Search Results (19)

Background/Objectives: Anti-p200 pemphigoid is a rare and likely underdiagnosed autoimmune blistering disorder. Laminin γ1 and laminin β4 have been implicated as potential target antigens in its pathogenesis. Recently, a novel indirect immunofluorescence assay targeting anti-laminin β4 antibodies has been developed, demonstrating high sensitivity and specificity, and offering a valuable tool for improved diagnosis. Methods: Of the 451 patients, 21 were selected for further laboratory analysis based on medical records. Sera from 10 patients, which showed a positive direct immunofluorescence (DIF) result and negative results in multiplex enzyme-linked immunosorbent assays (ELISAs) and/or mosaic six-parameter indirect immunofluorescence (IIF) for various autoimmune bullous diseases, were tested for the presence of anti-laminin β4 antibodies. Additionally, sera from 11 patients with positive DIF and positive ELISA for antibodies against BP180 and/or BP230 were analyzed. Results: Among the 10 patients with positive DIF and negative ELISA and/or mosaic six-parameter IIF, 6 sera were positive for anti-laminin β4 antibodies. These patients presented with atypical clinical features. In contrast, all 11 sera from patients with both positive DIF and positive ELISA for BP180 and/or BP230 were negative for anti-laminin β4 antibodies. Conclusions: In patients with a positive DIF result but negative ELISA and/or mosaic six-parameter IIF findings, testing for anti-laminin β4 antibodies should be considered. Furthermore, in cases presenting with atypical clinical features—such as acral distribution of lesions, intense pruritus, or erythematous–edematous plaques—the possibility of anti-p200 pemphigoid should be included in the differential diagnosis. Full article

The rabies virus (genus Lyssavirus), is a deadly zoonotic agent affecting humans and animals. Although Mexico has been declared free of canine rabies (V1), sylvatic rabies persists. This study aimed to determine the prevalence of the virus in Desmodus rotundus and other non-hematophagous bat species in Oaxaca. The methodology comprised four stages: a literature review, data requests to the Servicio Nacional de Sanidad, Inocuidad y Calidad Agroalimentaria (SENASICA), fieldwork using mist nets across 15 municipalities in Oaxaca, and diagnosis via direct immunofluorescence at the Centro Nacional de Servicios de Diagnóstico en Salud Animal (CENASA). SENASICA reported 89 positive rabies cases (2014–2023) across six laboratories, with the majority (67.02%) attributed to the Oaxaca State Public Health Laboratory. Among the 194 bats analyzed (129 D. rotundus), only three tested positive for the virus, yielding a prevalence of 1.54%. Positive cases were exclusively identified in D. rotundus from San Lucas Ojitlán and The Heroic City of Tlaxiaco. This prevalence aligns with that of national studies, which ranges from 0.05% to 3%. These findings underscore the need to maintain epidemiological surveillance in wild and domestic fauna, alongside public awareness campaigns highlighting bats’ ecological importance for ecosystem conservation and the risks associated with their decline. Full article

Background and Objectives: Chlamydial infection is the most common asymptomatic infection worldwide. Despite all national programs, strategies and guidelines, chlamydial infection is still the leading infection worldwide, especially in young populations. We have tried to summarize the best diagnostic tools for its detection. Materials and Methods: In the study, 225 sexually active patients who were tested for chlamydial infection at the Institute of Public Health Kragujevac participated. Results: Combinations of direct immunofluorescence (DIF) and a rapid lateral immunochromatographic test (RT) and combinations of an RT and immunoglobulin G (IgG) do not improve diagnostic efficiency when compared to a rapid test that individually had the best parameters. In situations that require high specificity, the recommended combination is RT/IgA, which as a highly specific test has few false positive results, while the combinations of DIF + RT and RT + IgG, although showing a specificity of 100%, have low sensitivity (33.30%), due to which we prefer the RT/IgA combination. The combinations DIF + RT, DIF + RT + IgG and RT + IgG, although with low sensitivity, have the highest values of specificity, and the positive predictive value (PPV) and negative predictive value (NPV) show the highest values of the extended Youden index of 130.30% and the highest values of total diagnostic accuracy of 97.00%. Based on the results of the extended Youden index, taking into account PPV and NPV, the RT/IgA combination shows the highest value of 94.60%, as well as the highest value of total diagnostic accuracy of 93.00%. Conclusions: “Two or more positive tests” or “any test positive” did not improve the diagnostic efficiency compared to a single “rapid test”. Full article

Introduction: Epidermolysis bullosa acquisita (EBA) is a rare autoimmune disease causing subepithelial blistering due to autoantibodies against type VII collagen. While mechanobullous EBA predominantly affects adults, our report presents an exceedingly rare case in an 11-year-old football player. Case Report: The patient reported a one-year history of blistering and scarring on the knees and scrotum. The diagnosis was established with direct immunofluorescence (DIF), mosaic indirect immunofluorescence (IIF) showing IgG antibodies reacting with the dermal side of salt-split primate skin, and multiplex ELISA revealing an elevated level of IgG antibodies against type VII collagen. Treatment with a superpotent topical glucocorticosteroid and activity modifications improved his condition. Review: This case highlights the importance of considering EBA in differential diagnoses of pediatric blistering diseases and suggests that conservative management may be effective in mild cases. We also review clinical and laboratory considerations on the topic of childhood EBA. Conclusions: Further studies are essential to develop evidence-based guidelines for pediatric EBA. Full article

Background: Little is known about the relevance of BP180 ELISA for the clinical management of oral mucous membrane pemphigoid (OMMP). The aim of the present study was to investigate if the levels of anti-BP180 antibodies at diagnosis could be correlated with clinical severity and relapse. Methods: The present study included 44 OMMP patients with positive direct immunofluorescence (DIF). Circulating anti-BP180 IgG was measured using the same available ELISA kit (Euroimmun cut-off 20 U/mL). Clinical severity at diagnosis was measured using the oral disease severity score (ODSS). Only patients who achieved clinical remission (CR) were included in the analysis of variables related to relapse. Relapse was calculated as the interval between the date of the best type of clinical remission achieved and the date of relapse. Results: Values of BP180 > 20 U/mL significantly correlated with higher ODSSs in both univariate (p < 0.05) and multivariate analyses (p < 0.05). Among 39/44 patients who achieved CR, 17/39 relapsed. Kaplan–Meier analysis revealed that patients with BP180 > 20 U/mL displayed worse clinical behavior in terms of relapse (p < 0.05). Conclusion: BP180 ELISA at diagnosis appears to be a useful parameter to stratify OMMP patients with more severe disease and worse clinical outcomes after clinical remission. Full article

Senear–Usher syndrome, or pemphigus erythematosus (PE), is a rare autoimmune disorder characterized by the coexistence of features from both lupus erythematosus (LE) and pemphigus foliaceus (PF). We describe a 41-year-old patient initially diagnosed with cutaneous and then systemic lupus erythematosus (SLE), who after a few years developed new skin lesions: erythematous and erosive eruptions partially covered by crusts located on the trunk and flaccid blisters on the extremities. Direct immunofluorescence of perilesional skin revealed deposits of IgG in the intercellular space of the epidermis and granular deposits of C3 at the dermo–epidermal junction. Additional testing, revealing autoantibodies against the intercellular space of the epidermis, and direct immunofluorescence (DIF) examination allowed a diagnosis of pemphigus vulgaris coexisting with lupus. Further, DIF study revealed granular deposits of immunoglobulin G (IgG) in the intercellular spaces of the epidermis and granular deposits of the C3 along the basement membrane. Clinical appearance led to suspicion of Senear–Usher syndrome. in this patient. This case report explores the diagnostic challenges posed by the patient’s overlapping symptoms and immunological findings, suggesting an infrequent manifestation of Senear–Usher syndrome or a combination of SLE and pemphigus vulgaris. The case highlights the complexity of chronic inflammatory skin diseases and the need for tailored treatment approaches in such cases. Despite temporary improvement, the patient experienced relapses. We performed a descriptive literature review of the case reports of PE published in the last 24 years and prepared a summary of the characteristics, emphasizing the importance of proper recognition, clinical features, and treatment of this uncommon syndrome. Full article

Background/Objectives: Pemphigus comprises a diverse group of disorders within the autoimmune bullous dermatoses (AIBDs) spectrum. Among these, pemphigus vulgaris (PV) and pemphigus foliaceus (PF) are the most commonly encountered variants. Despite its rarity, this condition can pose a life-threatening risk. We aimed to assess clinical characteristics, comorbidities, medication, as well as the treatment of various types of pemphigus in pemphigus patients. Methods: We gathered data from 69 patients treated in the Department of Dermatology in the years 2016–2023. The investigation included sex, age at diagnosis, type of pemphigus, comorbidities and medications, presence of neoplasms and treatment of pemphigus, as well as enzyme-linked immunosorbent assay (ELISA) and direct immunofluorescence (DIF) results. The data were statistically analyzed with the p-value set at 0.05. Results: The study group comprised 69 patients, including 41 women and 28 men. The mean age at diagnosis was 56.89 years +/− 15.42 years. A total of 79.31% of the patients were diagnosed with PV and the following 26.09% with PF. The most common comorbidities were arterial hypertension, hypercholesterolemia, and diabetes mellitus. The dominant treatment regimen was the systemic use of glucocorticosteroids (GCSs; 90% and 94% of PV and PF patients, respectively). More than half of the patients received at least one GCS-sparing treatment, including dapsone and rituximab. We observed a significantly frequent presence of IgG deposits in DIF in patients with PF (p = 0.0217) and a subsequent correlation between the concurrent presence of IgG deposits in DIF and anti-DSG1 antibodies in ELISA testing (p = 0.0469). The combination of IgG, IgG1, IgG4, and C3 deposits was more often existent in PF patients (p = 0.0054) and the combination of IgG4 and C3 deposits in PV patients (p = 0.0339). We also found a positive correlation between the level of anti-DSG1 antibodies and the age at diagnosis (p = 0.0298). Conclusions: Patients with pemphigus are very often diagnosed with significant comorbidities and take diverse medication, which shows that the treatment of pemphigus should follow a multidisciplinary approach. Accurate analysis of the clinical condition of the patients, as well as the results of the ELISA panel or DIF, is crucial for a successful diagnostic and therapeutic process. Full article

Objectives: Our study aimed to establish the basic reliability parameters of direct immunofluorescence test results in patients with oral lichen planus. Methods: We conducted an evaluation of individual antibody classes in the DIF and ELISA (BP180 antigen), comparing these results with the classical histopathological (HP) examination in a group of patients treated within the standard healthcare in our clinic. Results: Among 66 participants with oral changes indicative of LP, only 50% received histopathological confirmation of the LP diagnosis. Among those with a DIF profile entirely typical for LP (C3+, F+), 57.1% had a positive HP result. Fibrinogen deposits were identified in 42.4% and 36.4% of individuals with positive HP results for F1 and F2, respectively; 78.8% of patients with negative HP and 57.6% with positive HP exhibited no fibrinogen deposits. Simultaneous positivity for F1 and F2 occurred in all cases where F1 was positive. HP confirmed positive DIF for C3 in 50% of cases. Fibrinogen deposits demonstrated the highest diagnostic accuracy (61%). Sensitivity and specificity for fibrinogen deposits were 36% and 42% for F1 and 79% and 82% for F2. The positive predictive values were 67% for F1 and 67% for F2, while the negative predictive values were 58% for F1 and 56% for F2. Overall diagnostic accuracy was reported at 61% for F1 and 59% for F2. Conclusions: Our data indicate the complementarity of HP and immunological test results and the necessity of using both methods together in cases of doubt. Full article

The diagnosis of oral lichen planus (OLP) poses many challenges due to its nonspecific clinical symptoms and histopathological features. Therefore, the diagnostic process should include a thorough clinical history, immunological tests, and histopathology. Our study aimed to enhance the diagnostic accuracy of OLP by integrating direct immunofluorescence (DIF) results with clinical data to develop a multivariate predictive model based on the Artificial Neural Network. Eighty patients were assessed using DIF for various markers (immunoglobulins of classes G, A, and M; complement 3; fibrinogen type 1 and 2) and clinical characteristics such as age, gender, and lesion location. Statistical analysis was performed using machine learning techniques in Statistica 13. The following variables were assessed: gender, age on the day of lesion onset, results of direct immunofluorescence, location of white patches, locations of erosions, treatment history, medications and dietary supplement intake, dental status, smoking status, flossing, and using mouthwash. Four statistically significant variables were selected for machine learning after the initial assessment. The final predictive model, based on neural networks, achieved 85% in the testing sample and 71% accuracy in the validation sample. Significant predictors included stress at onset, white patches under the tongue, and erosions on the mandibular gingiva. In conclusion, while the model shows promise, larger datasets and more comprehensive variables are needed to improve diagnostic accuracy for OLP, highlighting the need for further research and collaborative data collection efforts. Full article

Improving the management of children with asthma associated with mycoplasma infection is important. Aim: To study the duration of the persistence of antigens, and DNA in a free state, in the structures of circulating immune complexes (CICs) and living cells of Mycoplasma pneumoniae (Mpn) and Mycoplasma hominis (Mh) in children with asthma. In total, 205 children with asthma from 1 to 14 years were observed. The reaction of aggregate-hemagglutination (AHAA), the direct immunofluorescence reaction (DIF), the reaction of the polymerase chain reaction (PCR), and the culture method were used. In addition, 47 children were re-examined 1.5 months after the treatment of mycoplasma infection with azithromycin. The number of samples positive for antigens and DNA in the free state and in the structures of CICs significantly decreased. Then, 50 blood serum samples containing Mh antigens, and 50 samples containing Mpn antigens were analyzed by culture method. Mh was isolated in 21 (65.5%) of 32 samples containing DNA. Mpn was isolated from antigen-positive samples in nine cases. The presented data indicate the long-term persistence of antigens, and DNA of mycoplasma cells in the free state, in the structure of CICs, as well as in the form of “microcolonies”. A high level of CICs can be used to predict the course of the disease and the response to therapy. Full article

Mucous membrane pemphigoid (MMP), also known as cicatricial pemphigoid (CP), is a heterogeneous group of subepidermal blistering diseases that affect the mucous membranes, most frequently in the eye and oral cavity. MMP is often unrecognized or misdiagnosed in its early stages due to its rarity and nonspecific presentation. We present the case of a 69-year-old female in which MMP of the vulva was not initially suspected. The first biopsy, from lesional tissue for routine histology, revealed fibrosis, late-stage granulation tissue, and nonspecific findings. A second biopsy, from perilesional tissue for direct immunofluorescence (DIF), revealed DIF findings typical of MMP. Scrutiny of both the first and second biopsies revealed a subtle but telling histologic feature: subepithelial clefts along adnexae in the context of a scarring process with neutrophils and eosinophils, which can be an important clue to MMP. This histologic clue has been previously described; reinforcing its importance may prove useful for future cases, especially those for which DIF is not feasible. Our case demonstrates the protean presentations of MMP, the need for persistence in sampling unusual cases, and the relevance of inconspicuous histologic features. The report highlights this underrecognized yet potentially decisive histologic clue to MMP, reviews current biopsy guidelines when MMP is suspected, and delineates the clinical and morphological features of vulvar MMP. Full article

Dermatitis herpetiformis (Duhring’s disease, DH) is a chronic blistering cutaneous condition with pruritic polymorphic lesions, consisting of vesicles, papules or nodules and erythema, found predominantly on the extensor surfaces of the limbs, buttocks, and neck. Diagnosis is based on characteristic clinical and immunopathological findings. Oral manifestations of DH have rarely been described. The aim of the study was to evaluate IgA, IgG, IgM and C3 complement deposits in the oral mucosa in DH patients. Direct immunofluorescence (DIF) was performed on the oral mucosa specimens collected from 10 DH patients. Biopsy was taken in a local anesthesia from perilesional site from the buccal mucosa and then preserved in a standard procedure using polyclonal rabbit IgG, IgA, IgM and C3 antibodies. Granular IgA and C3 deposits were found in 6 patients (60%), and in 3 subjects (30%) the result was indeterminate. Significant fluorescence of the deposits along the basement membrane was observed in 2 patients, moderate fluorescence in 3 patients, and in 4 cases the result was indeterminate. C3 deposits were found in 5 subjects (50%), 3 of them being moderate and 2 indeterminate. No IgM and IgG deposits were detected in the collected buccal mucosa specimens. Full article

Introduction: Autoimmune bullous dermatoses (ABD) represent a heterogeneous group of blistering disorders that may be debilitating with high morbidity. Clinical, histological, and direct immunofluorescence (DIF) studies are essential in establishing an accurate diagnosis of ABD, which is essential for its clinical management. Our study objective was to perform a systematic evaluation of ABD cases in a patient population at an academic medical center in Ho Chi Minh City, Vietnam, and determine the degree of concordance of clinical, histological, and DIF findings in ABD. Methodology: A systematic retrospective cross-sectional study was performed on 92 patients diagnosed with ABD by clinical, histological, and DIF studies at the University of Medicine and Pharmacy in Ho Chi Minh City, Vietnam, between September 2019 and September 2021. The clinical histories, H and E stained tissue sections, and DIF stains were evaluated by pathologists at the University of Medicine and Pharmacy. Results: ABD was evaluated as a whole and subdivided into an intraepidermal blister subgroup and a subepidermal blister subgroup. The analysis of paired diagnostic methods (clinical, histological, and DIF) for concordance with the final diagnosis was performed and showed that there were no statistically significant differences between the paired methods (McNemar’s test, p > 0.05). There was moderate concordance between the clinical, histological, and DIF diagnoses among all ABD cases (Brennan-Prediger coefficient Kappa test, κ_BP = 0.522, CI = 0.95). In the intraepidermal blister subgroup, the diagnostic accuracies of the histology and DIF stains were comparable to each other, and both were more accurate than a clinical diagnosis alone. In the subepidermal blister subgroup, there was no statistically significant difference in each pair of the three diagnostic methods (clinical, histological, and DIF) (McNemar’s test, p > 0.05). The concordance between the clinical, histological, and DIF diagnoses was high for the intraepidermal blister subgroup (Kappa test, κ_BP = 0.758, CI = 0.95). However, the concordance between the clinical, histological, and DIF diagnoses was slight for the subepidermal blister subgroup (Kappa test, κ_BP = 0.171, CI = 0.95). Conclusion: Histological evaluation is highly accurate in the diagnosis of the intraepidermal blister subgroup, but it is not as accurate in the diagnosis of the subepidermal blister subgroup in the Vietnamese patient cohort in which clinical, histological, and DIF studies were performed. DIF stains are a crucial diagnostic tool for ABD in this patient population. Full article

Chronic ulcerative stomatitis (CUS) is a rarely reported disease affecting the oral cavity, most often affecting middle-aged Caucasian females. The aim of the present study is to present the diagnosis, differentiation, and interdisciplinary treatment of this rare disease. CUS is characterized by the presence of an oral erosive or ulcerative lesion. The autoimmune pathogenesis of CUS includes affecting the antigen’s activity by DNA-breaking and protein-hydrolyzing enzymes. The stratified epithelium-specific antinuclear antibodies (SES-ANA) are associated with CUS development. Clinically, the lesions presented in oral mucosa might resemble an erosive form of oral lichen planus, whereas gingival lesions seem to be similar to desquamative gingivitis related to dermatological diseases manifested in the oral cavity. Patients often report subjective symptoms related to oral mucosa and general symptoms. Histopathological presentation of CUS is often non-specific and includes sub-epithelial separation from underlying connective tissue, atrophic epithelium, and inflammatory infiltrate with an increased number of plasma cells and lymphocytes. Direct immunofluorescence (DIF) might be used in CUS diagnostics. CUS generally remains nonsusceptible to corticosteroid treatments; however, antimalarial drugs and calcineurin inhibitors are more effective. Further research should be conducted in order to implement a diagnostic protocol and observe the long-term results of CUS management. Full article

Leishmania major cutaneous leishmaniasis (CL) lesions are characterized by an intense process of parasite destruction and antigen processing that could limit microscopic amastigote detection. The aim of our study was to develop a direct immunofluorescence (DIF) assay for in situ visualization of L. major antigens and access its reliability in the routine diagnosis of CL. The developed DIF assay used IgG polyclonal antibodies produced in rabbits by intravenous injections of live L. major metacyclic promastigotes chemically coupled to fluorescein isothiocyanate. Applied to L. major infected RAW macrophages, corresponding macrophage-derived amastigotes and dermal scrapings from CL lesions, the immunofluorescence assay stained specifically Leishmania amastigotes and showed a diffuse Leishmania antigen deposit into cytoplasm of phagocytic cells. Reliability of DIF in CL diagnosis was assessed on 101 methanol-fixed dermal smears from 59 positive and 42 negative CL lesions diagnosed by direct microscopy and/or kDNA real-time PCR. Sensitivity and specificity of DIF was 98.3% and 100%, respectively, being more sensitive than microscopy (p < 0.001) and as sensitive as ITS1-PCR. ITS1-PCR-RFLP allowed Leishmania species identification in 56 out of the 58 DIF-positive smears, identifying 52 L. major, two L. infantum and two L. tropica cases, which indicates antigenic cross-reactivity between Leishmania species. Full article