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

2003 ◽  
Vol 10 (3) ◽  
pp. 439-442 ◽  
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).

scholarly journals Serological and Virological Characterization of Clinically Diagnosed Cases of Measles in Suburban Khartoum

2000 ◽  
Vol 38 (3) ◽  
pp. 987-991 ◽  
Author(s):  
H. Sittana El Mubarak ◽  
Marco W. G. Van De Bildt ◽  
Omer A. Mustafa ◽  
Helma W. Vos ◽  
Maowia M. Mukhtar ◽  
...  
Keyword(s):  
Neutralizing Antibodies ◽  
Measles Virus ◽  
Virus Isolation ◽  
Clinical Symptoms ◽  
Early Stage ◽  
Maculopapular Rash ◽  
Laboratory Diagnosis ◽  
Immunoglobulin M ◽  
Rt Pcr ◽  
Clinical Criteria

Measles continues to be a major childhood disease in terms of global morbidity and mortality. In the main areas of its endemicity the only available means of diagnosis are based on clinical criteria: the presence of a maculopapular rash and fever accompanied by cough, coryza, and/or conjunctivitis. We have studied 38 clinically diagnosed cases of measles in Khartoum, Sudan, by means of serology, reverse transcriptase PCR (RT-PCR) on throat swabs and virus isolation from lymphocytes. On the basis of serology, 28 patients were diagnosed as having an acute measles virus (MV) infection, while in 10 cases the clinical symptoms proved to have other causes. It was shown that in cases with low serum immunoglobulin M (IgM) levels, an additional measurement of IgG or virus-neutralizing antibodies was necessary to discriminate between patients with an acute MV infection sampled during an early stage of the disease and patients who had experienced an MV infection in the more distant past. The serological laboratory diagnosis was validated by an MV-specific RT-PCR: for all confirmed measles cases tested a fragment of the correct size which hybridized with a third MV-specific primer could be amplified, while all serologically negative cases were also RT-PCR negative. MV could be isolated from 17 out of 23 of the serologically confirmed cases, demonstrating that virus isolation is less reliable as a diagnostic tool than serology or RT-PCR. This study stresses the urgent need for a rapid diagnostic field test for measles.

scholarly journals Evaluation of an Immunoglobulin M Capture Enzyme-Linked Immunosorbent Assay for Diagnosis of Orientia tsutsugamushi Infection

2003 ◽  
Vol 10 (3) ◽  
pp. 394-398 ◽  
Author(s):  
Won-Jong Jang ◽  
Myung-Suk Huh ◽  
Kyung-Hee Park ◽  
Myung-Sik Choi ◽  
Ik-Sang Kim
Keyword(s):  
Immunosorbent Assay ◽  
Scrub Typhus ◽  
Microtiter Plate ◽  
Immunoglobulin M ◽  
Enzyme Linked Immunosorbent Assay ◽  
Orientia Tsutsugamushi ◽  
Serum Samples ◽  
Capture Elisa ◽  
Igm Antibodies ◽  
Immunodominant Antigen

ABSTRACT To differentiate scrub typhus from other acute febrile diseases, a rapid and reliable serological diagnosis is important. We developed an immunoglobulin M (IgM) capture enzyme-linked immunosorbent assay (ELISA) for diagnosis of recent Orientia tsutsugamushi infections in humans. The 56-kDa major outer membrane protein of O. tsutsugamushi is well known as the most immunodominant antigen in scrub typhus. The test is based on the use of the biotinylated recombinant 56-kDa protein of O. tsutsugamushi Boryong, Bor56, which was expressed as a fusion protein with a maltose-binding protein in Escherichia coli. In the test, the serum IgM antibodies were captured by anti-human IgM antibodies coated onto a microtiter plate. The captured IgM antibodies were revealed through sequential addition of biotinylated Bor56 antigen and peroxidase-conjugated streptavidin to the plate. The IgM capture ELISA was compared with the immunofluorescence antibody assay (IFA) by testing 176 serum samples from patients with diagnosed cases of rickettsial disease and patients with other acute febrile diseases. Of the 81 IgG IFA-positive samples, 78 tested positive (sensitivity, 96.3%) and all 31 IgM IFA-positive samples tested positive (sensitivity, 100%) by the IgM capture ELISA. The specificity of the IgM capture ELISA was 99%, and 1 of the 95 IFA-negative samples was positive in the assay. These results strongly suggest that IgM capture ELISA using the recombinant Bor56 antigen is a reliable and detailed method for the detection of early O. tsutsugamushi infection.

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

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.

scholarly journals Peptide-Based OspC Enzyme-Linked Immunosorbent Assay for Serodiagnosis of Lyme Borreliosis

1998 ◽  
Vol 36 (12) ◽  
pp. 3474-3479 ◽  
Author(s):  
Marianne J. Mathiesen ◽  
Michael Christiansen ◽  
Klaus Hansen ◽  
Arne Holm ◽  
Eva Åsbrink ◽  
...  
Keyword(s):  
Lyme Borreliosis ◽  
Immunosorbent Assay ◽  
Erythema Migrans ◽  
Immunoglobulin M ◽  
Enzyme Linked Immunosorbent Assay ◽  
Amino Acid Residues ◽  
Serum Samples ◽  
Igg Antibody ◽  
Acrodermatitis Chronica Atrophicans ◽  
Increased Sensitivity

Sera from 210 patients with Lyme borreliosis (LB) were studied by an enzyme-linked immunosorbent assay (ELISA) based on a synthetic peptide (pepC10) comprising the C-terminal 10-amino-acid residues of OspC of Borrelia burgdorferi. We found that 36.3 and 45.0% of the serum samples from patients with erythema migrans (EM) and neuroborreliosis (NB), respectively, displayed immunoglobulin M (IgM) anti-pepC10 reactivities, while these samples rarely (≤8%) displayed IgG antibody reactivities. Sera from patients with acrodermatitis chronica atrophicans did not contain anti-pepC10 antibodies. The diagnostic performance of this newly developed peptide ELISA was compared with those of an ELISA based on the full-length recombinant OspC protein (rOspC) and a commercially available ELISA based on theB. burgdorferi flagellum (Fla). The sensitivity of the IgM pepC10 ELISA was slightly lower (P < 0.04) than that of the rOspC ELISA for EM patients (36.3 versus 43.8%), while there was no difference for NB patients (45.0 versus 48.0%). However, the optical density values obtained by the pepC10 ELISA were generally higher than those obtained by the rOspC ELISA, leading to a significantly better quantitative discrimination between seropositive patients with NB and controls (P < 0.008). The specificity of the pepC10 ELISA was similar to those of the rOspC ELISA and the Fla ELISA for relevant controls including patients with syphilis and mononucleosis. Although the overall diagnostic sensitivity of the Fla ELISA was superior, 8.8 and 12.0% of the EM and NB patients, respectively, were antibody positive only by the pepC10 ELISA. Thus, use of a diagnostic test for LB based on the detection of IgM antibodies to pepC10 and Fla has increased sensitivity for the diagnosis of early LB.

scholarly journals Evaluation of Hemagglutinin Protein-Specific Immunoglobulin M for Diagnosis of Measles by an Enzyme-Linked Immunosorbent Assay Based on Recombinant Protein Produced in a High-Efficiency Mammalian Expression System

1998 ◽  
Vol 36 (12) ◽  
pp. 3509-3513 ◽  
Author(s):  
Fabienne B. Bouche ◽  
Nicolaas H. C. Brons ◽  
Sophie Houard ◽  
Francois Schneider ◽  
Claude P. Muller
Keyword(s):  
Recombinant Protein ◽  
Immunosorbent Assay ◽  
Measles Virus ◽  
High Efficiency ◽  
Expression System ◽  
Immunoglobulin M ◽  
Initial Time ◽  
Enzyme Linked Immunosorbent Assay ◽  
Mammalian Expression ◽  
Specific Immunoglobulin

Recombinant hemagglutinin (H) of the measles virus (MV) expressed in a mammalian high-expression system based on the Semliki Forest virus replicon was used in an enzyme-linked immunosorbent assay (ELISA) for the detection of specific immunoglobulin M (IgM) and IgG in patients with acute-phase measles. One hundred twelve serum specimens from 70 patients with measles were analyzed. Case definition was based on a commercial IgM ELISA that utilizes MV-infected cells (MV-ELISA) (Enzygnost; Behring Diagnostics); the clinical criteria of the Centers for Disease Control and Prevention (Atlanta, Ga.); and/or the increase in hemagglutinin test titers, neutralization test titers, and levels of MV-specific IgG whenever paired sera were available. The initial time courses of the IgM signal after the onset of rash are similar in the H- and MV-ELISAs. On days 0 to 19, both ELISAs detected IgM in 67 of 68 (98.5%) sera. Average maximal levels of IgM seem to persist, however, about 10 days longer in the MV-ELISA (up to day 25) than in the H-ELISA (day 15). From days 20 to 29 and 30 to 59, the H-ELISA detected only 64.3 (9 of 14) and 19.2% (5 of 26), respectively, of sera that were IgM positive by MV-ELISA. At least up to day 30, the performance of the H-ELISA seemed to be similar to that reported for commercial ELISAs based on whole MV. Our results demonstrate that MV H-specific IgM can be used to diagnose most measles cases from a single serum specimen collected within 19 days after the onset of rash and that the recombinant protein used in this study is suitable for this purpose.

scholarly journals Development of an Immunoglobulin M (IgM) Capture Enzyme-Linked Immunosorbent Assay for Detection of Equine and Swine IgM Antibodies to Vesicular Stomatitis Virus

2001 ◽  
Vol 8 (3) ◽  
pp. 475-481 ◽  
Author(s):  
En-min Zhou ◽  
Jose Riva ◽  
Alfonso Clavijo
Keyword(s):  
Vesicular Stomatitis Virus ◽  
Immunosorbent Assay ◽  
Polyclonal Antibodies ◽  
Competitive Elisa ◽  
Immunoglobulin M ◽  
Enzyme Linked Immunosorbent Assay ◽  
Serum Samples ◽  
Igm Antibodies ◽  
Vesicular Stomatitis ◽  
Elisa Plate

ABSTRACT An immunoglobulin M (IgM) capture enzyme-linked immunosorbent assay (MC-ELISA) was developed for the detection of primary infection of vesicular stomatitis virus (VSV) in equine and swine sera. The test was based on the use of biotinylated sheep antibodies against equine or swine IgM molecules bound to a streptavidin-coated ELISA plate. The captured IgM antibodies were detected by application of antigens prepared from the New Jersey and the Indiana VSV serotypes (VSV-NJ and VSV-IN, respectively) and mouse polyclonal antibodies against VSV-NJ and VSV-IN. The MC-ELISA was compared to a competitive ELISA (C-ELISA) and the standard microtiter serum neutralization (MTSN) assay by testing serum samples from horses and pigs experimentally infected with VSV-NJ or VSV-IN. The MC-ELISA detected specific homologous IgM antibodies from equine and swine sera as early as 5 and 4 days postinfection (DPI), respectively, and as late as 35 DPI. The MTSN test also detected antibodies as early as 5 DPI and as late as 160 DPI. In a similar fashion, the C-ELISA detected antibodies from 6 to 7 DPI and as late as 160 DPI. These results demonstrated that the MC-ELISA is a useful test for serodiagnosis of primary VSV infection in horses and pigs.

scholarly journals Enzyme-Linked Immunosorbent Assay Using Recombinant OspC and the Internal 14-kDa Flagellin Fragment for Serodiagnosis of Early Lyme Disease

1998 ◽  
Vol 36 (4) ◽  
pp. 857-861 ◽  
Author(s):  
Sebastian Rauer ◽  
Nicole Spohn ◽  
Christiane Rasiah ◽  
Uwe Neubert ◽  
Arnold Vogt
Keyword(s):  
Lyme Disease ◽  
Immunosorbent Assay ◽  
Erythema Migrans ◽  
Cell Extract ◽  
Sensu Stricto ◽  
Immunoglobulin M ◽  
Enzyme Linked Immunosorbent Assay ◽  
Epstein Barr Virus Infection ◽  
Serum Samples ◽  
Antigen Elisa

The outer surface protein C (OspC) and the internal 14-kDa flagellin fragment of strain GeHo of Borrelia burgdorferisensu stricto were expressed as recombinant proteins inEscherichia coli and were purified for use in an immunoglobulin M (IgM) enzyme-linked immunosorbent assay (OspC–14-kDa antigen ELISA). No hint at disturbing protein-protein interferences, which might influence the availability of immunoreactive epitopes, was found when the recombinant antigens were combined in the ELISA. The recombinant OspC–14-kDa antigen ELISA was compared to a commercial IgM ELISA that used a detergent cell extract from Borrelia afzelii PKo as the antigen. According to the manufacturer’s information, the cell extract contains, in addition to other antigens, the following diagnostically relevant antigens: the 100-kDa (synonyms, 93- and 83-kDa antigens), 41-kDa, OspA, OspC, and 17-kDa antigens. The specificity was adjusted to 95% on the basis of data for 154 healthy controls. On testing of 104 serum samples from patients with erythema migrans (EM), the sensitivity of the recombinant ELISA (46%) for IgM antibodies was similar to that of the commercial ELISA (45%). However, when 42 serum samples from patients with polyclonal B-cell stimulation due to an Epstein-Barr virus infection were tested, false-positive reactions were significantly less frequent in the recombinant ELISA (10%) than in the whole-cell-extract ELISA (23%). OspC displays sequence heterogeneity of up to 40% according to the genomospecies. However, when the reactions of serum specimens from controls and EM patients with OspC from representative strains of B. burgdorferi sensu stricto (strain GeHo) and B. afzelii (strain PKo) were compared in an ELISA, almost no differences in specificity and sensitivity were seen. This demonstrates that the sera predominantly recognize the common epitopes of OspC tested in this study. In conclusion, we suggest that the OspC–14-kDa antigens ELISA is a suitable test for the detection of an IgM response in early Lyme disease.

scholarly journals Detection of anti-avian bornavirus antibodies in parrots in the Czech Republic and Slovakia

2014 ◽  
Vol 83 (3) ◽  
pp. 195-199
Author(s):  
Martina Vondráčková ◽  
Viktor Tukač ◽  
Veronika Grymová ◽  
Pavlína Hájková ◽  
Zdeněk Knotek ◽  
...  
Keyword(s):  
Czech Republic ◽  
Immunosorbent Assay ◽  
Clinical Symptoms ◽  
Recent History ◽  
Enzyme Linked Immunosorbent Assay ◽  
Serum Samples ◽  
The Czech Republic ◽  
Avian Bornavirus ◽  
History Of ◽  
Proventricular Dilatation Disease

Since the avian bornavirus (ABV) aetiology of the proventricular dilatation disease (PDD) was proven in 2008, ABV has been detected in many avian species. The aim of the present study was to detect ABV antibodies in parrots in the Czech Republic and Slovakia. A total of 142 birds were examined, including 37 birds with symptoms typical for PDD, 54 birds without PDD symptoms, and 51 parrots without any clinical symptoms of PDD but originating from one flock with a proven history of PDD. Sera from 142 birds were tested using the enzyme-linked immunosorbent assay (ELISA) for detection of antibodies against ABV nucleoprotein p40. Of 142 serum samples, 71 were positive (50%) and 71 negative (50%). In a group of birds with clinical symptoms of PDD, 77.1% showed to be ABV positive, whereas in the group of sick birds without suspicion of PDD the percentage of positive birds was 31.6%. In the birds that had a cage mate that was positive for ABV or died with PDD, 42.9% were ABV positive. Of the parrots without PDD symptoms but originating from the flock with a recent history of PDD, 62.8% of the birds were positive for antibodies against ABV nucleoprotein p40. The results suggest that PDD is common and there is a high percentage of asymptomatic carriers of ABV in the breeding facilities of parrots in the Czech Republic and Slovakia.

scholarly journals Comparison of a Commercial Enzyme-Linked Immunosorbent Assay with Immunofluorescence and Complement Fixation Tests for Detection of Coxiella burnetii (Q Fever) Immunoglobulin M

2000 ◽  
Vol 38 (4) ◽  
pp. 1645-1647 ◽  
Author(s):  
Peter R. Field ◽  
Jody L. Mitchell ◽  
Avelina Santiago ◽  
David J. Dickeson ◽  
Sau-Wan Chan ◽  
...  
Keyword(s):  
Immunosorbent Assay ◽  
Coxiella Burnetii ◽  
Complement Fixation ◽  
Q Fever ◽  
Fluorescent Antibody ◽  
Reference Method ◽  
Immunoglobulin M ◽  
Enzyme Linked Immunosorbent Assay ◽  
Serum Samples ◽  
Igm Elisa

A commercially available enzyme-linked immunosorbent assay (ELISA) for the diagnosis of Q fever (PanBio Coxiella burnetiiimmunoglobulin M [IgM] ELISA, QFM-200) was compared to the indirect fluorescent antibody test (IFAT) for C. burnetii IgM and the complement fixation test (CFT). The ELISA demonstrated 92% agreement with the reference method (IFAT), and gave a sensitivity of 99% (69 of 70 samples) and a specificity of 88% (106 of 121). Specificity can be increased with confirmation by IFAT. CFT was found to have a specificity of 90% (107 of 119), although it was lacking in sensitivity (73%; 51 of 70). No cross-reactivity was observed in the ELISA with serum samples from patients with mycoplasma (n = 6), chlamydia (n = 5), or legionella (n = 4) infections, although 2 of 5 patients with leptospirosis and 1 of 4 samples containing rheumatoid factor (RF) demonstrated positive results in the ELISA. Results indicate that the performance of the PanBio C. burnetii (Q fever) IgM ELISA (F = 187) is superior to that of CFT (F = 163), and consequently the ELISA should be a useful aid in the diagnosis of acute Q fever.

scholarly journals Diagnosis of Chikungunya Fever in an Indian Population by an Indirect Enzyme-Linked Immunosorbent Assay Protocol Based on an Antigen Detection Assay: a Prospective Cohort Study

2009 ◽  
Vol 17 (2) ◽  
pp. 291-297 ◽  
Author(s):  
Rajpal Singh Kashyap ◽  
Shweta H. Morey ◽  
Sonali S. Ramteke ◽  
Nitin H. Chandak ◽  
Manmohan Parida ◽  
...  
Keyword(s):  
Immunosorbent Assay ◽  
Disease Process ◽  
Acute Stage ◽  
Antigen Detection ◽  
Immunoglobulin M ◽  
Enzyme Linked Immunosorbent Assay ◽  
Serum Samples ◽  
Chikv Infection ◽  
Process Diagnosis ◽  
Nagpur District

ABSTRACT A Chikungunya virus (CHIKV) outbreak continues in India. Monitoring of the clinical features of CHIKV infection is an important component of assessing the disease process. Diagnosis is usually made by an immunoglobulin M (IgM)/IgG enzyme-linked immunosorbent assay (ELISA). However, these assays have extremely low sensitivities for the detection of infection in the majority of CHIKV patients during the acute stage of infection (during the 1 to 4 days after infection). In our laboratory, a sensitive ELISA protocol for antigen detection has been developed for the detection of CHIKV infection in the acute stage, and in the present study we assessed the usefulness of this ELISA-based system for the detection of CHIKV infection. We performed a prospective, double-blinded study of 205 Indian patients with suspected CHIKV infection in the Nagpur District. All patients underwent a full clinical assessment, and their serum samples were analyzed for the presence of antigens and of IgM and IgG by an ELISA protocol. In patients with CHIKV infection, the sensitivity of antigen detection was 85%, which was significantly higher (P < 0.001) than that of IgM (17%) or IgG (45%) detection. The sensitivity of IgM (20%) or IgG (25%) detection was significantly lower than that of the antigen assay (95%) for patients with acute infections (i.e., from day 1 to day 5 after infection). Antigen detection not only gives a positive confirmatory result in the early phase of the disease, but it is also useful in the prodromal and subclinical stage and may be useful for field applications for the rapid detection of CHIKV infection.

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