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This chapter provides details of serological tests which can be used to detect viral antibodies or antigens in serum, saliva or urine (e.g. ELISA, EIA, IF, CFT, HAI, neutralisation, Western blot, line immunoassays and avidity tests). It details the utility of each test.
In resource-limited countries, the lack of widespread screening masks the true situation of COVID-19. We conducted this study to assess SARS-CoV-2 spread by detection of specific antibodies and to determine associated factors. A population-based cross-sectional study was conducted. Subjects were tested for the presence of two antibodies (IgM and IgG) specific to SARS-CoV-2. Data collection was done using a smartphone with the KoboCollect application. Prevalence of antibodies was estimated with 95% confidence intervals. Logistic regression was used to determine factors associated with positive serological test. A total of 9,094 persons were tested in 4,340 households. The mean age was 30.18 ± 18.65 years, 46.5% male. The overall seroprevalence (prevalence, 95% CI) of SARS-CoV-2 antibodies was (48.2% [47.2%–49.2%]). Being vaccinated, having been in contact with a COVID-19 patient, being older than 50 years, living in a union, having secondary education and having tertiary education were factors independently associated with the likelihood of having anti-sars-CoV-2. We estimate in February 2022 that 48% persons had antibodies against the COVID-19 virus, more among those vaccinated. Vaccination intensification in low prevalence departments will reduce the risk of new outbreaks.
This study aimed to analyse the seroprevalence of SARS-CoV-2 in Kazakhstan. This is a cross-sectional study of adult population in Kazakhstan for the period from October 2021 to May 2022. For the study, 6 720 people aged 18 to 69 were recruited (from 17 regions). The demographic data were collected and analysed. Gender was evenly distributed (males 49.9%, females 50.1%). Women exhibited a higher seroprevalence than men (IgM 20.7% vs 17.9% and IgG 46.1% vs 41.5%). The highest prevalence of IgM was found in the age group of 30–39. However, the highest prevalence of IgG was detected in the age group of 60–69. The seroprevalence of IgG increased across all groups (from 39.7% in 18–29 age groups to 53.1% in 60–69 age groups). The odds for a positive test were significantly increased in older age groups 50–59 (p < 0.0001) and 60–69 (p < 0.0001). The odds of a positive test were 1.12 times higher in females compared to males (p = 0.0294). The odds for a positive test were significantly higher in eight regions (Astana, Akmola, Atyrau, Western Kazakhstan region, Kostanai, Turkestan, Eastern Kazakhstan region, and Shymkent) compared to Almaty city. The odds of a positive test were three times higher in Astana and the Western Kazakhstan region than in Almaty city. In urban areas, the odds of a positive test were 0.75 times lower than in rural areas (p < 0.0001). The study’s results showed an adequate level of seroprevalence (63%) that exceeds the essential minimum of herd immunity indicators in the country. There was significant geographic variability with a higher prevalence of IgG/IgM antibodies to SARS-CoV-2 in rural areas.
1. The innate immune system is immediate and non-specific, whilst the adaptive immune system mounts a slower, specific response to pathogens.
2. The complement system can be activated by either the innate or the adaptive immune system.
3. There are five classes of antibody, with variable roles.
4. Primary immunodeficiency disorders are genetic or congenital in origin, are present early in life and are rare.
5. Secondary acquired immunodeficiency is more common and linked to drug treatment, haematological disease, human immunodeficiency virus infection and critical illness.
Hemolytic disease of the fetus and newborn is a common cause of intrauterine death and increased neonatal morbidity. While this condition may occur as a result of maternal antibody formation to a variety of different antibodies,(alloimmunization), that which occurs in response to the Rhesus D antigen is the most common. Many centers now perform intrauterine fetal blood transfusion, a required mode of therapy for alloimunization, which is necessary to prevent fetal demise. The ability to accurately detect and treat fetuses affected by this condition depends on adequate prenatal care, astute maternal-fetal practitioners, and a robust center with the ability to analyze fetal blood samples which directs transfusion therapy. While intrauterine therapy is not without complications, and is commonly characterized by repeated fetal transfusions, the ability to transfuse fetuses in-utero has drastically improved the prognosis for affected babies.
We report the development of a regression model to predict the prevalence of severe acute respiratory syndrome coronavirus (SARS-CoV-2) antibodies on a population level based on self-reported symptoms. We assessed participant-reported symptoms in the past 12 weeks, as well as the presence of SARS-CoV-2 antibodies during a study conducted in April 2020 in Ischgl, Austria. We conducted multivariate binary logistic regression to predict seroprevalence in the sample. Participants (n = 451) were on average 47.4 years old (s.d. 16.8) and 52.5% female. SARS-CoV-2 antibodies were found in n = 197 (43.7%) participants. In the multivariate analysis, three significant predictors were included and the odds ratios (OR) for the most predictive categories were cough (OR 3.34, CI 1.70–6.58), gustatory/olfactory alterations (OR 13.78, CI 5.90–32.17) and limb pain (OR 2.55, CI 1.20–6.50). The area under the receiver operating characteristic curve was 0.773 (95% CI 0.727–0.820). Our regression model may be used to estimate the seroprevalence on a population level and a web application is being developed to facilitate the use of the model.
Tick-borne encephalitis (TBE) is a vector-borne infection associated with a variety of potentially serious complications and sequelae. Vaccination against TBE is strongly recommended for people living in endemic areas. There are two TBE vaccination schemes – standard and rapid – which differ in the onset of protection. With vaccination in a rapid schedule, protection starts as early as 4 weeks after the first dose and is therefore especially recommended for non-immune individuals travelling to endemic areas. Both schemes work reliably in immunocompetent individuals, but only little is known about how TBE vaccination works in people with HIV infection. Our aim was to assess the immunogenicity and safety of the rapid scheme of TBE vaccination in HIV-1 infected individuals. Concentrations of TBE-specific IgG > 126 VIEU/ml were considered protective. The seroprotection rate was 35.7% on day 28 and 39.3% on day 60. There were no differences between responders and non-responders in baseline and nadir CD4 + T lymphocytes. No serious adverse events were observed after vaccination. The immunogenicity of the TBE vaccination was unsatisfactory in our study and early protection was only achieved in a small proportion of vaccinees. Therefore, TBE vaccination with the rapid scheme cannot be recommended for HIV-1 infected individuals.
Bone marrow (BM) trephine biopsy (BMB) is a frequent and routine diagnostic investigation, as nicely described in the previous chapters, and is also widely used for follow-up of haematological disorders to judge the effectiveness of therapeutic interventions [1]. In addition, several drugs applied primarily for the treatment of non-haematological disorders may cause serious haematological side effects, such as pancytopaenia, agranulocytosis or anaemia. On occasion, the causative link between the haematological symptoms and the previous drug exposure is not perceived and it is not uncommon for BMB to be obtained in such circumstances without any information of a history of previous drug exposure being made available to the haematopathologist [2].
Determination of antibodies against ToRCH antigens at the beginning of pregnancy allows assessment of both the maternal immune status and the risks to an adverse pregnancy outcome. Age-standardised seroprevalences were determined in sera from 1009 women of childbearing age residing in Mexico, Brazil, Germany, Poland, Turkey or China using a multiparametric immunoblot containing antigen substrates for antibodies against Toxoplasma gondii, rubella virus, cytomegalovirus (CMV), herpes simplex viruses (HSV-1, HSV-2), Bordetella pertussis, Chlamydia trachomatis, parvovirus B19, Treponema pallidum and varicella zoster virus (VZV). Seroprevalences for antibodies against HSV-1 were >90% in samples from Brazil and Turkey, whereas the other four countries showed lower mean age-adjusted seroprevalences (range: 62.5–87.9%). Samples from Brazilian women showed elevated seroprevalences of antibodies against HSV-2 (40.1%), C. trachomatis (46.8%) and B. pertussis (56.6%) compared to the other five countries. Seroprevalences of anti-T. gondii antibodies (0.5%) and anti-parvovirus B19 antibodies (7.5%) were low in samples from Chinese women, compared to the other five countries. Samples from German women revealed a low age-standardised seroprevalence of anti-CMV antibodies (28.8%) compared to the other five countries. These global differences in immune status of women in childbearing age advocate country-specific prophylaxis strategies to avoid infection with ToRCH pathogens.
Borrelia burgdorferi (Bb) infection can affect the central nervous system and possibly lead to psychiatric disorders. We compared clinical and demographic variables in Bb seropositive and seronegative psychiatric patients and healthy controls.
Method.
Nine hundred and twenty-six consecutive psychiatric patients were screened for antibodies to Bb and compared with 884 simultaneously recruited healthy subjects.
Results.
Contrary to healthy controls, seropositive psychiatric patients were significantly younger than seronegative ones. None of the studied psychiatric diagnostic categories exhibited stronger association with seropositivity. There were no differences between seropositive and seronegative psychiatric patients in hospitalization length, proportion of previously hospitalized patients and proportion of subjects with family history of psychiatric disorders.
Conclusion.
These findings elaborate on potential association between Bb infection and psychiatric morbidity, but fail to identify any specific clinical ‘signature’ of Bb infection.
The past decade has seen a surge of reports and investigations into cases of autoimmune-mediated encephalitis. The increasing recognition of these disorders is especially of relevance to the fields of neurology and psychiatry. Autoimmune encephalitis involves antibodies against synaptic receptors, neuronal cell surface proteins and intracellular targets. These disorders feature prominent symptoms of cognitive impairment and behavioural changes, often associated with the presence of seizures. Early in the clinical course, autoimmune encephalitis may manifest as psychiatric symptoms of psychosis and involve psychiatry as an initial point of contact. Although commonly associated with malignancy, these disorders can present in the absence of an inciting neoplasm. The identification of autoimmune encephalitis is of clinical importance as a large proportion of individuals experience a response to immunotherapy. This review focuses on the current state of knowledge on n-methyl-d-aspartate (NMDA) receptor-associated encephalitis and limbic encephalitis, the latter predominantly involving antibodies against the α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) receptor, the γ-aminobutyric acid (GABA)B receptor and leucine-rich glioma-inactivated 1 (LGI1) protein. In addition, we briefly describe anti-dopamine D2 receptor encephalitis. A summary of the literature will focus on common clinical presentations and course, diagnostic approaches and response to treatment. Since a substantial proportion of patients with autoimmune encephalitis exhibit symptoms of psychosis, the relevance of this disorder to theories of psychosis and schizophrenia will also be discussed.
Why do hosts vary so much in parasite burden, how does this variation translate to variation in host demographic rates and parasite transmission, and how does varied transmission intensity impact selection upon immune defence of individuals? The theoretical foundations of disease ecology provide predictions for the answers to these questions, yet testing such predictions with empirical data poses many challenges. We show how the long-term ecological and genetic study of the unmanaged Soay sheep of St Kilda has addressed fundamental questions in disease ecology, with longitudinal data on parasite burden, immune defence, condition, survival, and fecundity of >10,000 individuals. The rich individual-scale data are complemented by >30 years of data on sheep population dynamics and genetic diversity as well as parasite dynamics and diversity. Population-scale work has documented the range of parasite species present and the contribution of the most prevalent and virulent parasites to regulating sheep dynamics. Individual-scale work has identified drivers of variation in parasite burden and tested hypotheses about costs and benefits of defence in a quest to determine how natural selection has shaped immune function of the sheep.
The relationship between epilepsy and the presence of visceral larva migrans caused by Toxocara canis in Mexican children remains uncertain; however, this relationship needs to be elucidated because these parasite larvae can invade the human central nervous system. Accordingly, this study aimed to determine the frequency and specificity of anti-T. canis antibodies in the sera of children with epilepsy to determine the relationship between this parasite and epilepsy. The sera samples of 214 children were examined: 111 children diagnosed with epilepsy and 103 clinically healthy children without neurological disorders. In the sera of each group, the presence and specificity of anti-T. canis and anti-Ascaris lumbricoides antibodies, as well as the cross-reactivity between them, were assessed using enzyme-linked immunosorbent assay and Western blotting analysis. Among the children with epilepsy, 25.2% exhibited seropositivity to T. canis. Cross-reactivity against the A. lumbricoides antigen was present in 46.8% of the children with epilepsy, whereas 11.7% of the children with epilepsy and anti-T. canis antibodies did not exhibit cross-reactivity against this antigen. The Western blotting analysis of the sera from the children with epilepsy demonstrated the presence of T. canis proteins, with molecular weights of 24, 35, 55, 70, 120 and 210 kDa, and A lumbricoides proteins with molecular weights of 70, 80 and 110 kDa. Our results revealed the presence of anti-T. canis antibodies in the children with epilepsy; furthermore, cross-reactivity tests with A. lumbricoides showed the importance of the presence of anti-T. canis antibodies in revealing the relationship between this parasite and epilepsy in children.
Outbreaks of Old World cutaneous leishmaniasis (CL) have significantly increased due to the conflicts in the Middle East, with most of the cases occurring in resource-limited areas such as refugee settlements. The standard methods of diagnosis include microscopy and parasite culture, which have several limitations. To address the growing need for a CL diagnostic that can be field applicable, we have identified five candidate neoglycoproteins (NGPs): Galα (NGP3B), Galα(1,3)Galα (NGP17B), Galα(1,3)Galβ (NGP9B), Galα(1,6)[Galα(1,2)]Galβ (NGP11B), and Galα(1,3)Galβ(1,4)Glcβ (NGP1B) that are differentially recognized in sera from individuals with Leishmania major infection as compared with sera from heterologous controls. These candidates contain terminal, non-reducing α-galactopyranosyl (α-Gal) residues, which are known potent immunogens to humans. Logistic regression models found that NGP3B retained the best diagnostic potential (area under the curve from receiver-operating characteristic curve = 0.8). Our data add to the growing body of work demonstrating the exploitability of the human anti-α-Gal response in CL diagnosis.
Malaria is one the most serious infectious diseases with over 200 million clinical cases annually. Most cases of the severe disease are caused by Plasmodium falciparum. The blood stage of Plasmodium parasite is entirely responsible for malaria-associated pathology. The population most susceptible to severe malaria are children under the age of 5, with low levels of immunity. It is only after many years of repeated exposure that individuals living in endemic areas develop clinical immunity. This form of protection prevents clinical episodes by substantially reducing parasite burden. Naturally acquired immunity predominantly targets blood-stage parasites with antibody responses being the main mediators of protection. The targets of clinical immunity are the extracellular merozoite and the infected erythrocyte surface, with the extremely diverse PfEMP1 proteins the main target here. This observation provides a strong rationale that an effective anti-malaria vaccine targeting blood-stage parasites is achievable. Thus the identification of antigenic targets of naturally acquired immunity remains an important step towards the formulation of novel vaccine combinations before testing their efficacy in clinical trials. This review summarizes the main findings to date defining antigenic targets present on the extracellular merozoite associated with naturally acquired immunity to P. falciparum malaria.
There has been much recent excitement about the possibility that some cases of psychosis may be wholly due to brain-reactive antibodies, with antibodies to N-methyl-D-aspartate receptor (NMDAR) and the voltage-gated potassium channel (VGKC)-complex reported in a few patients with first-episode psychosis (FEP).
Methods
Participants were recruited from psychiatric services in South London, UK, from 2009 to 2011 as part of the Genetics and Psychosis study. We conducted a case–control study to examine NMDAR and VGKC-complex antibody levels and rates of antibody positivity in 96 patients presenting with FEP and 98 controls matched for age and sex. Leucine-rich glioma inactiviated-1 (LGI1) and contactin-associated protein (CASPR) antibodies were also measured. Notably, patients with suspicion of organic disease were excluded.
Results
VGKC-complex antibodies were found in both cases (n = 3) and controls (n = 2). NMDAR antibody positivity was seen in one case and one control. Either LGI1-Abs or CASPR2-Abs were found in three cases and three controls. Neuronal antibody staining, consistent with the above results or indicating potential novel antigens, was overall positive in four patients but also in six controls. Overall, antibody positivity was at low levels only and not higher in cases than in controls.
Conclusions
This case–control study of the prevalence of antibodies in FEP does not provide evidence to support the hypothesis that FEP is associated with an immune-mediated process in a subgroup of patients. Nevertheless, as other bio-clinical factors may influence the effect of such antibodies in a given individual, and patients with organic neurological disease may be misdiagnosed as FEP, the field requires more research to put these findings in context.
Immunochemical techniques offer many advantages over chromatographic methods used for pesticide trace analysis of substrates such as soil, water, plants, urine, and blood. These advantages include speed of processing samples, high specificity for detecting a pesticide, reduced amount of preparation and cleanup of the sample before analysis, and a dramatic increase in the number of samples that can be analyzed. Immunoassays are based on the principle that antibodies to pesticides can be prepared, in animals, that can recognize and attach with exquisite specificity to certain chemical configurations displayed on the surface of a molecule. Small molecules such as herbicides usually are not immunogenic but can be made so by chemically bonding them to a large immunogenic protein such as bovine serum albumin before injection into an animal. Development of herbicide-specific antibodies and their use in direct and indirect enzyme-linked immunosorbent assays (EIA or ELISA) as well as radioimmunoassays (RIA) are discussed. The principles behind monoclonal antibody production are outlined, and immunoassays using polyclonal and monoclonal antibodies are compared. Specific reference is made to the development and use of indirect ELISA and RIA procedures for trace analysis of 2,4-D and picloram.
We explored age-dependent patterns in haemagglutination inhibition (HI) titre to seasonal [1956 A(H1N1), 1977 A(H1N1), 2007 A(H1N1)] and pandemic [A(H1N1)pdm09] influenza strains using serological data collected from an adult French influenza cohort. Subjects were recruited by their general practitioners from 2008 to 2009 and followed until 2010. We explored age-related differences between strain-specific HI titres using 1053 serological samples collected over the study period from 398 unvaccinated subjects. HI titres against the tested seasonal and pandemic strains were determined using the HI technique. Geometric mean titres (GMTs) were estimated using regression models for interval-censored data. Generalized additive mixed models were fit to log-transformed HI estimates to study the relationship between HI titre and age (age at inclusion and/or age at initial strain circulation). GMT against one strain was consistently highest in the birth cohort exposed to that strain during childhood, with peak titres observed in subjects aged 7–8 years at the time of initial strain circulation. Our results complete previous findings on influenza A(H3N2) strains and identify a strain-dependent relationship between HI titre and age at initial strain circulation.
Autoimmune encephalitis associated with anti-neuronal antibodies may be challenging to distinguish from primary psychiatric disorders. The significance of anti-neuronal antibodies in psychiatric patients without clear evidence of autoimmune encephalitis is unknown. We investigated the serum prevalence of six anti-neuronal autoantibodies in a cohort of unselected patients admitted to acute psychiatric care.
Method
Serum was drawn from 925 patients admitted to acute psychiatric in-patient care. Psychiatric diagnoses were set according to International Classification of Diseases (ICD)-10 criteria. Antibody analysis was performed with an indirect immunofluorescence test for N-methyl d-aspartate receptor (NMDAR) antibodies and five other anti-neuronal autoantibodies of the immunoglobulin (Ig) classes IgA, IgG and IgM isotype.
Results
Anti-neuronal autoantibodies were found in 11.6% of patients: NMDAR antibodies in 7.6%, contactin-associated protein-like 2 (CASPR2) antibodies in 2.5%, glutamic acid decarboxylase-65 (GAD65) antibodies in 1.9%, and α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) receptor antibodies in 0.1%. Leucine-rich glioma-inactivated protein-1 (LGI1) and γ-aminobutyric acid B (GABAB) receptor antibodies were not detected. NMDAR antibodies of class IgG were present in five patients only (0.5%). NMDAR antibodies of all Ig classes were equally prevalent in patients with and without psychosis. There were no significant differences in antibody prevalence in the different diagnostic categories, except for a higher odds ratio of being NMDAR antibody positive for patients without a specific psychiatric diagnosis.
Conclusions
NMDAR IgG autoantibodies, which are known to be strongly associated with anti-NMDAR encephalitis, were rarely found. CASPR2 and GAD65 antibodies were more frequently encountered in the present study than previously reported. Further research on the clinical significance of anti-neuronal autoantibodies in patients with acute psychiatric symptoms is needed.