation of Latent Viruses?

1981 ◽  
Vol 89 (1) ◽  
pp. 137-141 ◽  
Author(s):  
Robert W. Veltri ◽  
William R. Wilson ◽  
Philip M. Sprinkle ◽  
Susan M. Rodman ◽  
Debra A. Kavesh
Keyword(s):  
Hearing Loss ◽  
Influenza Virus ◽  
Herpes Simplex ◽  
Mycoplasma Pneumoniae ◽  
Primary Infection ◽  
Sudden Hearing Loss ◽  
Serum Samples ◽  
Paired Serum ◽  
Group B

Seventy-seven paired serum samples from patients with known idiopathic sudden hearing loss (ISHL) were surveyed using viral serologic methods. Fifteen different viruses and Mycoplasma pneumoniae were the agents tested. We determined an incidence of 65% (49/77) of documented significant seroconversions to one or more of the agents surveyed. Multiple agents were involved in 24 of the 49 positive cases we studied. Influenza virus Group B in 14 (18%) and rubeola in 12 (16%) were the most prevalent, followed by Herpes simplex type 1 in 6 (8%), mumps in 6 (8%), influenza Group A3 in 6 (8%), rubella in 5 (7%), and cytomegalovirus (CMV) in 5 (7%).

scholarly journals Detection of antibodies to decorin-binding protein A (DbpA) and DbpB after infection of dogs with Borrelia burgdorferi by tick challenge

2020 ◽  
Vol 32 (3) ◽  
pp. 481-485
Author(s):  
Darby G. Oldenburg ◽  
Dean A. Jobe ◽  
Steven D. Lovrich ◽  
Rhonda L. LaFleur ◽  
Douglas W. White ◽  
...  
Keyword(s):  
Lyme Disease ◽  
Borrelia Burgdorferi ◽  
Ixodes Scapularis ◽  
Binding Protein ◽  
Protein A ◽  
Immune Serum ◽  
Immunoglobulin M ◽  
Antibody Responses ◽  
Igg Antibodies ◽  
Serum Samples

We characterized the antibody response to decorin-binding protein A (DbpA) or DbpB from immune serum samples collected from 27 dogs infected with Borrelia burgdorferi by Ixodes scapularis ticks. Immunoglobulin M (IgM) antibodies to DbpA or DbpB were rarely detected, but high levels of IgG antibodies to DbpA were detected in 16 of 27 of the immune sera collected 1 mo after infection, 20 of 25 of the sera collected after 2 mo, and each of the 23, 17, or 11 serum samples evaluated after 3, 4, or 5 mo, respectively. In addition, IgG antibodies to DbpB were detected in 22 of 27 ( p = 0.005) tested dogs after 1 mo, and the frequency of detecting the antibodies thereafter closely mimicked the antibody responses to DbpA. Moreover, antibodies to DbpA or DbpB were not produced by dogs vaccinated with a whole-cell B. burgdorferi bacterin; removing the antibodies to DbpA by adsorption to recombinant DbpA (rDbpA) did not affect the reactivity detected by a rDbpB ELISA. Therefore, the findings from our preliminary study showed that antigenically distinct antibodies to DbpA or DbpB are produced reliably during canine infection with B. burgdorferi, and the response is not confounded by vaccination with a Lyme disease bacterin. Larger studies are warranted to more critically evaluate whether detecting the antibody responses can improve serodiagnostic confirmation of canine Lyme disease.

scholarly journals Evaluation of SARS-CoV-2 Antibody Point of Care Devices in the Laboratory and Clinical Setting

2021 ◽  
Author(s):  
Kirsty McCance ◽  
Helen Wise ◽  
Jennifer Simpson ◽  
Becky Bachelor ◽  
Harriet Hale ◽  
...  
Keyword(s):  
Immune Status ◽  
Point Of Care ◽  
Capillary Blood ◽  
Serum Samples ◽  
Specificity And Sensitivity ◽  
Independent Evaluation ◽  
Positive Antibody ◽  
Antibody Titres ◽  
Paired Serum ◽  
Antibody Tests

SARS-CoV-2 Antibody tests have been marketed to diagnose previous SARS-CoV-2 infection and as a test of immune status. There is a lack of evidence on the performance and clinical utility of these tests. We aimed to carry out an evaluation of 14 point of care (POC) SARS-CoV-2 antibody tests. Serum from participants with previous RT-PCR (Real-Time Polymerase chain reaction) confirmed SARS-CoV-2 infection and pre-pandemic controls were used to determine specificity and sensitivity of each POC device. Changes in sensitivity with increasing time from infection were determined on a cohort of participants. Corresponding neutralising antibody status was measured to establish whether the detection of antibodies by the POC device correlated with immune status. Paired capillary and serum samples were collected to ascertain whether POC devices performed comparably on capillary samples. Sensitivity and specificity varied between the POC devices and in general did not meet the manufacturers reported performance characteristics signifying the importance of independent evaluation of these tests. The sensitivity peaked at >20 days following symptoms onset however sensitivity of 3 POC devices evaluated at extended time points showed that sensitivity declined with time and this was particularly marked at >140 days post infection onset. This is relevant if the tests are to be used for sero-prevelence studies. Neutralising antibody data showed positive antibody results on POC devices did not necessarily confer high neutralising antibody titres and these POC devices cannot be used to determine immune status to the SARS-CoV-2 virus. Comparison of paired serum and capillary results showed that there was a decline in sensitivity using capillary blood. This has implications in the utility of the test as they are designed to be used on capillary blood by the general population.

Clinical utility of loop-mediated isothermal amplification for rapid diagnosis of Mycoplasma pneumoniae in children

2014 ◽  
Vol 63 (2) ◽  
pp. 248-251 ◽  
Author(s):  
Yuta Aizawa ◽  
Tomohiro Oishi ◽  
Shinya Tsukano ◽  
Tetsuo Taguchi ◽  
Akihiko Saitoh
Keyword(s):  
Mycoplasma Pneumoniae ◽  
Lamp Assay ◽  
Cost Effective ◽  
Community Acquired Pneumonia ◽  
Isothermal Amplification ◽  
City Hospital ◽  
Serum Samples ◽  
Loop Mediated Isothermal Amplification ◽  
Paired Serum ◽  
Median Interval

Loop-mediated isothermal amplification (LAMP) is a cost-effective and rapid method for identifying Mycoplasma pneumoniae (MP). We investigated the utility of the LAMP assay in diagnosing MP pneumonia among children in a clinical setting. In this prospective study, the cause of community-acquired pneumonia was evaluated in 111 patients for whom MP was the suspected pathogen. All participants were patients at a city hospital in Japan between April 2012 and September 2012. Throat swabs for the LAMP assay were obtained at admission, and paired serum samples to measure antibody titres to MP by particle agglutination were obtained at admission and during convalescence. Overall, 45 of 111 (41 %) patients had a fourfold or greater increase in MP titres and received a diagnosis of MP pneumonia. Among them, 43 (96 %) patients (median age, 9 years) were positive on the LAMP assay and had a fourfold or greater increase in MP titres. The median interval from fever onset to collection of throat swabs was 7 days (range, 4–10 days). As compared with paired serum titres, the LAMP assay enabled quicker diagnosis of MP (median interval, 13 vs. 7 days), thereby allowing early initiation of appropriate antimicrobial therapy.

scholarly journals Antibody responses toBorrelia burgdorferiouter surface proteins C and F in experimentally infected Beagle dogs

2015 ◽  
Vol 27 (4) ◽  
pp. 526-530 ◽  
Author(s):  
Steven M. Callister ◽  
Rhonda L. LaFleur ◽  
Dean A. Jobe ◽  
Steven D. Lovrich ◽  
Terri L. Wasmoen
Keyword(s):  
Ixodes Scapularis ◽  
Immunoglobulin M ◽  
Surface Proteins ◽  
Beagle Dogs ◽  
Serum Samples ◽  
Joint Tissue ◽  
Joint Infections ◽  
Outer Surface Proteins ◽  
Antibody Levels ◽  
Antibody Tests

Antibody levels to outer surface proteins C and F (OspC and OspF, respectively) in sera collected from laboratory Beagle dogs at 1, 2, and 4 months after challenge with infected black-legged ticks ( Ixodes scapularis) were determined. Each dog was confirmed by culture to harbor Borrelia burgdorferi in the skin ( n = 10) or the skin and joints ( n = 14). Significant levels of immunoglobulin M (Ig)M or IgG anti-OspC antibodies were detected in single serum samples from only 3 (13%) dogs. Similarly, IgM anti-OspF antibodies were detected in only 1 (4%) serum sample collected from a dog with B. burgdorferi in the skin and joints. In contrast, 4 (29%) dogs with skin and joint infections produced IgG anti-OspF antibodies after 2 months, and the response expanded to include 2 (20%) dogs with skin infection and 4 additional dogs with skin and joint infections (overall sensitivity = 62%) after 4 months. The findings failed to support the utility of OspC-based antibody tests for diagnosing canine Lyme disease, but demonstrated that dogs with B. burgdorferi colonizing joint tissue most often produced significant levels of IgG anti-OspF antibodies. Therefore, additional studies to more thoroughly evaluate the clinical utility of OspF-based antibody tests are warranted.

scholarly journals Role of rheumatoid factor in complement fixation and indirect hemagglutination tests for immunoglobulin M antibody to cytomegalovirus

1978 ◽  
Vol 8 (2) ◽  
pp. 160-165
Author(s):  
N E Cremer ◽  
M Hoffman ◽  
E H Lennette
Keyword(s):  
Rheumatoid Factor ◽  
Antibody Titer ◽  
Gamma Globulin ◽  
Complement Fixation ◽  
Immunoglobulin M ◽  
Serum Samples ◽  
Igg Antibody ◽  
Indirect Hemagglutination ◽  
Indirect Hemagglutination Test

Absorption of immunoglobulin M (IgM)-rheumatoid factor (RF) from serum samples by reaction with insolubilized gamma globulin reduced the complement-fixing (CF) antibody titer to cytomegalovirus (CMV) antigen to less than 1:2 in the IgM fraction of some, but not all, sera. Thus, IgM-CF activity in some sera appeared to be due to specific IgM anti-CMV antibody and in other sera to complexes of IgM-RF with antiviral IgG antibody. Prozones were present in the CF tests on IgM fractions. Increasing the concentration of antigen from 2 to 4 U reduced the prozone titer by one or two double dilutions. This observation suggested that a competition for antigen may be operating at low dilutions of IgM antibody fractions. Removal of RF had little or no effect on the reaction of the IgM fraction of sera with CMV by the indirect hemagglutination test.

scholarly journals SARS-CoV-2 Seroprevalence in a Cohort of Asymptomatic, RT-PCR Negative Croatian First League Football Players

2020 ◽  
Author(s):  
Adriana Vince ◽  
Renata Zadro ◽  
Zvonimir Šostar ◽  
Sunčanica Ljubin Sternak ◽  
Jasmina Vraneš ◽  
...  
Keyword(s):  
Football Players ◽  
Professional Football ◽  
Igg Antibodies ◽  
Rt Pcr ◽  
Antibody Assay ◽  
Serum Samples ◽  
Staff Members ◽  
Paired Serum ◽  
Serology Testing

AbstractBackgroundDuring the COVID-19 pandemic the Croatian Football Federation has launched a new model of pre-season systematic examination of football players, emphasizing the diagnosis of asymptomatic SARS-CoV-2 infection and preventing further spread among the players.ObjectivesThe aim of this study was to assess the prevalence and dynamics of SARS-CoV-2 IgA and IgG antibodies in the cohort of asymptomatic and SARS-CoV-2 PCR negative professional football players in the Croatian First Football League by using a commercial ELISA antibody assay in the paired serum samples taken 2 months apart.MethodsSerology testing was performed from May till July 2020 in a cohort of 305 asymptomatic football players and club staff members. RT-PCR for detection of SARS-CoV-2 from nasopharyngeal swabs was performed on three occasions, and Euroimmun ELISA for detection of IgA and IgG (S1 and NCP) antibodies was tested in paired serum samples in May and July.ResultsAll RT-PCR results were negative. Sixty-one (20%) participants were reactive in one or two classes of antibodies at baseline and/or follow-up serology testing. IgA reactivity was found in 41 (13.4% [95% CI=10.7-17.7]) baseline sera and 42 (13.8% [95% CI=10.3-18.9]) follow-up sera. IgG to S1 protein was found in 6 (2% [95% CI=0.9-4.2]) participants at baseline and 1 (0.33% [95% CI=0.0006-1.83]) at follow-up. IgG to NCP was found in 2 (0.7% [95% CI=0.2-2.4]) participants at baseline and 8 (2.6% (95% CI=1.3-5.1]) participants at follow-up. Noticeable dynamics in the paired sera was observed in 18 (5.9%) participants (excluding borderline IgA results) or 32 (10.5%) (including IgA borderline results).ConclusionVarious patterns of IgA and IgG reactivity were found in the paired serum samples. Based on serology dynamics we estimate that in 5.9%-10.5% of PCR negative football players asymptomatic exposure to SARS-CoV-2 during pandemics could not be excluded.

scholarly journals Serum Dried Samples to Detect Dengue Antibodies: A Field Study

2017 ◽  
Vol 2017 ◽  
pp. 1-6 ◽  
Author(s):  
Angelica Maldonado-Rodríguez ◽  
Othon Rojas-Montes ◽  
Guillermo Vazquez-Rosales ◽  
Adolfo Chavez-Negrete ◽  
Magdalena Rojas-Uribe ◽  
...  
Keyword(s):  
Field Study ◽  
Igg Antibodies ◽  
Specific Detection ◽  
Serum Samples ◽  
Paired Samples ◽  
Paired Serum

Background. Dried blood and serum samples are useful resources for detecting antiviral antibodies. The conditions for elution of the sample need to be optimized for each disease. Dengue is a widespread disease in Mexico which requires continuous surveillance. In this study, we standardized and validated a protocol for the specific detection of dengue antibodies from dried serum spots (DSSs).Methods. Paired serum and DSS samples from 66 suspected cases of dengue were collected in a clinic in Veracruz, Mexico. Samples were sent to our laboratory, where the conditions for optimal elution of DSSs were established. The presence of anti-dengue antibodies was determined in the paired samples.Results. DSS elution conditions were standardized as follows: 1 h at 4°C in 200 µl of DNase-, RNase-, and protease-free PBS (1x). The optimal volume of DSS eluate to be used in the IgG assay was 40 µl. Sensitivity of 94%, specificity of 93.3%, and kappa concordance of 0.87 were obtained when comparing the antidengue reactivity between DSSs and serum samples.Conclusion. DSS samples are useful for detecting anti-dengue IgG antibodies in the field.

A Prospective Study of the Persistence of Mycoplasma Pneumoniae Antibody Levels

1974 ◽  
Vol 19 (3) ◽  
pp. 129-133 ◽  
Author(s):  
G. R. Jones ◽  
Sheila M. Stewart
Keyword(s):  
Prospective Study ◽  
Mycoplasma Pneumoniae ◽  
Complement Fixation ◽  
Metabolic Inhibition ◽  
Inhibition Test ◽  
Cold Agglutinins ◽  
Current Infection ◽  
A Prospective Study ◽  
Antibody Levels ◽  
A Current

Sera from 13 patients with initial complement fixation (CF) titres of 160 or above, and from 5 with rising titres to Mycoplasma pneumoniae were tested at 2-monthly intervals for CF antibodies and for cold agglutinins. In an attempt to differentiate between ‘early’ and ‘late’ sera with raised titres, initial sera and those, taken 6 to 8 months later, with a CF titre of 160 or above were tested by the metabolic inhibition test and also treated with 2-mercaptoethanol (2ME). Cold agglutinins were detected in 5 of the 13 initial sera but in none of the 7 ‘late’ sera with raised CF titres. A 4-fold or greater fall in the CF titre after treatment with 2ME occurred in 5 of the 12 initial sera tested, and in one of the 7 ‘late’ sera. Metabolic inhibiting antibodies were present in 7 of the 12 initial sera and in 4 of the 6 ‘late’ sera tested. In only one patient with a raised initial CF titre but no cold agglutinins and no 2ME effect was there later evidence of a current infection with M. pneumoniae. Cold agglutinins were present and/or there was a 2ME effect in the initial sera from all 5 patients with rising titres. It is concluded that a combination of cold agglutinins and the effect of 2ME on the CF titres will help to differentiate ‘early’ from ‘late’ sera with significantly raised CF antibodies.

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.

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