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

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.

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The value of repeat patient testing for SARS-CoV-2: real-world experience during the first wave

Access Microbiology ◽

10.1099/acmi.0.000239 ◽

2021 ◽

Vol 3 (7) ◽

Author(s):

Alex Zhu ◽

Margaret Creagh ◽

Chao Qi ◽

Shannon Galvin ◽

Maureen Bolon ◽

...

Keyword(s):

False Negative ◽

Laboratory Diagnosis ◽

Nucleic Acid Amplification ◽

Repeat Testing ◽

The Third ◽

Reverse Transcription Pcr ◽

Negative Results ◽

False Negative Results ◽

Number Of Patients ◽

Control And Prevention

Introduction. Reports of false-negative quantitative reverse transcription PCR (RT-qPCR) results from patients with high clinical suspension for coronavirus disease 2019 (COVID-19), suggested that a negative result produced by a nucleic acid amplification assays (NAAs) did not always exclude the possibility of COVID-19 infection. Repeat testing has been used by clinicians as a strategy in an to attempt to improve laboratory diagnosis of COVID-19 and overcome false-negative results in particular. Aim. To investigate whether repeat testing is helpful for overcoming false-negative results. Methods. We retrospectively reviewed our experience with severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) testing, focusing on the yield of repeat patient testing for improving SARS-CoV-2 detection by NAA. Results. We found that the yield from using repeat testing to identify false-negative patients was low. When the first test produced a negative result, only 6 % of patients tested positive by the second test. The yield decreased to 1.7 and then 0 % after the third and fourth tests, respectively. When comparing the results produced by three assays, the Centers for Disease Control and Prevention (CDC) SARS CoV-2 RT-qPCR panel, Xpert Xpress CoV-2 and ID NOW COVID-19, the ID NOW assay was associated with the highest number of patients who tested negative initially but positive on repeat testing. The CDC SARS CoV-2 RT-qPCR panel produced the highest number of indeterminate results. Repeat testing resolved more than 90 % of indeterminate/invalid results. Conclusions. The yield from using repeat testing to identify false-negative patients was low. Repeat testing was best used for resolving indeterminate/invalid results.

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Laboratory Diagnosis of African Horse Sickness: Comparison of Serological Techniques and Evaluation of Storage Methods of Samples for Virus Isolation

Journal of Veterinary Diagnostic Investigation ◽

10.1177/104063879000200108 ◽

1990 ◽

Vol 2 (1) ◽

pp. 44-50 ◽

Cited By ~ 34

Author(s):

Carol House ◽

Peter E. Mikiciuk ◽

Mary Lou Beminger

Keyword(s):

Virus Isolation ◽

Fluorescent Antibody ◽

False Negative ◽

Laboratory Diagnosis ◽

Enzyme Linked Immunosorbent Assay ◽

African Horse Sickness ◽

Serum Samples ◽

Negative Results ◽

Field Samples ◽

Storage Methods

Five serological methods of diagnosing African horse sickness were evaluated, using a battery of serum samples from experimental horses vaccinated and challenged with each serotype of African horse sickness virus (AHSV1 through AHSV9): agar gel immunodiffusion (AGID), indirect fluorescent antibody (IFA), complement fixation (CF), virus neutralization (VN), and enzyme-linked immunosorbent assay (ELISA). The 5 tests were also compared using a panel of field samples, convalescent equine sera with antibodies to domestic equine viral diseases, and sera from horses awaiting export. The ELISA described in this paper was group specific. It did not require calibration with a standard positive serum but did yield elevated values with negative sera that were repeatedly frozen and thawed or heat inactivated. The IFA test was sensitive but could not be used on some field sera as the control cells exhibited fluorescence, possibly due to the animal being recently vaccinated with cell culture material. Sixty-two experimental sera were compared by VN, CF, AGID, and ELISA. Forty sera, 10 positive and 30 negative, were correctly classified by the 5 serologic assays. The 22 remaining sera gave mixed reactions. The AGID had no false positive results but had false negative results for up to 20% of the samples, depending upon the comparison. The VN, CF, and ELISA were similar in their variability. The length of time that virus could be recovered from a viremic blood sample was compared in an evaluation of storage methods for virus isolation samples. Washed erythrocytes were held at 4 C, washed erythrocytes plus stabilizer were held at −70 C, and blood that was drawn into a preservative (oxalate/phenol/glycerol) was held at 4 C. Virus was isolated for 12 months from a sample stored as washed erythrocytes at 4 C but for only 6 months from the same blood stored by the other 2 methods.

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Development of an Enzyme-Linked Immunosorbent Assay for Immunoglobulin M Antibodies against Measles Virus

Clinical and Diagnostic Laboratory Immunology ◽

10.1128/cdli.10.3.439-442.2003 ◽

2003 ◽

Vol 10 (3) ◽

pp. 439-442 ◽

Cited By ~ 3

Author(s):

F. Roodbari ◽

M. H. Roustai ◽

A. Mostafaie ◽

H. Soleimanjdahi ◽

R. Sarrami Foroshani ◽

...

Keyword(s):

Immunosorbent Assay ◽

Neutralizing Antibodies ◽

Measles Virus ◽

Clinical Symptoms ◽

Maculopapular Rash ◽

Immunoglobulin M ◽

Enzyme Linked Immunosorbent Assay ◽

Serum Samples ◽

Vaccination Programs ◽

Elisa System

ABSTRACT Measles is a highly contagious respiratory virus infection, with typical clinical symptoms including maculopapular rash, fever, cough, coryza, and conjunctivitis. Despite implementation of widespread vaccination programs throughout the world, the rates of global morbidity and mortality are still considerable. This study was performed to design a reliable indirect enzyme-linked immunosorbent assay (ELISA) to measure measles-specific immunoglobulin M (IgM). First, human IgM was purified, and then an anti-IgM antibody was produced in rabbits and purified in a multistep process. The rabbit IgG against human IgM was conjugated with peroxidase. Measles virus-infected Vero cells produced viral antigen. One hundred serum samples from infants of 9 to 18 months of age, mostly vaccinated, were evaluated for determining the presence of specific IgM antibodies against measles virus. The samples were also evaluated for neutralizing antibodies against measles virus by a microneutralization test (MNT). By comparing the results of the ELISA with those of MNT, it was demonstrated that ELISA had a sensitivity and specificity of 100 and 92%, respectively. On the other hand, when the results obtained by our ELISA system were compared with those of an imported measles virus IgM ELISA kit (EIAgen; Adaltis Italia SPa, Bologna, Italy), a high level of agreement was shown (k = 0.926).

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False negative results in the Roche assay for HDL-cholesterol

Annals of Clinical Biochemistry International Journal of Laboratory Medicine ◽

10.1258/000456303322326533 ◽

2003 ◽

Vol 40 (5) ◽

pp. 572-575 ◽

Cited By ~ 6

Author(s):

Andries J Bakker ◽

Appie Zijlstra ◽

Marten P Leemhuis

Keyword(s):

Cardiovascular Disease ◽

Risk Evaluation ◽

False Negative ◽

Hdl Cholesterol ◽

Daily Practice ◽

Vascular Risk ◽

Serum Samples ◽

Negative Results ◽

Risk Estimates ◽

False Negative Results

Measurement of HDL-cholesterol is important in the risk evaluation of cardiovascular disease. Recently, we encountered a 61-year-old man with a negative result for HDL-cholesterol using the Roche assay. Further analysis revealed that this patient had a monoclonal Ig type IgM κ, with an electrophoresis-based concentration of 55.8 g/L, which apparently caused the negative result for HDL-cholesterol. Analysis of serum samples from other patients with monoclonal or polyclonal hypergammaglobulinaemia revealed that false negative results in the Roche assay for HDL-cholesterol may occur incidentally. Since negative results are detected easily in the laboratory, such patients do not pose a problem in daily practice. However, it is reasonable to suppose that false low results could also be caused by monoclonal as well as polyclonal Igs. Since such false low results remain undetected, vascular risk estimates consequently are altered and unnecessary treatment might be started.

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Five days of fever and myocardial inflammation

SAGE Open Medical Case Reports ◽

10.1177/2050313x211050891 ◽

2021 ◽

Vol 9 ◽

pp. 2050313X2110508

Author(s):

Keli D Coleman ◽

Paul Benz ◽

Nirzar S Parikh ◽

Danny G Thomas ◽

David Segar ◽

...

Keyword(s):

False Negative ◽

Clinical Signs ◽

Myocardial Inflammation ◽

Negative Results ◽

Pediatric Illness ◽

Multisystem Involvement ◽

False Negative Results ◽

Control And Prevention ◽

Evidence Based Guidelines ◽

Inflammatory Syndrome

Multisystem inflammatory syndrome in children is an emerging pediatric illness associated with severe acute respiratory syndrome coronavirus 2 infection. The syndrome is rare, and evidence-based guidelines are lacking. This report reviews a patient who presented for medical care multiple times early in the course of his illness, thus offering near-daily documentation of symptoms and laboratory abnormalities. The patient did not have thrombocytopenia, anemia, or myocardial inflammation until the fifth day of fever. These laboratory abnormalities coincided with the onset of rash, conjunctival injection, vomiting, and diarrhea: clinical signs that could serve as indicators for when to obtain blood tests. The timing of this patient’s onset of multisystem involvement suggests that testing for multisystem inflammatory syndrome in children after only 24 h of fever, as the Centers for Disease Control and Prevention recommends, may yield false-negative results. Testing for multisystem inflammatory syndrome in children after 4 days of fever may be more reliable.

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COVID-19 clinical and laboratory diagnosis overview

Journal of Egyptian Public Health Association ◽

10.1186/s42506-021-00087-w ◽

2021 ◽

Vol 96 (1) ◽

Author(s):

Rania A. Zayed ◽

Dalia Omran ◽

Abeer A. Zayed

Keyword(s):

False Negative ◽

Clinical Decision ◽

Laboratory Diagnosis ◽

Clinical Suspicion ◽

Laboratory Diagnostics ◽

Main Body ◽

Rt Pcr ◽

Serological Tests ◽

Negative Results ◽

Important Approach

Abstract Background COVID-19 was identified in Wuhan, China, in December 2019, and rapidly spread worldwide, being declared global pandemic on the 11th of March 2020. Since its emergence, COVID-19 has raised global concerns associated with drastic measures that were never adopted in any previous outbreak, to contain the situation as early as possible. Main body The 2019 novel corona virus (2019-nCoV) or SARS-CoV-2 is the causative agent of COVID-19. 2019-nCoV genetic sequence was rapidly identified within few days since the first reported cases and RT-PCR kits became available for COVID-19 diagnosis. However, RT-PCR diagnosis carries a risk of false-negative results; therefore, additional serologic tests are needed. In this review, we summarize the clinical scenario that raises suspicion of COVID-19 and available laboratory diagnostics. Conclusion The most important approach in the battle against COVID-19 is rapid diagnosis of suspicious cases, timely therapeutic intervention and isolation to avoid community spread. Diagnosis depends mainly on PCR testing and serological tests. However, even in the context of negative lab test results and clinical suspicion of COVID-19 infection, clinical decision should be based on clinical suspicion.

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

Journal of Clinical Microbiology ◽

10.1128/jcm.37.3.649-652.1999 ◽

1999 ◽

Vol 37 (3) ◽

pp. 649-652 ◽

Cited By ~ 26

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.

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Measles outbreak in a migrant laborer population in the wake of COVID-19 pandemic: Kathmandu, Nepal, 2020

10.21203/rs.3.rs-36529/v1 ◽

2020 ◽

Author(s):

Biplov Adhikari ◽

Sauharda Singh Karki ◽

Amrit Devkota ◽

Rahul Thakur ◽

Nabina Karki ◽

...

Keyword(s):

Measles Virus ◽

Health Centre ◽

Immunoglobulin M ◽

Enzyme Linked Immunosorbent Assay ◽

Serum Samples ◽

Measles Elimination ◽

Migrant Laborers ◽

Measles Outbreak ◽

Laboratory Confirmation ◽

Significant Difference

Abstract We describe a measles outbreak in migrant laborers working in a carpet factory in Kathmandu, the capital of Nepal. The outbreak occurred during the nationwide lockdown enforced to contain the COVID-19 pandemic and 2 weeks after the Supplmentary Immunization Activities done for measles elimination. We included all the patients from the factory presenting to Mulpani Primary Health Centre, Kathmandu, Nepal with acute febrile rash illness. Recovered patients with history of fever and rash within the defined outbreak period with a clear epidemiological linkage were also included. Laboratory confirmation was done by detection of immunoglobulin M (IgM) in serum samples against the measles virus via enzyme-linked immunosorbent assay (ELISA). We compared attack rates (ARs) between those less than 5 years and 5 years or older. We identified 11 case patients ranging from 7 months to 27 years of age (3 below 5years of age); rash onsets were from 28 March-20 April 2020. All case-patients were laborers who had immigrated to Kathmandu from rural parts of the country. We sent serum samples of 7 patients for laboratory confirmation; 5 patients tested positive. The remaining four patients had a clear epidemiological linkage. The average attack rate was 30.5% with no significant difference between attack rates among the <5 years and the ≥5 years reported as 37.5 and 28.5 respectively. Migrant population from rural regions with poor outreach of essential health services like vaccination can act as pockets of measles susceptible individuals if not measles reservoirs. The squalid conditions in which the migrant laborers live-in can also compound the risk of outbreak in such populations. It is prudent to address the vaccination status of such population and timely correct their vaccine status if unvaccinated in order to meet the goal of measles elimination by 2023.

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Evaluating polymicrobial immune responses in patients suffering from tick-borne diseases

Scientific Reports ◽

10.1038/s41598-018-34393-9 ◽

2018 ◽

Vol 8 (1) ◽

Cited By ~ 6

Author(s):

Kunal Garg ◽

Leena Meriläinen ◽

Ole Franz ◽

Heidi Pirttinen ◽

Marco Quevedo-Diaz ◽

...

Keyword(s):

Lyme Disease ◽

Immune Responses ◽

Opportunistic Infections ◽

Immunoglobulin M ◽

Enzyme Linked Immunosorbent Assay ◽

Serum Samples ◽

Microbial Infections ◽

Patient Will ◽

Control And Prevention ◽

The One

AbstractThere is insufficient evidence to support screening of various tick-borne diseases (TBD) related microbes alongside Borrelia in patients suffering from TBD. To evaluate the involvement of multiple microbial immune responses in patients experiencing TBD we utilized enzyme-linked immunosorbent assay. Four hundred and thirty-two human serum samples organized into seven categories followed Centers for Disease Control and Prevention two-tier Lyme disease (LD) diagnosis guidelines and Infectious Disease Society of America guidelines for post-treatment Lyme disease syndrome. All patient categories were tested for their immunoglobulin M (IgM) and G (IgG) responses against 20 microbes associated with TBD. Our findings recognize that microbial infections in patients suffering from TBDs do not follow the one microbe, one disease Germ Theory as 65% of the TBD patients produce immune responses to various microbes. We have established a causal association between TBD patients and TBD associated co-infections and essential opportunistic microbes following Bradford Hill’s criteria. This study indicated an 85% probability that a randomly selected TBD patient will respond to Borrelia and other related TBD microbes rather than to Borrelia alone. A paradigm shift is required in current healthcare policies to diagnose TBD so that patients can get tested and treated even for opportunistic infections.

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Three highly sensitive "bedside" serum and urine tests for pregnancy compared

Clinical Chemistry ◽

10.1093/clinchem/36.9.1686 ◽

1990 ◽

Vol 36 (9) ◽

pp. 1686-1688 ◽

Cited By ~ 10

Author(s):

H Christensen ◽

H H Thyssen ◽

O Schebye ◽

A Berget

Keyword(s):

Paired Comparison ◽

False Negative ◽

Diagnostic Sensitivity ◽

Enzyme Linked Immunosorbent Assay ◽

Serum Samples ◽

Negative Results ◽

False Negative Results ◽

Significant Difference ◽

Comparison Of The Results ◽

Urine And Serum

Abstract We examined three enzyme-linked immunosorbent assay (ELISA) kits for human choriogonadotropin (hCG) (pregnancy tests) for use with urine and serum samples: the Tandem Icon II hCG Urine and Tandem Icon II hCG Serum, the NovoClone Target hCG Test, and the Abbott TestPacks hCG-urine and hCG-serum. Paired comparison of the results from each kit indicated that the NovoClone Target assay showed significantly lower diagnostic sensitivity (P less than 0.05) than did the Tandem Icon II or Abbott TestPack, both for urine and for serum samples. None of the products demonstrated any significant difference (P greater than 0.05) in diagnostic specificity, but the NovoClone Target kit showed several serious false-negative results with both urine and serum. Paired testing of urine kits vs serum kits also showed no significant differences (P greater than 0.05) in diagnostic sensitivity or specificity. We found the Abbott kits to be the most convenient to use and to read.

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