IgM In Serum Samples Research Articles

Measles-Clinical and Biological Manifestations in Adult Patients, Including a Focus on the Hepatic Involvement: Results from a Single-Center Observational Cohort Study from Romania.

Background: Hepatic involvement in measles, particularly among adolescents and adults, has been recognized since 1960. This involvement typically manifests during the eruptive phase of the disease and is primarily characterized by hepatocellular dysfunction, with jaundice being a less common occurrence. Studies have reported hepatic involvement in 80-86% of measles cases among young adults associated with severe forms of the disease with intra-infectious hepatitis. Recent data from Romania indicated 20,035 confirmed measles cases between January and June 2024, including 17 fatalities and significant hepatic alterations. These findings underscore the need for the further investigation of the hepatic manifestations of measles. The primary objective of our study was to evaluate the clinical and baseline characteristics of the enrolled patients with a brief assessment of liver impairment. Methods: In light of these observations, we conducted a retrospective analysis between 1 November 2023 and 15 June 2024 in patients aged >16 years who were confirmed, by the detection of measles IgM in serum samples, to have acute measles infection and hospitalization. Results: During the study period, 71 hospitalized patients were diagnosed with measles, of whom 37 were female (52.1%), with ages ranging from 16 to 64 years (mean age 34.21 years). Most cases (77.5%) exhibited moderate clinical forms of measles, while 22.5% had severe forms. Respiratory failure requiring oxygen therapy was uncommon (25.4% of severe pneumonia cases). Although a Pearson chi-square test indicated no significant association between the presence of pneumonia and clinical form (p = 0.066), the likelihood ratio test suggested a potential link (p = 0.018). Hepatic involvement was common with elevated AST (87.3%) and ALT (76%) levels. Jaundice was observed in 12.7% of patients. GGT changes were noted in 35.2% of cases, with significant correlations between GGT levels and disease severity (p = 0.001). Analysis of various symptoms revealed significant associations between nausea, dyspnea, and severe clinical forms. Anorexia, diarrhea, and nausea were the most frequently reported symptoms. Thrombocytopenia was observed in 11 patients, with no significant correlation with disease severity. Comorbidities, such as COPD, were significantly associated with disease severity (p = 0.010). Conclusions: Our findings highlight that, while cytolytic hepatic damage is a typical response to measles infection, cholestatic involvement may serve as an indicator of more severe disease progression.

Comparative evaluation of chemiluminescent immunoassay and enzyme-linked immunosorbent assays for the diagnosis of West Nile virus infections.

In August 2020, anew West Nile virus (WNV) outbreak affected 71 people with meningoencephalitis in Andalusia (Spain). Samples from these individuals were received in our laboratory, a regional Virus Referral Centre. The aim of this study was to compare the agreement, sensitivity and specificity of findings between the WNV VIRCLIA IgG and IgM assay (Vircell, Spain) and the WNV ELISA IgM and IgG assay (Euroimmun, Germany) and to compare the performance of WNV VIRCLIA IgM and Euroimmun ELISA for cerebrospinal fluid (CSF) diagnosis. The study included 24 CSF samples (paired with serum samples) and 247 serum samples from 217 patients with suspected WNV infection (1 or 2 per patient). The agreement between ELISA and CLIA tests for IgM and Ig G detection in serum was 93% (kappa index = 0.85) and 96% (kappa index = 0.89) respectively. Sensitivity values of ELISA and CLIA tests for IgM in serum samples were 96.7% and 98.9%, respectively, and specificity values were 96.4% and 95.4% respectively. Sensitivity values of ELISA and CLIA test for IgG in serum samples were 91.1% and 97%, respectively, and specificity values were 100% and 98.8% respectively. Results obtained with ELISA and CLIA tests in CSF samples showed 75% agreement between them (kappa index = 0.51). According to these findings, the WNV VIRCLIA IgM and IgG monotest offers an accurate qualitative detection of WNV in serum and CSF specimens.

Differential Serodiagnostics of Latent Stages of Syphilis Based on Measuring IgG and IgM Levels towards Extended Panel of Recombinant Antigens of T. pallidum.

Immunochips containing 12 recombinant antigens of T. pallidum (Тр15, Тр17, Тр47, TmpA, Тр0163, Тр0277, Тр0319, Тр0453, Тр0684, Тр0965, Тр0971, and Тр1038) were prepared to assay for IgG and IgM in serum samples (n=68) of healthy individuals and patients with the latent stages of syphilis. The linear discriminant analysis of detected IgG and IgM differentiated three groups of serum samples as 1) early latent syphilis; 2) seroresistant early latent syphilis; and 3) late latent syphilis with overall differentiation potency of 95.6% (88.9-100%). The samples of all syphilis patients were differentiated from the samples of healthy individuals with 100% specificity.

Depletion of interfering IgG and IgM is critical to determine the role of IgE in pegvaliase-associated hypersensitivity

Pegvaliase is an enzyme substitution therapy developed to lower blood phenylalanine (Phe) in adults with phenylketonuria (PKU). In phase 3 clinical studies, pegvaliase substantially reduced mean blood Phe in adult subjects with PKU. The most common type of adverse event observed in the pegvaliase clinical program was hypersensitivity adverse events (HAEs), which included occurrences of arthralgia, rash, and pruritis. The most clinically relevant HAEs were acute systemic hypersensitivity reactions consistent with anaphylaxis observed in 4.6% of phase 3 patients. HAEs were more commonly observed around the time of high circulating immune complex (CIC) levels and complement activation, and the majority of subjects that experienced acute systemic hypersensitivity events were able to continue treatment, which is atypical for a classical IgE-mediated anaphylactic event, but common for type III hypersensitivity reactions.To investigate the alternative mechanism of type III hypersensitivity, serum samples collected shortly after hypersensitivity events (in phase 2 and 3 studies) were tested for anti-pegvaliase IgE using custom radioallergosorbent test and/or ImmunoCAP® (ThermoFisher Scientific, Waltham, MA) assay methods. All subjects with acute systemic hypersensitivity that were tested for anti-pegvaliase IgE at or near the time of event with one or both assays tested negative for IgE. As presented here, an investigation using selected study samples with high anti-drug antibody (ADA) titers demonstrated that presence of IgM and/or IgG ADA can interfere with measurement of a human anti-pegvaliase IgE surrogate positive control. A depletion method was therefore developed using protein A- and G-conjugated Sepharose to remove interfering IgG and IgM in serum samples to low levels (<45 mg/dL) before IgE testing. A final 2× concentration step brought the IgE concentration in the depleted sample to approximately the same level of the starting serum. Phase 3 study samples with sufficient volume remaining that previously tested negative for anti-pegvaliase IgE were re-tested after depletion of IgG and IgM. All samples again tested negative, confirming the original test results. Taken together, the clinical presentation, temporal association of HAEs with CIC levels and complement activation, and lack of anti-pegvaliase IgE suggest pegvaliase-associated acute systemic hypersensitivity events were not IgE-mediated. Furthermore, we describe a universal method that is broadly applicable to enzyme therapies for detection of low concentrations of drug-specific IgE in the presence of high titer anti-drug antibodies of different isotypes.

Serum anti-phenolic glycolipid-1 IgA correlates to IgM isotype in leprosy patients: a possible candidate for seroepidemiological surveys?

The aim of this study was to compare serum anti-phenolic glycolipid-1 IgA, IgG, and IgM levels in leprosy patients and controls. Analysis of anti-PGL-1 IgA, IgG, or IgM in serum samples from multibacillary (MB, n=32) and paucibacillary (PB, n=22) leprosy patients, and in non-endemic controls (n=17), using an indirect enzyme-linked immunosorbent assay. A strong correlation between serum IgM and IgA isotypes was found (r=.745, P<.0001) in MB patients. A moderate correlation was found in all analyses in PB patients. A moderate agreement was found between anti-PGL1 IgA and IgM tests. Based on the ROC curves, the cut-off values were selected and the parameters of validation were calculated. Considering the clinical forms altogether, the diagnostic sensitivities were 50.0% for IgA, 22.2% for IgG, and 74.1% for IgM. The positive (VPP) and negative (VPN) predictive values were estimated for each isotype. For IgA, the VPP and VPN were, respectively, 100.0% (87.0%-100.0%; 95% confidence interval) and 38.7% (24.4%-54.5%); for IgG, 100% (87.0%-100.0%) and 28.8% (17.8%-42.1%), respectively; and for IgM, 95.2% (83.8%-99.4%) and 51.7% (32.5%-70.6%), respectively. Despite the limiting factors, anti-PGL1 IgA correlates to IgM levels and it could be considered as a possible laboratorial tool to be also used, for instance, in serological follow-up studies.

Humoral immune responses of experimentally Eimeria ninakholyakimovae-infected goat kids.

Although cellular immune reactions seem to be crucial for protective immune responses in Eimeria spp. infections, there are also evidences on an active involvement of the humoral counterpart. In the present study, we have analyzed the humoral response of goat kids subjected to primary and challenge infections with Eimeria ninakholyakimovae. Specific levels of IgG and IgM in serum samples and IgA in the ileal mucus were estimated. In infected kids, significantly increased levels of IgG were observed from 3 weeks post infection onwards in addition to an enhancement of specific IgM and secretory IgA levels. A wide range of peptides of sporulated oocyst antigen (SOA) was recognized by specific IgG as determined by immunoblotting. However, no correlations were found between immunoglobulin levels and OPG counts after challenge infection. Overall, these data indicate a significant specific humoral response of E. ninakohlyakimovae-infected goat kids that does not seem to convey immunoprotection. Further studies should be addressed to clarify if the lack of correlation might be associated to the type of antigen used for the immunoenzimatic assays, the age of the animals or other factors.

Protein microarray-mediated detection of antienterovirus antibodies in serum.

ObjectiveTo utilize prokaryotic gene expression and protein microarray to develop and evaluate a sensitive, accurate protein microarray assay for detecting antienterovirus antibodies in serum samples from patients with hand, foot and mouth disease (HFMD). Enterovirus 71 (EV71) and coxsackievirus A16 (CA16), two common causative agents for HFMD, were used for assay development.MethodsSerum was collected from patients with HFMD and healthy controls. EV71 and CA16 VP1 and VP3 genes were expressed in transfected Escherichia coli; the resultant VP1 and 3 proteins were used in a microarray assay for human serum EV71 and CA16 immunoglobulin (Ig) M and IgG. To validate the microarray assay, serum samples were tested for EV71 IgM using enzyme-linked immunosorbent assay (ELISA).ResultsOut of 50 patients with HFMD, EV71 IgM and CA16 IgM was detected in 80% and 44% of serum samples, respectively, using protein microarray, and EV71 IgM was detected in 78% of samples using ELISA. Protein microarray and ELISA showed 100% specificity for EV71-IgM detection.ConclusionThe protein microarray assay developed in the present study shows potential as a sensitive technique for detecting EV71 IgM in serum samples from patients with HFMD.

A study on the status of inflammatory systems in camels naturally infected with Toxoplasma gondii.

Toxoplasma gondii is a unique intracellular parasite with a worldwide distribution. This parasite infects a variety of cells in a wide range of animal species such as dromedary camels (Camelus dromedarius). In order to evaluate the pattern of possible changes in the blood level of some inflammatory mediators and antioxidant enzymes in camels infected with T. gondii, blood samples were taken from a total of 493 dromedary camels and serum concentrations of inflammatory mediators, acute phase proteins and antioxidant enzymes were measured. According to serological data, no seropositivity was found for anti-T. gondii IgM in serum samples; however, 49 camels (9.93%) showed positive titrations for anti-Toxoplasma IgG. The analyses of data in seropositive animals showed significant increases (P < 0.05) in the serum level of IL-1β and adenosine deaminase activity; however, IFN-γ and TNF-α demonstrated no significant changes in serum samples of the infected camels. In addition, while major acute phase proteins (haptoglobin (Hp) and serum amyloid A (SAA)) were markedly elevated in infected camels, the activity of antioxidant enzymes (SOD and GPX) was remarkably decreased in the blood samples of infected animals. Thus, during the chronic infection in camels, T. gondii can promote significant rises in concentrations of some cytokines (such as IL-1β), acute phase proteins and adenosine deaminase.

Seroprevalence of toxoplasmosis and rubella in pregnant women attending antenatal private clinic at Ouagadougou, Burkina Faso

ObjectiveTo evaluate the prevalence of toxoplasmosis and rubella among pregnant women at Ouagadougou in Burkina Faso. MethodsAll patient sera were tested for rubella and toxoplasmosis anti-IgG using commercial ELISA kits (Platelia™ Rubella IgG and Platelia™ Toxo IgG). The presence of anti-rubella and anti-toxoplasmosis IgM in serum samples was tested using commercial ELISA kits Platelia Rubella IgM and Platelia Toxo IgM. ResultsAmong all the pregnant women tested for toxoplasmosis and rubella, their prevalence were 20.3% and 77.0%, respectively. Pregnant women in the age group of 18-25 years showed the highest frequency of anti-toxoplasmosis (34.5%) and anti-rubella IgG (84.6%). The prevalence of anti-toxoplasma and anti-rubella IgG decreased between 2006 and 2008 from 32.7% to 12.1% and 84.6% to 65.0%, respectively. There was no significant association between age and the mean titer of anti-toxoplasmosis IgG among pregnant women. ConclusionsThe diagnosis of toxoplasmosis and rubella is necessary in pregnant women in Burkina Faso because of the low immunization coverage rate of rubella and the high level of exposure to these two infections which can be harmful to the newborn if contracted by women before the third trimester of pregnancy.

Establishment and characterization of a novel genetic hybrid cell line for propagation of four pathogens

Chromosomal DNAs were purified from human epidermoid carcinoma (HEP-2) cells and transfected into human embryonic lung (HEL) cells to establish a genetic hybrid cell line susceptible to infections by toxoplasma, rubella virus, cytomegalovirus, and herpes simplex virus. Karyotype analysis showed that the resultant hybrid cells, designated D3, had a chromosome number of 96, which was stable after passage for 100 generations. Direct microscopy and immunofluorescence showed that the D3 cells could be infected by the four pathogens with overt cytopathic effects. The toxoplasma and three viruses were purified from infected D3 cells by sucrose gradient centrifugation and used as the antigens for detection of specific IgG and/or IgM in serum samples from pregnant women with suspicious infections by the four pathogens, the results of which were consistent with those of commercial kits. These data indicate that a stable genetic hybrid cell line has been generated, which is a valuable tool for the isolation of the four intrauterine pathogens and for the preparation of antigens for serological tests.

Immunoglobulin A Antibody Responses in Dengue Patients: a Useful Marker for Serodiagnosis of Dengue Virus Infection

We determined the usefulness of an immunoglobulin A (IgA) antibody-capture enzyme-linked immunosorbent assay for serodiagnosis of dengue virus infections. The results indicate that the presence of IgA and IgM in serum samples assures recent primary dengue virus infection even with a single serum sample.

Role of transforming growth factor-β in breast milk for initiation of IgA production in newborn infants

Background: Transforming growth factor (TGF)-β has a crucial effect on IgA production, which is the major humoral effector of mucosal immunity. Breast milk contains the abundant amount of TGF-β in the early period of lactation. Aim–study design: To verify the notion that TGF-β in breast milk might contribute to the development of IgA production in newborns, we investigated the association of TGF-β in maternal colostrum with an increase of serum IgA in newborns during the first month of life. Subjects and methods: The concentrations of TGF-β1 and TGF-β2, including IL-6 and IL-10, in colostrum samples from 55 healthy mothers were determined by ELISA. The levels of IgA and IgM in serum samples collected from corresponding newborn babies at birth and at 1 month of age were measured by ELISA. Results: TGF-β1 and TGF-β2 were detected in substantial quantities in all colostrum samples, but IL-6 and IL-10 were present only in a proportion of samples. An increase of serum IgA in newborn during the first month of life was significantly higher than that of serum IgM ( p<0.001). Notably, an increase of serum IgA in newborns during 1 month of life was well correlated with levels of both TGF-β1 ( r=0.38, p=0.005) and TGF-β2 ( r=0.45, p=0.0005) in colostrum, while that of IgM was marginally correlated with colostral TGF-β2 ( r=0.28, p=0.04). The association of increase of serum IgA in newborns with IL-6 and IL-10 in colostrum was not evident. Conclusion: Our findings suggest that TGF-β in colostrum might serve as the starter of IgA production in newborn infants.

Use of RT-PCR on oral fluid samples to assist the identification of measles cases during an outbreak.

This study investigated the occurrence of mild modified measles cases during an outbreak in Niterói, RJ, Brazil by using RT-PCR on oral fluid samples. From August to December 1997 a total of 76 patients with rash were seen at the study sites. Confirmed diagnosis by serology was achieved in 47 cases: measles (39.5%), rubella (13.2%), HHV-6 (3.9%), human parvovirus B19 (3.9%), dengue fever (3%). For 19 of the 29 patients without a conclusive diagnosis paired serum and saliva samples were available for further tests. In four of them, measles virus RNA was detected by RT-PCR in saliva samples in the absence of specific IgM in serum samples. Vaccination histories obtained from three of the RT-PCR positive cases showed that individuals previously immunized can still be infected and contribute to the circulation of measles virus. This study demonstrated the usefulness of RT-PCR on non-invasive clinical samples for the investigation of measles cases.

The effect of timing of sample collection on the detection of measles-specific IgM in serum and oral fluid samples after primary measles vaccination.

This study compares the timing of the rise and decline of measles-specific IgM in serum samples and in oral fluid samples. Two hundred and eighty 9-month-old infants presenting for routine measles vaccination in Addis Ababa, Ethiopia, were enrolled. Paired serum and oral fluid samples were collected before and 1, 2, 3 or 4 weeks after measles vaccination. Samples were tested by using a modified antibody-capture enzyme immunoassay. For the 321 IgM-negative pre- and post-vaccination serum samples, 317 (99 %) of their corresponding oral fluid samples were IgM-negative. Among the 130 IgM-positive serum samples, 75% of their paired oral fluid samples were IgM-positive, with the percentage rising to 87% after oral fluid samples collected > or =3.5 weeks after vaccination were excluded. Among the post-vaccination serum samples, the percent IgM-positive peaked in week 3 and declined to 79% in week 4. For post-vaccination oral fluid samples, the percent IgM-positive peaked in weeks 2 and 3, and then declined to 43% in week 4. This modified antibody-capture enzyme immunoassay appears to detect vaccine-induced measles-specific IgM in the first 3 weeks after vaccination.

A solid-phase reverse immunosorbent test (SPRIST) for the demonstration of specific-mumps virus IgM-class antibody.

A solid-phase reverse ImmunoSorbent Test (SPRIST) was developed for the detection of mumps-specific IgM in serum samples. This technique involves antibody attachment to a solid-phase using anti-human IgM followed by the addition of hemagglutinin and erythrocytes. In a retrospective study on 40 sera, total agreement was obtained between SPRIST and hemagglutination-inhibition after density gradient centrifugation of sera. Neither rheumatoid factor nor heterophil antibody seemed to affect the reaction. This preliminary study was fully confirmed when 2438 sera were prospectively examined; 84 sera yielding positive results originated from patients with clinical symptoms of mumps. SPRIST might offer an interesting alternative to routine detection of recent mumps infections.

The IgM and IgG response to Bordetella pertussis vaccination and infection.

Ultracentrifugation was used to separate IgG from IgM in serum samples from children after pertussis vaccination or infection and the fractions were examined by indirect haemagglutination (IHA) and complement-fixation (CF) tests. IHA detected pertussis-specific IgM and IgG but CF detected only IgG which appeared to be of a different subclass from the IHA IgG. The IgM titres were higher after infection than after vaccination but the reverse tended to apply to the IgG titres. Little IgM or CF IgG was detected 5 months after a dose of vaccine, but the IHA IgG persisted longer. Vaccinated children who were subsequently infected showed IgM and CF IgG responses similar to those of unvaccinated, infected children but the IHA IgG titres reached much higher levels.