Background/Aim: Autoimmune bullous diseases are characterised by the production of autoantibodies to epidermal or subepidermal adhesive proteins. The aim of this study was to determine the ...relationship between age and sex of patients with the results of indirect immunofluorescence test in patients with newly diagnosed bullous dermatoses. Methods: The investigation presents a retrospective study of newly diagnosed patients with autoimmune bullous diseases at the Clinic for Skin and Venereal Diseases of the University Clinical Centre in Banja Luka in the period 2016-2021. In addition to demographic data, the results of an indirect immunofluorescence test in two titres (≥ 1:10 and ≥ 1:100) were analysed. Results: In this study, almost the same number of patients with pemphigus (45.2 %) and pemphigoid (54.8 %) was found. There were more women than men in the total sample (p = 0.049). The average age of subjects with pemphigoid was higher than that of patients with pemphigus (p = 0.001). 48.2 % of patients with pemphigus and 51.8 % of patients with pemphigoid had a positive indirect immunofluorescence test. A positive test for epidermal intercellular substance in both sexes at a titre ≥ 1:100 is higher than a titre ≥ 1:10 (p = 0.029). Patients with autoantibody titres ≥ 1:100 to desmoglein-1 were statistically significantly older than patients with titres ≥ 1:10 (p = 0.047). Conclusion: Number of patients with pemphigus and pemphigoid were similar, with no difference in sex distribution between the two groups of patients, but patients with pemphigoid were older than patients with pemphigus. The difference between high and low autoantibody titres in both sexes was found only in the group of pemphigus on epidermal intercellular substance and desmoglein-1.
Context: A search for a cause for membranous nephropathy (MN) is crucial to determine its treatment and management. Primary MN was a diagnosis of exclusion until the discovery of the target antigen, ...phospholipase A2 receptor (PLA2R). Lack of published data from the Indian population prompted this prospective study to determine the sensitivity and specificity of circulating anti-PLA2R antibodies in MN patients by using cell-based indirect immunofluorescence test (IIFT) and correlating with clinical-histopathology features and response to treatment. Settings and Design: This was a cross-sectional prospective study. Materials and Methods: MN cases (n = 34) diagnosed by renal biopsy and IIFT were evaluated along with 10 controls for serum anti-PLA2R antibodies using IIFT on biochip containing HEK 293 cell lines transfected with cDNA coded for PLA2R in this cross-sectional prospective study and simultaneously investigated to find the cause for MN. Positive cases treated with the Ponticelli regimen were followed up for 6 months with repeat testing for PLA2R. Statistics were performed using Statistical Package for Social Sciences version 18 (IBM). P < 0.05 considered significant. Statistical parameters were analyzed using the Chi-square test. Results: Anti-PLA2R antibodies-positive MN (primary MN) cases (n = 20) had higher 24-h proteinuria (10.09 ± 2.46 g) with 25% cases showing mesangial hypercellularity and basement membrane thickening in all (100%), while 50% of secondary MN cases showed mesangial hypercellularity with 7.17 ± 3.8 g of proteinuria. The sensitivity, specificity, and accuracy rate of anti-PLA2R antibodies for a diagnosis of primary MN were 70%, 100%, and 82%, respectively. Conclusion: Anti-PLA2R antibody in serum is a good reliable noninvasive diagnostic biomarker for primary MN and for monitoring its disease activity.
As in other viral infections, anti-nuclear antibodies (ANAs) are observed in SARS-CoV-2 infection. We investigated the presence of autoantibodies in acute COVID-19 and the association with early ...laboratory findings.
We examined 50 sera (>18 years, 25 Female) from patients with acute COVID-19. ANAs (HEp-20-10 liver biochip), anti-neutrophil cytoplasmic antibody (ANCA, Europlus Granulocyte Mosaic 32) and anti-double stranded DNA were investigated with product of Euroimmune AG (Luebeck, Germany) by indirect immunofluorescence (IIF) method. Also, antibody against cyclic citrullinated peptide (anti-CCP) was examined by a chemiluminisens assay (Euroimmun AG, Luebeck, Germany). Samples from 50 blood bank donors collected before the COVID-19 pandemic were used as controls.
The IIF-ANA test was positive in 18% (N = 9/50) of the patients. The median time of sample collection was 7 days (range: 1–28 days) after diagnosis. ANA was positive in only one (2%) control sample. Five (55.5%) patients were ANA positive with a strong titer (3+). There was no relationship between antibody titration and time of sample collection (p = 0,55). Anti-CCP was detected in a nucleolar (3+) positive patient (2%). ANA was detected in 14.28% (N = 1/7, rods-rings (±), p = 0,78) of patients in the intensive care unit(ICU). Patients treated in the clinic have more and higher titers of ANA, mostly in nucleolar patterns, than ICU patients.
The variety of antibodies detected in acute COVID-19 and the uncertainty of how long they persist can lead to confusion, especially in the diagnosis of systemic autoimmune rheumatic diseases for IIF-ANA testing in immunology laboratories. Improvements in cell lines and methods will facilitate the diagnostic process.
•ANAs were in higher titers and mainly in nucleolar patterns in acute COVID-19 in the IIF assay.•ANAs could be observed together with vasculitis-associated antibodies and/or extractable ANA patterns in acute COVID-19.•ANA titers were higher in patients monitored in the clinic than in the intensive care unit.•The variability and uncertainty in antibody response in COVID-19 may pose a challenge in diagnosing SARD for IIF testing.
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GEOZS, IJS, IMTLJ, KILJ, KISLJ, NLZOH, NUK, OILJ, PNG, SAZU, SBCE, SBJE, UILJ, UL, UM, UPCLJ, UPUK, ZAGLJ, ZRSKP
Currently, duck Tembusu virus (DTMUV), an emerging avian pathogenic flavivirus, is widely spread and becomes endemic in duck populations in Asia, causing significant economic losses in the duck ...producing industry. To early detection and control of DTMUV, the well‐validated diagnostic tests for efficient detection of DTMUV infection in ducks are needed. In this study, we validated and compared hemagglutination inhibition (HI) and indirect immunofluorescence (IFA) tests for identifying antibodies against DTMUV in duck serum samples. Our results demonstrated that HI and IFA tests can both be used to detect antibodies against DTMUV in duck serum samples with high sensitivity (100%), specificity (>87%) and overall agreement with the gold standard serum neutralization (SN) test (>90%). Additionally, DTMUV‐specific antibody titres determined by HI and IFA tests correlated well with the neutralizing antibody titres obtained by SN test. No cross‐reactivity against common duck viruses and other flaviviruses was observed in both tests. It is interesting to note that HI test had higher diagnostic specificity and exhibited a stronger positive correlation with SN test than IFA test. Evaluating the performance of HI and IFA tests with experimental and field serum samples revealed that both tests showed comparable performance with SN test in terms of antibody kinetic and detection rate. Collectively, these findings support the use of both tests, particularly HI test, as the alternative to SN test for measuring the antibody responses against DTMUV in ducks. These tests could be the suitable choices for DTMUV diagnosis, epidemiological study and vaccine efficacy evaluation.
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BFBNIB, DOBA, FZAB, GIS, IJS, IZUM, KILJ, NLZOH, NUK, OILJ, PILJ, PNG, SAZU, SBCE, SBMB, UILJ, UKNU, UL, UM, UPUK
Feline leishmaniosis (FeL) is an emerging vector-borne feline disease, with increasing numbers of cases reported and studies performed internationally. This study aimed to update the epidemiological ...status for FeL in stray cats in Milan, northern Italy; compare these results with previous studies in Northern Italy; and report clinicopathologic findings and coinfections in cats infected with
spp. A total of 117 cats were tested for
and retrovirus infection, hematological, and biochemical parameters. Demographic and clinical data were collected and FeL affected cats screened for selected coinfections. Overall, 10/117 (8.6%) cats tested positive for
: in five cats
DNA was found in popliteal lymph nodes and five were IFAT seropositive at titers from 1:80 to 1:160. Infected cats were concentrated in a specific area of Milan (
= 0.0154). No specific clinicopathologic abnormalities or retroviral infections were significantly linked to the infection, other than hypergammaglobulinemia (
= 0.0127). Seroreactivity to
,
and
was found in some infected cats. A high prevalence of FeL was found in a non-endemic area of northern Italy and future studies should continually monitor this data to understand whether these cases are imported or if
vectors are present in this area.
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DOBA, IZUM, KILJ, NUK, PILJ, PNG, SAZU, UILJ, UKNU, UL, UM, UPUK
Objectives
The aims of this study were to investigate the prevalence of Leishmania species infection in cats in Northern Italy and to evaluate the associations between infection and signalment and ...clinicopathological data.
Methods
The study was carried out in a veterinary university hospital from June to November 2017. Blood, urine, conjunctival swabs and hair were collected from all randomly selected cats. Leishmania species infection was evaluated using the indirect fluorescent antibody test (IFAT), setting a cut-off value of 1:80, and using real-time PCR on blood, conjunctival and hair samples. A complete blood count, serum chemistry profile, serum electrophoresis and urinalysis were also carried out. The cats were grouped on the basis of the results of the diagnostic criteria adopted in positive, negative and unconfirmed Leishmania cases. Non-parametric variables and continuous data were compared among the study groups using the χ2 test and the Mann–Whitney U-test, respectively.
Results
One hundred and fifty-two cats were included. Nineteen of the 152 (12.5%) cats were positive (18/152 11.8% showed an IFAT titre of ⩾1:80 and 1/152 0.7% was real-time PCR-positive from a hair sample); 106/152 (69.7%) cats were negative; and 27/152 (17.8%) cats were unconfirmed for Leishmania species. Total proteins, beta2-globulin and gamma-globulin were significantly increased in the positive Leishmania group compared with the negative group.
Conclusions and relevance
The results of the present study demonstrated the spread of Leishmania infantum infection in cats in Northern Italy. Hyperproteinaemia and hypergammaglobulinaemia appeared to be significant clinicopathological abnormalities in this population of cats with L infantum infection.
Climate change, increased urbanization and international travel have facilitated the spread of mosquito vectors and the viral species they carry. Zika virus (ZIKV) is currently spreading in the ...Americas, while dengue virus (DENV) and chikungunya virus (CHIKV) have already become firmly established in most tropical and also many non-tropical regions. ZIKV, DENV and CHIKV overlap in their endemic areas and cause similar clinical symptoms, especially in the initial stages of infection. Infections with each of these viruses can lead to severe complications, and co-infections have been reported. Therefore, laboratory analyses play an important role in differential diagnostics. A timely and accurate diagnosis is crucial for patient management, prevention of unnecessary therapies, rapid adoption of vector control measures, and collection of epidemiological data.There are two pillars to diagnosis: direct pathogen detection and the determination of specific antibodies. Serological tests provide a longer diagnostic window than direct methods, and are suitable for diagnosing acute and past infections, for disease surveillance and for vaccination monitoring. ELISA and indirect immunofluorescence test (IIFT) systems based on optimized antigens enable sensitive and specific detection of antibodies against ZIKV, DENV and CHIKV in patient serum or plasma. In recent years, Euroimmun (Lübeck, Germany) has developed numerous test systems for the serological diagnosis of (re-)emerging diseases, including a very sensitive and specific anti-ZIKV ELISA.
Anti-dsDNA antibody is a specific antibody in systemic lupus erythematosus (SLE). Indirect immunofluorescence test (IIFT) is a highly specific method in detecting anti-dsDNA antibody. The application ...of automated system has gained better consistency than manual operation. This study detected anti-dsDNA antibodies using EUROPattern Computer-aided immunofluorescence microscopy (EPA), and evaluated the performance of the automated system.
The sera of 96 patients with suspected SLE and 102 control patients were examined using IIFT. The consistency between the EPA and manual reading was analyzed.
Analysis of 198 samples showed that the overall consistency of the negative/positive results between the EPA and manual reading was 94.95%. Based on the manual reading results, the sensitivity and specificity of EPA were 95.70% and 94.29%, respectively. The analysis of 57 samples with non-specific fluorescence showed that the overall consistency of the negative/positive results was 96.49%. The analysis of the antibody titer of 89 positive samples showed that the consistency between the EPA and manual reading was 97.75%.
EPA was consistent with the manual reading with regard to qualitative reading and antibody titer. With low-exposure function, EPA could read samples with non-specific fluorescence. EPA was superior to manual reading in automation and standardization.
•The consistency between automatic (EPA) and manual reading was good for negative/positive results and antibody titer.•The new function of low-exposure imaging improved the consistency of samples with non-specific fluorescence.•The automated system(EPA) will promote the automation and standardization of IIFT.
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GEOZS, IJS, IMTLJ, KILJ, KISLJ, NLZOH, NUK, OILJ, PNG, SAZU, SBCE, SBJE, UILJ, UL, UM, UPCLJ, UPUK, ZAGLJ, ZRSKP
Asymptomatic cats often harbor pathogens, some of which have not been largely investigated in feline populations. The aim of this study was to evaluate the occurrence of antibodies against Rickettsia ...conorii, Rickettsia felis, Rickettsia typhi, Neospora caninum, Bartonella henselae and Toxoplasma gondii in cats from Tuscany. Ninety-five blood serum samples, previously collected, were analyzed by indirect immunofluorescence assay. Fifty-six (58.94%) cats had antibodies to at least one investigated pathogen: 28 (29.47%) cats were positive for B. henselae, 17 (17.89%) for R. felis, 14 (14.73%) for R. conorii, 14 (14.73%) for T. gondii, 2 (2.1%) for N. caninum. No cats were positive for R. typhi. Positive reactions to two or more pathogens were detected in 18 (18.94%) cats. The occurrence of antibodies against these microorganisms suggests that cats, even though asymptomatic, may be infected by pathogens, often zoonotic, and thus may be a source of infections for other animals and humans.
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DOBA, IZUM, KILJ, NUK, PILJ, PNG, SAZU, UILJ, UKNU, UL, UM, UPUK
Background: Antinuclear antibody (ANA) testing is the first test ordered in any patient suspected of autoimmune diseases. Often ANA indirect immunofluorescence (IIF) screening tests have high ...sensitivity because of high background, which can lead to erroneous clinical diagnosis and management when ordered unnecessarily. The following study was undertaken to envisage the clinical importance of ANA-positive test results among cases presenting to a tertiary neuro-care center. Methods: Present study is a secondary data analysis of 166 ANA IIF positive samples. Serum samples were further subjected to specific antigen detection by ANA immunoblot assay. The final diagnosis was reviewed in the context of descriptive data as to why testing was ordered. Results: The overall positivity rate of ANA for 3 years was 2.12%. The peripheral nervous system was most commonly involved accounting for 56 (33.73%) cases followed by the central nervous system in 40 (24.09%) cases and others. The predominant ANA pattern observed was speckled in 73 (43.9%) cases. On ANA profiling, reactivity to Ro52 (35 40.22%) was most common. Females were predominantly (70.5%) affected than males (29.5%). There was no association found in ANA pattern with age, sex, duration of illness, and the clinical disorder. Conclusions: ANA positivity is sporadically associated with nervous system autoimmune disorders. Screening tests like ANA can be judiciously employed for meaningful clinical interpretation of neurological diseases.
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