Genetic mutations and immunological features of severe combined immunodeficiency patients in Iran.

BACKGROUND: Severe combined immunodeficiency (SCID) is the most severe form of primary immunodeficiency disorders that is characterized by impaired early T lymphocyte differentiation and is variably associated with abnormal development of other lymphocyte lineages. SCID can be caused by mutations in more than 20 different genes. Molecular diagnosis in SCID patients contributes to genetic counseling, prenatal diagnosis, treatment modalities, and overall prognosis. In this cohort, the clinical, laboratory and genetic data related to Iranian SCID patients were comprehensively evaluated and efficiency of stepwise sequencing methods approach based on immunophenotype grouping was investigated METHODS: Clinical and laboratory data from 242 patients with SCID phenotype were evaluated. Molecular genetic analysis methods including Sanger sequencing, targeted gene panel and whole exome sequencing were performed on 62 patients. RESULTS: Mortality rate was 78.9% in the cohort with a median follow-up of four months. The majority of the patients had a phenotype of T-NK-B+ (34.3%) and the most severe clinical manifestation and highest mortality rate were observed in T-NK-B- SCID cases. Genetic mutations were confirmed in 50 patients (80.6%), of which defects in recombination-activating genes (RAG1 and RAG2) were found in 16 patients (32.0%). The lowest level of CD4+ and CD8+ cells were observed in patients with ADA deficiency (p = 0.026) and IL2RG deficiency (p = 0.019), respectively. CONCLUSION: Current findings suggest that candidate gene approach based on patient's immunophenotype might accelerate molecular diagnosis of SCID patients. Candidate gene selection should be done according to the frequency of disease-causing genes in different populations. Targeted gene panel, WES and WGS methods can be used for the cases which are not diagnosed using this method.

Clinical Manifestations, Immunological Characteristics and Genetic Analysis of Patients with Hyper-Immunoglobulin M Syndrome in Iran.

BACKGROUND: Hyper-immunoglobulin M (HIGM) syndrome is a rare heterogeneous group of primary immunodeficiency disorders characterized by low or absent serum levels of IgG and IgA along with normal or elevated serum levels of IgM. METHODS: Clinical and immunological data were collected from the 75 patients' medical records diagnosed in Children's Medical Center affiliated to Tehran University Medical Sciences and other Universities of Medical Sciences in Iran. Among 75 selected patients, 48 patients (64%) were analyzed genetically using targeted and whole-exome sequencing. RESULTS: The ratio of male to female was 2.9:1. The median age at the onset of the disease, time of diagnosis, and diagnostic delay were 10.5, 50, and 24 months, respectively. Pneumonia and lower respiratory tract infections (61.3%) were the most common complications. Responsible genes were identified in 35 patients (72.9%) out 48 genetically analyzed patients. Cluster of differentiation 40 ligand gene was the most mutated gene observed in 24 patients (68.5%) followed by activation-induced cytidine deaminase gene in 7 patients, lipopolysaccharide-responsive and beige-like anchor (1 patient), nuclear factor-kappa-B essential modulator (1 patient), phosphoinositide-3-kinase regulatory subunit 1 (1 patient), and nuclear factor kappa B subunit 1 (1 patient) genes. Nineteen (25.3%) patients died during the study period, and pneumonia was the major cause of death occurred in 6 (31.6%) patients. CONCLUSION: Physicians in our country should carefully pay attention to respiratory tract infections and pneumonia, particularly in patients with a positive family history. Further investigations are required for detection of new genes and pathways resulting in HIGM phenotype.

Comparison of Common Monogenic Defects in a Large Predominantly Antibody Deficiency Cohort.

BACKGROUND: Predominantly antibody deficiencies (PADs) are the most common primary immunodeficiencies, characterized by hypogammaglobulinemia and inability to generate effective antibody responses. OBJECTIVE: We intended to report most common monogenic PADs and to investigate how patients with PAD who were primarily diagnosed as suffering from agammaglobulinemia, hyper-IgM (HIgM) syndrome, and common variable immunodeficiency (CVID) have different clinical and immunological findings. METHODS: Stepwise next-generation sequencing and Sanger sequencing were performed for confirmation of the mutations in the patients clinically diagnosed as suffering from agammaglobulinemia, HIgM syndrome, and CVID. RESULTS: Among 550 registered patients, the predominant genetic defects associated with agammaglobulinemia (48 Bruton's tyrosine kinase [BTK] and 6 µ heavy chain deficiencies), HIgM syndrome (21 CD40 ligand and 7 activation-induced cytidine deaminase deficiencies), and CVID (17 lipopolysaccharides-responsive beige-like anchor deficiency and 12 atypical Immunodeficiency, Centromeric instability, and Facial dysmorphism syndromes) were identified. Clinical disease severity was significantly higher in patients with µ heavy chain and CD40 ligand mutations compared with patients with BTK (P = .003) and activation-induced cytidine deaminase (P = .009) mutations. Paralysis following live polio vaccination was considerably higher in patients with µ heavy chain deficiency compared with BTK deficiency (P < .001). We found a genotype-phenotype correlation among patients with BTK mutations regarding clinical manifestation of meningitis and chronic diarrhea. Surprisingly, we noticed that first presentations in most patients with Immunodeficiency, Centromeric instability, and Facial dysmorphism were respiratory complications (P = .008), whereas first presentations in patients with lipopolysaccharides-responsive beige-like anchor deficiency were nonrespiratory complications (P = .008). CONCLUSIONS: This study highlights similarities and differences in the clinical and genetic spectrum of the most common PAD-associated gene defects. This comprehensive comparison will facilitate clinical decision making, and improve prognosis and targeted treatment.

Fourth Update on the Iranian National Registry of Primary Immunodeficiencies: Integration of Molecular Diagnosis.

BACKGROUND: The number of inherited diseases and the spectrum of clinical manifestations of primary immunodeficiency disorders (PIDs) are ever-expanding. Molecular diagnosis using genomic approaches should be performed for all PID patients since it provides a resource to improve the management and to estimate the prognosis of patients with these rare immune disorders. METHOD: The current update of Iranian PID registry (IPIDR) contains the clinical phenotype of newly registered patients during last 5 years (2013-2018) and the result of molecular diagnosis in patients enrolled for targeted and next-generation sequencing. RESULTS: Considering the newly diagnosed patients (n = 1395), the total number of registered PID patients reached 3056 (1852 male and 1204 female) from 31 medical centers. The predominantly antibody deficiency was the most common subcategory of PID (29.5%). The putative causative genetic defect was identified in 1014 patients (33.1%) and an autosomal recessive pattern was found in 79.3% of these patients. Among the genetically different categories of PID patients, the diagnostic rate was highest in defects in immune dysregulation and lowest in predominantly antibody deficiencies and mutations in the MEFV gene were the most frequent genetic disorder in our cohort. CONCLUSIONS: During a 20-year registration of Iranian PID patients, significant changes have been observed by increasing the awareness of the medical community, national PID network establishment, improving therapeutic facilities, and recently by inclusion of the molecular diagnosis. The current collective study of PID phenotypes and genotypes provides a major source for ethnic surveillance, newborn screening, and genetic consultation for prenatal and preimplantation genetic diagnosis.

Quality of Life in Children with Asthma versus Healthy Children.

BACKGROUND: Asthma is the most prevalent chronic disease in the pediatric age group. The disease affects different aspects of the children's lives, such as physical, emotional, social and educational aspects. Thus, more focus has been on the quality of life in these patients rather than the duration of their illness in recent years. AIM: This study examined the different aspects of quality of life in asthmatic children for the first time in this geographic area. METHODS: The study was cross-sectional conducted in 2015-2016. The asthmatic group was 100 patients aged 8 to 12 admitted to the Asthma and Allergy Clinic of Ghaem Hospital (as) in Mashhad with the control group composed of 100 healthy children of the same age and gender. The standard questionnaire pedsQLTM was used for comparing the quality of life of children in the two groups. Statistical analysis was SPSS23 with P-value less than 0.05, which was statistically significant. RESULTS: In each group, 58 patients were boys, and 42 were girls. In a comparison of the quality of life of children, the asthma group with a mean total score of Peds QL 20.99 ± 12.54 compared to the healthy children with a mean total score of Peds QL of 8.8 ± 5.41 had a lower quality of life (P < 0.001). Moreover, regarding various aspects of quality of life asthma group had a lower quality of life in physical performance, emotional performance and performance in school (P < 0.001). Nonetheless, there was no significant difference between the two groups considering social function (P = 0.267). Examining the relationship between Peds QL score of patients with asthma with various variables was indicative of the fact that Peds QL scores were significantly correlated with the gender of the patients, showing better quality of life in the girls (P = 0.001). CONCLUSION: The results indicated that children with asthma have a significantly lower quality of life compared with healthy children of the same age. Also, in examining the different aspects of quality of life, these children had a lower quality of life in physical performance, emotional performance, and performance at school, and were at the level as that of healthy children only in social performance.

A survey of anaphylaxis etiology and treatment.

Identifying the causes of anaphylaxis which is an acute, potentially fatal systemic reaction is very important in every community. Treatment strategies and pitfalls should also be determined. We sought to determine the most common triggers of anaphylaxis, clinical manifestations and treatment strategies in Mashhad, northeast of Iran. An observational cross-sectional study was conducted to evaluate all patients with a history of anaphylactic reaction who were referred to University Allergy Clinics between 2006 and 2016 in Mashhad Iran. We used a combination of patient's clinical history and allergy diagnostic testing including radioallergosorbant test and skin prick test in order to determine the etiology of anaphylaxis. We identified 172 anaphylactic reactions in 70 patients. Median age was 15 years with a range from 6 months to 48 years. The triggers included: foods, 61.4%; drugs, 15.7%; hymenoptera venom, 8.6%; idiopathic, 5.7%; immunotherapy, 4.3% and other etiologies: 5.7%. Nuts and seeds were the most important triggers of food induced anaphylaxis, especially in school children, adolescents and young adults, followed by fruits. However, Cow's milk and hen's egg were the main triggers of anaphylaxis in children aged under 2 years. The most common symptoms were cutaneous and cardiovascular. Corticosteroids (94.3%) and/or antihistamines (85.7%) were used most frequently for treatment followed by intravenous fluids (54.3%), whereas epinephrine was only used in 17.1% of the cases. Food related anaphylaxis and other typical triggers of anaphylaxis are age dependent and the risks and triggers change with age. Epinephrine injection should be increased by improving the awareness of physician and medical teams. The study was approved by the Ethics Committee of the Faculty of Medicine of Mashhad University of Medical Sciences (approved number: IR.MUMS.REC.1393.960).

Evaluation of anxiety and depression in mothers of children with asthma.

BACKGROUND: Asthma is the most common chronic disease in childhood. Parents have an important role in managing asthma in children. Studies have shown a higher degree of depression and anxiety and lower family performance in mothers of asthmatic children in comparison with the control group. OBJECTIVE: The aim of this study was to evaluate the parenting styles and also depression, anxiety and stress parameters in mothers of children with asthma. METHODS: This case-control study was performed on 45 mothers of 3 to 15 years old asthmatic children in the allergy clinic of Mashhad University of Medical Sciences, Mashhad, Iran, during the years of 2014 to 2016. The control group was 45 mothers of non-asthmatic children who were matched for the age of their children with the case group in the same population. The parenting styles, as well as depression and anxiety of mothers were evaluated using parenting scales, and the depression-anxiety-stress scales (DASS). The mothers were also asked to fill a strengths and difficulties questionnaire (SDQ) for their children. Furthermore, parenting styles in the case group were compared to mothers of children without asthma as the control group. The data were then analyzed by SPSS 11.5, using Chi-square, ANOVA, and independent-samples t-test. RESULTS: The results of this study showed that 21 mothers (74.6%) were normal, but 12 mothers (26.7%) had a mild -, 9 (20%) a moderate - and 3 (6.7%) a severe degree of abnormality according to DASS. Independent-samples t-test showed a significant difference between the case and control groups regarding depression in mothers and laxness (p<0.001), over reactivity (p<0.013) and verbosity (p<0.031) in children with asthma. CONCLUSION: The results of this study demonstrated that anxiety and depression are partially frequent in mothers of children with asthma, and parenting styles are less affective in these families.

Epidemiological survey of pediatric food allergy in Mashhad in Northeast Iran.

INTRODUCTION: Food allergy is an increasing problem worldwide, but the foods responsible for food allergy are not the same in different countries, probably because of the role of genetic, cultural, and nutritional factors. The aim of this study was to determine the common food allergens in pediatric patients with different presentation of food allergy. METHODS: In this cross-sectional study, all of the patients were referred to pediatric allergy clinics affiliated with Mashhad University of Medical Sciences from September 2012 to August 2014. For patients with IgE-mediated food allergy that was diagnosed with clinical manifestations, the skin prick test was done. The results were analyzed by SPSS version 17 and statistical analysis was done with the chi-squared test and the t-test. P values < 0.05 were considered statistically significant. RESULTS: Three hundred seventy-one patients (53.9% male, 46.1% female) with ages in the range of three months to 18 years were studied. The most frequent food allergen in all patients with decreasing prevalence were egg white (17.8%), pepper (15.8%), curry (14.3%), egg yolk (14%), cow's milk (10%), and tomato (7.8%). The most common presenting symptoms were respiratory (allergic rhinitis 45%, asthma 32%), dermatologic (atopic dermatitis 30%, urticaria 8.3%), colitis (17.5%), and gasteroesophagial reflux disease (GERD) (2%). According to the prevalence of food allergens in different age groups, we realized that, after the age of three years, the frequency of sensitization to egg white, egg yolk, cow's milk, wheat and cereals was decreased and allergy to pepper and curry was increased. CONCLUSION: The prevalence of culprit foods that produce food allergies depends on several factors, including age, presenting manifestation, and where the patient lives. As many food allergies are outgrown, patients should be reevaluated regularly to determine whether they have lost their reactivity or not.

Teaching final-year medical students in a paediatric ambulatory care unit.

BACKGROUND: The importance of community-based ambulatory experiences for medical students has been emphasised in the past few decades. Although such teaching programmes are assumed to be better for medical students, there is little evidence comparing the out-patient and in-patient experiences in student education. OBJECTIVE: We carried out a study to compare the educational experiences between two in-patient and out-patient clinical units. METHODS: Over a 12-week paediatric clerkship, 38 senior medical students attending the paediatric ward were divided into two groups. One group attended a 15-day paediatric out-patient clinic while the other group attended the in-patient programme. RESULTS: Those who took part in the out-patient clinic programme obtained better scores in a test on common paediatric ambulatory problems when compared with the students who exclusively attended the in-patient teaching programme. The former group all agreed that this ambulatory paediatric course was a beneficial learning experience and consistent with their future career needs.

Primary immunodeficiency disorders in Iran: update and new insights from the third report of the national registry.

BACKGROUND: Primary immunodeficiency disorders (PID) are a group of heterogeneous disorders mainly characterized by severe and recurrent infections and increased susceptibility to malignancies, lymphoproliferative and autoimmune conditions. National registries of PID disorders provide epidemiological data and increase the awareness of medical personnel as well as health care providers. METHODS: This study presents the demographic data and clinical manifestations of Iranian PID patients who were diagnosed from March 2006 till the March of 2013 and were registered in Iranian PID Registry (IPIDR) after its second report of 2006. RESULTS: A total number of 731 new PID patients (455 male and 276 female) from 14 medical centers were enrolled in the current study. Predominantly antibody deficiencies were the most common subcategory of PID (32.3 %) and were followed by combined immunodeficiencies (22.3 %), congenital defects of phagocyte number, function, or both (17.4 %), well-defined syndromes with immunodeficiency (17.2 %), autoinflammatory disorders (5.2 %), diseases of immune dysregulation (2.6 %), defects in innate immunity (1.6 %), and complement deficiencies (1.4 %). Severe combined immunodeficiency was the most common disorder (21.1 %). Other prevalent disorders were common variable immunodeficiency (14.9 %), hyper IgE syndrome (7.7 %), and selective IgA deficiency (7.5 %). CONCLUSIONS: Registration of Iranian PID patients increased the awareness of medical community of Iran and developed diagnostic and therapeutic techniques across more parts of the country. Further efforts must be taken by increasing the coverage of IPIDR via electronically registration and gradual referral system in order to provide better estimation of PID in Iran and reduce the number of undiagnosed cases.

Evaluation of interleukin 1ß in febrile convulsion.

Febrile convulsion (FC) is the most common type of seizure in childhood that occurs in 2-5 % of the children younger than 6 years. Interleukin 1ß (IL-1ß) is a cytokine that contributes to febrile inflammatory responses. There are conflicting results on increasing this cytokine in serum during FC. Thus we measured IL-1ß in febrile children with or without seizure. 60 febrile children (6 months to 5 years old) were divided in two groups, one group consisted of 30 children with FC, the other group consisting of 30 children without seizure which served as control. Blood samples were collected from members of both groups and serum samples were prepared. Interleukin 1ß concentrations were measured using a commercial enzyme-linked immunosorbent assay (ELISA) kit. We found that there was a difference in serum levels of interleukin 1ß between FC and control group but it was not significant. This result may be due to the low number of samples or the result of interleukin 1ß binding to some large proteins such as α2-macroglobolin, complement and soluble type 2 Interleukin 1 receptor, that affected the free interleukin 1ß concentration.We could not find a significant relationship between serum interleukin 1ß concentration and FC.

Molecular analysis of four cases of chronic granulomatous disease caused by defects in NCF-2: the gene encoding the p67-phox.

Chronic granulomatous disease (CGD), a rare inherited primary immunodeficiency disorder, is caused by mutation in any one of the genes encoding components of nicotinamide adenine dinucleotide phosphate (NADPH)-oxidase enzyme. NCF2 gene (encoding P67-phox component) is one of them and its mutation is less common to cause CGD (around 5-6%). Here, we assessed mutation analysis of NCF2 in 4 CGD patients with p67-phox defect in Iran. These patients showed classical CGD symptoms. NCF2 sequence analyses revealed two different homozygous mutations including a nonsense mutation in exon 4, c.304C>T (Arg 102X) in one case and a CA deletion in exon 13 (Leu346fsX380) in one brother and sister; the latter is a new mutation which has not been reported in previous studies. In another patient in whom the attempts to amplify exon 2 individually from genomic DNA were unsuccessful, PCR amplification of exon 2 revealed no band of this exon on agarose gel. A PCR amplification mix of exon 2 and exon 7, with an internal control, confirmed the lack of exon 2 in this patient. Although a gross deletion in other exons of NCF2 has been previously reported, a large deletion encompassing exon 2 has been not reported yet. This abstract was also presented in ESID 2012, Florence, Italy.

Inheritance pattern and clinical aspects of 93 Iranian patients with chronic granulomatous disease.

BACKGROUND: Chronic granulomatous disease (CGD) is a rare immunodeficiency due to a genetic defect in one of the NADPH-oxidase components. We studied CGD inheritance forms (autosomal recessive (AR) or X-linked (XL)) and AR-CGD subtypes in Iran. METHODS: Clinical and functional investigations were conducted in 93 Iranian CGD patients from 75 families. RESULTS: Most of the patients were AR-CGD (87.1%). This was related to consanguineous marriages (p = 0.001). The age of onset of symptoms and diagnosis were lower in XL-CGD compared with AR-CGD (p < 0.0001 for both). Among AR-CGD patients, p47phox defect was the predominant subtype (55.5%). The most common clinical features in patients were lymphadenopathy (65.6%) and pulmonary involvement (57%). XL-CGD patients were affected more frequently with severe infectious manifestations. CONCLUSIONS: Although XL-CGD is the most common type of the disease worldwide, only 12 patients (12.9%) were XL-CGD in our study. The relatively high frequency of AR-CGD is probable due to widely common consanguineous marriages in Iran.

Pistachio allergy-prevalence and in vitro cross-reactivity with other nuts.

BACKGROUND: Tree nut allergy is characterized by a high frequency of life-threatening reactions and is typically lifelong persistent. Some people with a pistachio nut allergy, which is common in the pistachio rich area of Iran, develop a hypersensitivity to other tree nuts as well. The aim of this study was to investigate the prevalence of pistachio nut allergy in Iran, the major pistachio cultivation region in the world. The study also addressed the presence of allergenic cross-reactivity between pistachio and other nuts, including almond, peanut, and cashew in pistachio allergic patients. METHODS: A survey was conducted to determine whether the prevalence of pistachio allergy is affected by exposure to this nut in pistachio cultivation regions, as well as possible cross-reactivity between pistachio and other nuts including cashew, almond, and peanut. Inhibition Western blot and inhibition ELISA studies were conducted to assess the presence of allergenic cross-reactivity between pistachio and the other tree nuts. RESULTS: Our results revealed that the prevalence of pistachio allergy is twice as much in pistachio cultivation regions than other areas. Western blotting and inhibition ELISA presented high percentages of inhibition with pistachio and cashew, followed by almond and, to some degree, peanut which indicates different levels of allergenic cross-reactivity. CONCLUSIONS: The results indicate that exposure of people to pistachio significantly affects the prevalence of its allergic reactions. In addition, it was observed that, among pistachio allergic subjects, such exposure may affect the co-sensitivities with other nuts, including cashew and almond. The plant taxonomic classification of pistachio and other tree nuts does appear to predict allergenic cross-reactivity.

Successful allogeneic stem cell transplantation with a reduced-intensity conditioning in a leukocyte adhesion deficiency type I patient.

LAD-I is a rare, autosomal recessive, primary immunodeficiency in which phagocyte adhesion and chemotaxis are impaired. Multiple infections in the absence of pus accumulation and persistent elevated peripheral blood neutrophil counts are the hallmark of LAD-I. Allogeneic HSCT is the only treatment proved to be potentially curative for phagocyte adhesion impairment in LAD-I. Here, we report on a case of a 30-month-old girl with LAD-I, in whom peripheral blood stem cell from a genotypically identical sibling resulted in mixed chimerism.

The role of zinc sulfate in acute bronchiolitis in patients aged 2 to 23 months.

OBJECTIVE: Evaluating the effect of zinc sulfate in improving the clinical manifestations of acute bronchiolitis in children younger than 2 years. METHODS: This was a double blind pilot trial on 50 patients aged 2 to 23 months at Ghaem and Dr. Sheikh Hospitals in Mashhad from January 2008 to March 2009. Patients were randomly divided into two groups: a case group received oral zinc sulfate and to the control group was given placebo. FINDINGS: Mean age of case group was 168.0±108.6 days and control group 169.2±90.4 days (P=0.98) with male predominance in both groups. At first there was no statistically significant difference between the two groups in reducing the symptoms. But 24 hours after treating, improvement of some important manifestations including tachypnea, subcostal and intercostal retraction, wheezing and cyanosis revealed statistically significant difference in control group in comparison with case group (P=0.04). CONCLUSION: Zinc sulfate has no benefit in improving clinical manifestations of acute bronchiolitis.

Autoantibody profile in systemic sclerosis.

Systemic sclerosis is a generalized disorder of connective tissue clinically characterized by thickening and fibrosis of the skin and by distinctive forms of involvement of internal organs. One of the hallmarks of systemic sclerosis is the presence of serum autoantibodies against a variety of nuclear and cytoplasmic antigens. The primary purpose of this study was to identify the autoantibodies profile in the scleroderma sera and the secondary goal was to determine the correlation and discrepancy of autoantibody profile. Autoantibody profile was determined in 118 samples stored in the Advanced Diagnostic Laboratory at the University of Calgary. 78 sera were provided from Canadian and 40 sera were provided from Ukraine. We used the following techniques to identify autoantibodies profile in scleroderma patients: 1. Antinuclear antibody (ANA) by indirect immunofluorescence on human epithelial cell substrate 2. Detection and identification of specific autoantibodies by Innolia strip assay 3. Detection and identification of specific autoantibodies against extractable nuclear antigens. 111 out of 118 patients showed positive ANA results by indirect immunofluorescence and 7 patients had negative ANA results. Anti-ENA analyses by Inolia were positive in 84 patients, while by western blotting 81 patients showed positive results. In this study, we compared the results of anti-ENA antibody by Innolia with SLR technique. A significant correlation was found between anti-SC1-70 antibodies (P=0.000) and anti- RNP antibodies (P=0.001) and JO-1 antibodies (P=0.014). Thus, we may propose that SLR and Innolia techniques could be used for the detection of autoantibody in systemic sclerosis.

Analysis of human sera that are polyreactive in an addressable laser bead immunoassay.

BACKGROUND: Following the introduction of addressable laser bead immunoassays (ALBIA) into our clinical laboratory, it was noted that certain sera would exhibit reactivity to numerous antigens in the array. To further understand the nature of this reactivity, we analyzed the reactivity of sequential sera that were identified over a 1 year period. METHODS: Sera that demonstrated reactivity to 6 or more of the 8 antigens in an ALBIA kit (QuantaPlex 8: chromatin, Sm, RNP, Scl-70, ribosomal P protein, SS-A/Ro, SS-B/La, Jo-1) were tested for autoantibodies by indirect immunofluorescence (IIF) on HEp-2 cell substrates, for IgG, IgM and IgA rheumatoid factor, chromatin and ribosomal P protein by ELISA and by LINE immunoassay (LIA) and immunoblotting (IB). RESULTS: In one calendar year, 40/4096 (0.8%) sera analyzed in a routine clinical laboratory setting demonstrated reactivity to 6 or more antigens in the QuantaPlex 8 kits. There was no common IIF pattern that could be attributed to the polyreactive sera. There was no apparent correlation of polyreactivity with IIF titers, indeed, 4/40 (10%) sera had a negative ANA at the screening dilution of 1/160. When subjected to IB, LIA and ELISA, polyreactivity to three or more antigens was confirmed for 12/40 (30%) of sera while 8/40 (20%) had reactivity to 1-2 antigens and 20 (50%) did not react with any antigens in these assays. Overall agreement of positive or negative tests between the ALBIA and IB, LIA and ELISA was 75% for chromatin, 50% for SS-A, 27.5% for Sm, 25% for Rib-P, 22.5% for RNP, 20% for Scl-70, 15% for Jo-1 and 7.5% for SS-B. 17/40 (42.5%) had a positive IgM, IgG or IgA rheumatoid factor, and 12/40 (30%) had all three isotype rheumatoid factors. CONCLUSIONS: On average, the agreement between ALBIA and other assays in this study of polyreactive sera was 30%. Approximately, one-half of sera that demonstrate reactivity to multiple autoantigens in a commercial ALBIA were confirmed to have reactivity to at least one autoantigen in another diagnostic assay and 30% could be regarded as polyreactive. Other sera, some of which had rheumatoid factor, appeared to have high background binding without demonstrating specific binding to any of the cognate antigens.