Systemic disease in leukocytoclastic vasculitis: a focus on direct immunofluorescence findings

Abstract Background Direct immunofluorescence (DIF) panels are usually ordered for clinically suspected cutaneous vasculitis, but their positivity rate is variable, and their prognostic significance is not clear to date. Objective The study aims to investigate the systemic involvement rate in leukocytoclastic vasculitis (LCV) patients and the potential clinical and laboratory associations with systemic involvement, including DIF findings. Methods A retrospective study of patients with histopathologically proven cutaneous LCV examined in the dermatology department between 2013 and 2017 was performed. Results Of the 81 patients (mean age, 50.6 years), 42 (52%) were male. The mean time between the appearance of skin lesions and biopsy was 23.1 days, ranging from 2 to 180 days. DIF showed overall positivity of 90.1%, and C3 was the most frequent immunoreactant (82.7%). Any kind of extracutaneous involvement was present in 47 (58%) of patients, with renal involvement being the most frequent (53.1%), followed by articular (18.5%) and gastrointestinal (11.1%) involvement. The presence of renal disease was associated with the detection of IgG in the lesional skin (p = 0.017), and with the absence of IgM in the lesional skin (p = 0.032). There was a significant association between C3 deposition and joint involvement (p = 0.05). Study limitations This is a single-center study with a retrospective design. Conclusion DIF seems to be a useful ancillary diagnostic tool in the evaluation of cutaneous vasculitis, but the relationship between DIF findings and systemic involvement needs to be further elucidated due to contradictory data in the current literature.

Virus respiratorios y características clínico-epidemiológicas en los episodios de infección respiratoria aguda / Respiratory viruses and clinical- epidemiological characteristics in episodes of acute respiratory infection

RESUMEN Con el objetivo de determinar la frecuencia viral y las características clínico-epidemiológicas en los episodios de infección respiratoria aguda de pacientes del Instituto Nacional de Salud del Niño San Borja en Lima, se analizó la información de los episodios de pacientes que requirieron al menos una prueba de inmunofluorescencia directa desde el 1 de enero del 2017 al 31 de diciembre del 2018. Se identificaron 1153 episodios en 707 pacientes. La mediana de la edad fue de 1 año y el 55% fueron del sexo masculino. La frecuencia viral fue del 13,4%; el virus respiratorio sincitial se identificó en el 10,7% de los episodios. La frecuencia viral fue mayor en los menores de 1 año (16,2%); en aquellos con enfermedad congénita respiratoria (38,9%) y durante el otoño (24,2%). Los síntomas más comunes fueron tos (70,3%) y fiebre (53,4%); y los principales diagnósticos fueron neumonía viral (31,8%) y bronquiolitis (23,4%). Se concluye que la frecuencia viral respiratoria estuvo relacionada con la edad, estacionalidad y patología preexistente.

ABSTRACT The aim of the study was to determine the viral frequency and clinical-epidemiological characteristics in the episodes of acute respiratory infection in patients of the Instituto Nacional de Salud del Niño San Borja in Lima, the information of the episodes of patients who required at least one direct Immunofluorescence test from January 1, 2017 to December 31, 2018 was analyzed. 1153 episodes were identified in 707 patients. The median age was 1 year and 55% were male. The viral frequency was 13.4%; respiratory syncytial virus was identified in 10.7% of the episodies. The viral frequency was higher in children under 1 year of age (16.2%); in those with congenital disease respiratory (38.9%) and during the autumn (24.2%). The most common symptoms were cough (70.3%) and fever (53.4%); and the main diagnoses, viral pneumonia (31.8) and bronchiolitis (23.4%). It is concluded that the respiratory viral frequency was related to age, seasonality and pre-existing pathology.

Virus respiratorios y características clínico-epidemiológicas en los episodios de infección respiratoria aguda / Respiratory viruses and clinical- epidemiological characteristics in episodes of acute respiratory infection

RESUMEN Con el objetivo de determinar la frecuencia viral y las características clínico-epidemiológicas en los episodios de infección respiratoria aguda de pacientes del Instituto Nacional de Salud del Niño San Borja en Lima, se analizó la información de los episodios de pacientes que requirieron al menos una prueba de inmunofluorescencia directa desde el 1 de enero del 2017 al 31 de diciembre del 2018. Se identificaron 1153 episodios en 707 pacientes. La mediana de la edad fue de 1 año y el 55% fueron del sexo masculino. La frecuencia viral fue del 13,4%; el virus respiratorio sincitial se identificó en el 10,7% de los episodios. La frecuencia viral fue mayor en los menores de 1 año (16,2%); en aquellos con enfermedad congénita respiratoria (38,9%) y durante el otoño (24,2%). Los síntomas más comunes fueron tos (70,3%) y fiebre (53,4%); y los principales diagnósticos fueron neumonía viral (31,8%) y bronquiolitis (23,4%). Se concluye que la frecuencia viral respiratoria estuvo relacionada con la edad, estacionalidad y patología preexistente.

ABSTRACT The aim of the study was to determine the viral frequency and clinical-epidemiological characteristics in the episodes of acute respiratory infection in patients of the Instituto Nacional de Salud del Niño San Borja in Lima, the information of the episodes of patients who required at least one direct Immunofluorescence test from January 1, 2017 to December 31, 2018 was analyzed. 1153 episodes were identified in 707 patients. The median age was 1 year and 55% were male. The viral frequency was 13.4%; respiratory syncytial virus was identified in 10.7% of the episodies. The viral frequency was higher in children under 1 year of age (16.2%); in those with congenital disease respiratory (38.9%) and during the autumn (24.2%). The most common symptoms were cough (70.3%) and fever (53.4%); and the main diagnoses, viral pneumonia (31.8) and bronchiolitis (23.4%). It is concluded that the respiratory viral frequency was related to age, seasonality and pre-existing pathology.

Childhood pemphigus vulgaris is a challenging diagnosis

Pemphigus Vulgaris (PV) is an uncommon autoimmune and blistering mucocutaneous disease. Childhood Pemphigus Vulgaris (CPV) is a pediatric variant of PV, which affects children below 12 years, being very rare among children under 10 years of age. CPV has similar clinical, histological, and immunological features as seen in PV in adults. The mucocutaneous clinical presentation is the most common in both age groups. Vesicles and erosions arising from the disease usually cause pain. A few CPV cases have been reported in the literature. This study reports a case of an 8-year-old male patient with oral lesions since the age of 3 years, and the diagnosis of pemphigus was achieved only 2 years after the appearance of the initial lesions. CPV remains a rare disease, making the diagnosis of this clinical case a challenge due to its age of onset and clinical features presented by the patient. Therefore, dentists and physicians should know how to differentiate CPV from other bullous autoimmune diseases more common in childhood.

The importance of direct immunofluorescence in pemphigus herpetiformis diagnosis

Abstract Pemphigus herpetiformis is an autoimmune bullous disease, that combines clinical features of dermatitis herpetiformis and linear IgA bullous dermatosis and immunological characteristics of pemphigus, which makes this disease peculiar and this diagnosis rarely suspected in the first evaluation of the patient. The reported case is of a patient with clinically bullous disease similar to dermatitis herpetiformis, whose multiple biopsies were inconclusive, and only after direct immunofluorescence with a pemphigus pattern (intraepidermal intercellular pattern) the confirmation of the diagnosis was possible.

Update on the diagnosis and treatment of inherited epidermolysis bullosa / 中华皮肤科杂志

Inherited epidermolysis bullosa (EB) is a group of hereditary skin diseases characterized by increases in skin brittleness and being prone to skin blisters or erosions when the skin is exposed to slight friction or injury. At present, it is classified into four types, namely EB simplex, junctional EB, dystrophic EB and Kindler syndrome. An onion skinning approach is recommended for the diagnosis of inherited epedermolysis bullosa, in which, immunofluorescence mapping is performed firstly, and causative genetic loci are then determined through detection of relevant genes. In its treatment, skin care should be intensified, and medical or surgical treatment may be used to alleviate symptoms. Further development of cell?, protein?and gene?based therapies is expected to bring hope to patients via realization of timely prenatal diagnosis and avoidance of adverse consequences.

Rabia en Potos flavus identificados en el departamento de madre de dios, Perú / Rabies in Potos flavus identified in Madre de Dios, Peru

El Potos flavus es un mamífero nocturno que habita en bosques neotropicales desde Centroamérica hasta Sudamérica. Se realizó un estudio de cuatro casos de rabia en Potos flavus ocurridos desde abril de 2012 en el departamento de Madre de Dios en Perú, captados como parte de la vigilancia epidemiológica. Los análisis realizados en el laboratorio de referencia regional de Madre de Dios determinaron presencia de antígeno del virus de la rabia en tres de las muestras de tejido encefálico, dichos resultados fueron corroborados en el Laboratorio de Zoonosis Virales del Instituto Nacional de Salud del Perú mediante inmunofluorescencia directa, la tipificación no identificó ninguna de las variantes conocidas en murciélagos o en perros. La ocurrencia de cuatro casos de rabia en Potos flavus suma evidencias de la emergencia de un nuevo reservorio del virus de la rabia y que ha sido reportada previamente en el mismo departamento el año 2007.

The Potos flavus is a nocturnal mammal that lives in neotropical forests from Central America to South America. A study of four cases of rabies in Potos flavus was conducted; these occurred in April 2012 in the Madre de Dios region in Peru and were collected as part of epidemiological surveillance. The analysis performed in the regional reference laboratory of Madre de Dios determined the presence of the rabies virus antigen in three of the brain tissue samples. Results were verified in the Laboratory of Viral Zoonoses of the Peruvian National Institute of Health by direct immunofluorescence. The typification did not identify any of the known variants in bats or dogs. The occurrence of four cases of rabies in Potos flavus adds evidence of the emergence of a new reservoir of the rabies virus previously reported in the same region in 2007.

Comparison of different assays for detection of Chlamydia trachomatis / 国际检验医学杂志

Objective To investigate the sensitivity and specificity of different assays for detection of Chlamydia trachomatis . Methods Chlamydia trachomatis was determined in samples of cervical secretions from 213 patients with nongonococcal urethritis or genitourinary tract infection by direct immunofluorescence assay (DFA ) ,gold-immunochromatographic assay (ICA ) and real-time fluorescence quantitative polymerase chain reaction (FQ-PCR) .Results Comparing the positive rates of Chlamydia trachoma-tis detected by adopting the DAF ,FQ-PCR and the ICA methods showed the statistical difference (P<0 .05) .The sensitivity and specificity of Chlamydia trachomatis detected by DFA ,ICA and FQ-PCR were 95 .1% ,60 .2% ,97 .3% and 93 .2% ,99 .2% , 99 .3% ,respectively .The sensitivity of the DFA and FQ-PCR methods was higher than that of the ICA method ,difference was sta-tistically significant(P<0 .05) ,The specificity of the ICA and FQ-PCR methods was higher than that of the DFA method ,differ-ence was statistically significant (P<0 .05) .Conclusion The FQ-PCR method has higher sensitivity and higher specificity for the detection of Chlamydia trachomatis and can provide the reliable basis for clinically diagnosing the infection of Chlamydia trachom-atis .The primary medical units is suitable to adopt the ICA method for detecting Chlamydia trachomatis .

Sensitivities of seasonal and pandemic rapid antigen tests differentiated by the level of immunofluorescence for the detection of pandemic influenza A/H1N1 2009 virus

No abstract available.

A comparison between real-time quantitative fluorescence polymerase chain reaction assay and direct immunofluorescence assay for diagnosis of human metapneumovirus / 中华检验医学杂志

Objectives To evaluate the diagnostic value of real-time quantitative fluorescence polymerase chain reaction( Q-RT-PCR ) assay and immunofluorescence assay for diagnosis of hMPV. Methods Totally 1 283 children with acute respiratory infection admitted in Jiaxing Maternity and Child Health Care Hospital for treatment from November 2008 to May 2009 were recruited in this study. The hMPV positive stains were separated and sequenced in this area. The sequences between the local hMPV stains and Holland stains NLD00-1 were compared. The specific primers and fluorescent probe were designed according to the sequence of epidemic hMPV strain. The Taqman methodology was applied in Q-RT-PCR. Negative pressure suction was used to acquire nasopharyngeal secretions specimens. Both Q-RT-PCR and immunofluorescence with FITC labeled monoclonal antibody were used to analyze them, respectively. The McNemar, test was applied to analyze the correlation between the two methods. Results Totally 1 283 specimens were analyzed with Q-RT-PCR and immunofiuorescence simultaneously. Q-RT-PCR analysis showed there were 59 cases positive. Immunofluorescence analysis showed there were 55 cases positive. Fifty-two cases were positive in both assays. There were 7 cases positive in Q-RT-PCR assay but negative in immunofluorescence assay and 3 cases negative in Q-RT-PCR assay but positive in immunofluorescence assay. If Q-RT-PCR method was set as the golden standard, the sensitivity and specificity for immunofluorescence detection method were 88. 1%and 99. 8%, respectively. Positive predictive value and negative predictive value were 94. 5% and 99. 4%,respectively. There was no significant difference ( χ2= 0. 9, P ＞ 0. 05 ) by McNemar' test between the two methods. Conclusion The diagnostic value of immunofluorescence assay is close to Q-RT-PCR assay.

Desempeño de la prueba de inmunofluorescencia directa en el diagnóstico del virus Influenza A(H1N1) / Direct immunofluorescence assay performance in diagnosis of the Influenza A(H1N1) virus

El 25 de abril de 2009, a menos de un mes de la detección en México del primer humano con virus Influenza A(H1N1), la enfermedad ya se había propagado a más de 40 países superando los 10 000 casos notificados. Dada su naturaleza impredecible, este tipo de virus requiere métodos diagnósticos apropiados, confiables y seguros, pero que también estén al alcance de los laboratorios clínicos. Mediante el estudio de 291 muestras de pacientes con sospecha de infección por virus Influenza A(H1N1) en Neuquén, Argentina, el presente trabajo compara los dos métodos de diagnóstico utilizados simultáneamente: la prueba de inmunofluorescencia directa (DFA) y la de reacción en cadena de la polimerasa en tiempo real (RT-PCR). La DFA dio una sensibilidad de 44,4 por ciento, especificidad de 99,6 por ciento, valor predictivo positivo de 95,2 por ciento y valor predictivo negativo de 90,7 por ciento. Los resultados positivos de la metodología pueden considerarse verdaderos positivos. Un resultado negativo no excluye la presencia del virus y la muestra debe examinarse mediante RT-PCR. Del total de 291 muestras, 45 resultaron positivas por RT-PCR y 21 por DFA.

By 25 April 2009, less than one month after the first human with Influenza A(H1N1) virus was detected in Mexico, the disease had already spread to more than 40 countries, with over 10 000 cases reported. Due to its unpredictability, this type of virus requires appropriate, reliable, and safe diagnostic methods that are also accessible to clinical laboratories. Through the analysis of 291 samples taken from patients with suspected Influenza A(H1N1) virus infection in Neuquén, Argentina, this study compares the two diagnostic methods used simultaneously: direct immunofluorescence assay (DFA) and real-time polymerase chain reaction (RT-PCR). DFA had a sensitivity of 44.4 percent, a specificity of 99.6 percent, a positive predictive value of 95.2 percent, and a negative predictive value of 90.7 percent. Positive results obtained with this method can be considered true positives. A negative result does not rule out the presence of the virus. In this case, the sample should be examined by RT-PCR. Out of a total of 291 samples, there were 45 positive results with RT-PCR and 21 positive results with DFA.

Infecção por Chlamydia em pacientes com e sem lesões intra-epiteliais cervicais / Chlamydia infection in patients with and without cervical intra-epithelial lesions

OBJETIVOS: Determinar a freqüência de infecção por Chlamydia trachomatis em pacientes com e sem lesões intra-epiteliais cervicais atendidas em ambulatório especializado no Recife (2007), e sua associação com variáveis biológicas, demográficas, hábitos, características reprodutivas e clínico-ginecológicas. MÉTODOS: Realizou-se um estudo do tipo corte transversal, incluindo 70 mulheres (35 com alterações citológicas e 35 normais). Realizaram-se colposcopia, biópsia quando necessário e pesquisa para Chlamydia trachomatis por Imunofluorescência Direta. As variáveis analisadas foram idade, raça, procedência, escolaridade, estado civil, menarca, idade da primeira relação sexual, paridade, número de parceiros, corrimento, realização de citologia prévia, episódios de DST, eletrocauterização, método contraceptivo, antecedente familiar de câncer uterino, consumo alcoólico, tabagismo, drogas ilícitas e imunossupressoras, resultado da citologia e infecção cervical por Chlamydia trachomatis. Para determinação da força da associação, calculou-se a Razão de Prevalência (RP) e o intervalo de confiança 95 por cento, realizando-se análise multivariada para controle das variáveis potencialmente confundidoras. RESULTADOS: A freqüência de infecção por Chlamydia trachomatis foi significativamente maior em pacientes com alterações citológicas (80 por cento vs. 14,3 por cento), com uma RP de 5,60 (IC 95 por cento = 2,44 - 12,82). Analisando os fatores associados à infecção por Chlamydia , a única variável que persistiu significativamente associada após análise multivariada foi a história pregressa de DST (OR=63,47; IC 95 por cento = 13,93 - 289,09). CONCLUSÃO: A presença da Chlamydia trachomatis está associada às alterações citológicas da cérvice uterina, e a história pregressa de DST deve ser valorizada no tratamento e seguimento clínico destas pacientes.

OBJECTIVES: To determine the frequency of Chlamydia trachomatis infection in patients with and without intraepithelial cervical lesions attended at specialized ambulatory in Recife (2007) and its association with biologic and demographic variables, habits, reproductive, clinical and gynecologic characteristics. METHODS: A cross-sectional study was conducted including 70 women (35 with cytological cervical lesions and 35 with normal smears). Colposcopy and biopsy when necessary were performed and Chlamydia infection was determined by direct immunofluorescence. Analysis variables were age, race, precedence, schooling, marital status, menarche, age at first sexual intercourse, parity, number of sexual partners, vaginal discharge, previous Pap smear, STD episodes, cervical cauterization, contraceptive methods, familial antecedents of cervical cancer, alcohol intake, use of illicit and immunosuppressive drugs, Papanicolaou result and cervical Chlamydia l infection. To determine the strength of association prevalence ratio (PR) and its 95 percent confidence interval were calculated. Multivariate analysis was performed for controlling potentially confounding variables. RESULTS: the frequency of Chlamydia l infection was significantly greater in patients with intraepithelial cervical lesions (80.0 percent vs. 14.3 percent; PR = 5.60; 95 percent CI = 2,44 - 12,82). When the factors associated with Chlamydia l infection were analyzed, the only variable that remained significantly associated after multivariate analysis was previous episode of STD (OR=63,47; 95 percent CI = 13,93 - 289,09). CONCLUSION: Chlamydia trachomatis infection is associated with intraepithelial cervical lesions and history of STD should be considered for treatment and follow-up of these patients.

Chlamydia infection in patients with and without cervical intra-epithelial lesions tested by real-time PCR vs. direct immunofluorescence

This study compares the detection rates of Chlamydia trachomatis by two techniques, direct immunofluorescence (IMF) and real time polymerase chain reaction (PCR), in patients with and without intra-epithelial cervical lesions (SIL) in Recife. We conducted a transversal study involving 35 women with SIL and 35 without SIL attended at Ambulatório Especializado da Mulher, Recife, Brazil. They were tested for Chlamydia trachomatis using two techniques, direct IMF or real time PCR. The rates of Chlamydia trachomatis detection were compared and the association with intra-epithelial cervical lesions was determined using the chi-square test at a 5 percent level of significance. Concordance between the tests was evaluated using kappa. The global prevalence of Chlamydia infection was 47.1 percent by direct IMF and 58.6 percent by real time PCR. A significant association was observed between Chlamydia diagnosis and presence of intra-epithelial cervical lesions, with about 80 percent positive results by direct IMF and 77.1 percent by real time PCR. However, the detected rate of infection with Chlamydia trachomatis was significantly greater in patients without intra-epithelial cervical lesions tested by real time PCR (40 percent) when compared to direct IMF (14.3 percent). The concordance between the tests was weak, with a kappa coefficient of 0.4. Both real time PCR and direct IMF detected elevated rates of Chlamydia infection in patients with intra-epithelial cervical lesions (80 percent) but the tests were discordant when patients without cervical lesions were tested, possibly because sensitivity of real time PCR is greater.

Transient phosphorylation of tumor associated microtubule associated protein (TMAP)/cytoskeleton associated protein 2 (CKAP2) at Thr-596 during early phases of mitosis

Tumor associated microtubule associated protein (TMAP), also known as cytoskeleton associated protein 2 (CKAP2) is a mitotic spindle-associated protein whose expression is cell cycle-regulated and also frequently deregulated in cancer cells. Two monoclonal antibodies (mAbs) against TMAP/CKAP2 were produced: B-1-13 and D-12-3. Interestingly, the reactivity of mAb D-12-3 to TMAP/CKAP2 was markedly decreased specifically in mitotic cell lysate. The epitope mapping study showed that mAb D-12-3 recognizes the amino acid sequence between 569 and 625 and that phosphorylation at T596 completely abolishes the reactivity of the antibody, suggesting that the differential reactivity originates from the phosphorylation status at T596. Immunofluorescence staining showed that mAb D-12-3 fails to detect TMAP/CKAP2 in mitotic cells between prophase and metaphase, but the staining becomes evident again in anaphase, suggesting that phosphorylation at T596 occurs transiently during early phases of mitosis. These results suggest that the cellular functions of TMAP/CKAP2 might be regulated by timely phosphorylation and dephosphorylation during the course of mitosis.

Gene detection and clinical study of rhinovirus isolated from children with acute low respiratory tract infection in Shanghai / 中华传染病杂志

Objective To understand human rhinovirus (HRV) etiology of acute lower respiratory tract infection (ALRTI) in children in Shanghai area and establish a nested reverse transcription- polymerase chain reaction (nested RT-PCR) assay.Methods Three hundred and forty-two naso- pharyngeal secretion (NPS) samples from ALRTI cases who were hospitalized were collected during January 2005—December 2005.Nested RT-PCR techniques were used to detect HRV-specific RNA.The PCR products were sequenced and data of nucleotides were analyzed.The proportion of HRV infection in children with ALRTI,the distribution of gender,age and season,and clinical char- acteristics were also investigated.Results Forty-six (13.5%) of 342 samples were HRV positive detected by nested RT-PCR.The sequences of 15 positive samples shared high homology of 83%- 97% with HRV sequence in GenBank.Within the 15 positive samples,nucleotide homology varied from 64.4% to 98.4%,and the ratio of genetic variation was from 1.6% to 48.3%./00.These 15 ampli- cons attribute to the two branches of HRV cladogram.The sequences of 15 amplieons were highly varied,in which single nucleotide mutation and several nearby nueleotides mutations were found. Ribonucleotide deletion and insertion in the nucleotide sequence was also found.HRV positive sam- ples were detected in 33 boys and 13 girls,respectively.The ratio of infection cases between boys and girls was 2.5:1.Of 46 HRV infected cases,27 (58.7%) were less than 12 months of age and 38 (82.6%) were less than 3 years old.HRV infected ALRTI occured all the year round and peaked from March to May.Of the patients whose NPS samples were HRV positive detected by nested RT-PCR,45 patients were diagnosed with bronchopneumonia and 1 was diagnosed with asthmatic bronchitis.Fever of most patients was moderate.The peripheral blood leukocyte counts in thirty-nine (84.8%) patients were less than 10?10~9/L.Neutrophil percentages in thirty-seven (80.4%) patients were less than 0.50.C-reactive protein of thirty-six (78.3%) patients were less than 8 mg/L. All of these features were the characteristics of viral pneumonia.The complications were not common and conditions of most patients were not severe.All the children were cured.Conclusions This nes- ted RT-PCR technique is highly specific,rapid and convenient for the detection of HRV RNA in NPS of patients with ALRTI and the genome of HRV viruses is highly variable.The incidence of HRV infection predominates in children in Shanghai area.ALRTI of HRV is short of specificity and condi- tions of most patients are not severe and their prognoses are fine.