Comparative study of direct and indirect immunofluorescence for diagnosis of canine pemphigus foliaceus / Estudo comparativo da imunofluorescência direta e indireta para o diagnóstico do pênfigo foliáceo canino

Pemphigus foliaceus (PF) is the most common autoimmune skin disease in dogs. It is characterized by pustules, erosions, and crusts which occur due to the presence of autoantibodies that target intercellular adhesion. Histopathological examination is considered the gold standard pattern in the diagnosis, but may sometimes be inconclusive, especially when the characteristic findings are not identified. New diagnostic tests are continuously being developed and immunofluorescence assays, could be a valuable alternative diagnostic tool. This study aimed to evaluate the applicability of direct and indirect immunofluorescence (DIF and IIF) tests for the diagnosis of canine PF. Twenty eight dogs were divided into two groups: Group I with 14 dogs with PF and Group II (control) with 14 dogs with Superficial pyoderma (differential diagnoses of PF). All animals were submitted to skin biopsy to histopathological and DIF. Blood samples were collected to assess IIF. Comparing the DIF results against the histopathology test, there was an agreement of 75% (9/12) with a Kappa index of 0.77 (P<0.001). Considering IIF, the agreement was 100% (14/14), with a Kappa index of 1.0 (P<0.001). We conclude that DIF and IIF are highly effective and were useful and effective complementary examination tests for an improvement in the diagnosis of canine PF.(AU)

O pênfigo foliáceo (PF) é considerado uma das doenças tegumentares autoimunes mais frequentes em cães. Clinicamente, caracteriza-se pela presença de pústulas, erosões e crostas. O exame histopatológico é considerado o teste diagnóstico de eleição, porém pode se mostrar inconclusivo, sobretudo quando os achados característicos da doença não são observados. Novas ferramentas diagnósticas têm sido desenvolvidas e os testes de imunofluorecência são uma valiosa alternativa. Este estudo teve como objetivo avaliar a aplicabilidade das reações de imunofluorescência direta (IFD) e indireta (IFI) para o diagnóstico do PF canino. Vinte e oito cães foram divididos em dois grupos: grupo I com 14 cães com PF e grupo II (controle) com 14 cães com piodermite superficial (um dos principais diagnósticos diferenciais do PF). Todos os animais foram submetidos à biópsia cutânea, seguida de exame histopatológico e IFD. Amostras de sangue foram coletadas para realização da IFI. Comparando-se os valores de IFD com o histopatológico, obtiveram-se valores de concordância de 75% (9/12), com índice Kappa de 0,77 (P<0,001). Já na IFI, a concordância foi de 100% (14/14), com índice Kappa de 1,0 (P<0,001). Concluiu-se, então, que a IFD e a IFI apresentaram excelentes resultados e podem ser consideradas novas alternativas diagnósticas do PF canino.(AU)

Clinical value of serum (1-3 )-β-d-glucan detection on diagnosis of deep fungal infection / 国际检验医学杂志

Objective To investigate the clinical value of serum (1-3)-β-D-glucan(BG) detection (G test) in early diagnosis of deep fungal infection .Methods 132 patients with suspected deep fungal infection in the Chongqing Emergency Medical Center from October 2015 to April 2016 were selected as the research subjects .Among them ,38 cases definitely diagnosed and suspected diagnosed deep fungal infection served as the positive group and other 94 cases were taken as the negative group .Serum BG level was measured by Jinshanchuan MB-80 microbial dynamic detection system ,and the results were compared with the fungal culture results by the body fluid fungal culture .Results The serum BG level of the positive group was (150 .8 ± 133 .2)pg/mL ,and which of the negative group was (25 .7 ± 20 .1)pg/mL ,the difference was statistically significant (t=5 .76 ,P＜0 ,05) .The sensitivity ,specificity ,positive predictive value and negative predictive value of G test were 78 .9% ,85 .1% ,68 .1% and 91 .0% respectively ;the sensitivity ,specificity ,positive predictive value and negative predictive value of fungal culture were 57 .9% ,80 .9% ,52 .3% and 82 .6%respectively .Conclusion The serum BG detection is fast ,accurate ,simple and convenient ,and has clinical application value in the early diagnosis of deep fungal infection and clinical rational medication guidance .

Overexpression of IFD6 in Candida albicans promotes biofilm formation / 第二军医大学学报

Objective: To investigate the effect of IFD6 on biofilm-forming ability of C. albicans. Methods: We constructed an IFD6-overexpressing plasmid and inserted IFD6 intact open reading frame (ORF) under the control of the MET3 promoter in pCaEXP plasmid, which was then used to transform C. albicans CAI4 by lithium acetate method. PCR was used to investigate the in situ integration of IFD6 gene and real-time PCR was used to select strains highly expressing IFD6 gene. XTT assay and confocal scanning laser microscopy were used to investigate the changes of biofilm-forming ability before and after IFD6 overexpression. We also investigated the changes of cell surface hydrophobicity (CSH) using relative CSH assay. Results: We successfully constructed IFD6-overexpressing plasmid as confirmed by restriction enzyme digestion and sequencing; IFD6-overexpressing strain was successfully established as confirmed by PCR. Real-time PCR successfully selected a strain highly expressing IFD6 gene. The results of XTT assay and confocal scanning laser microscopy both showed that IFD6-overexpressing strain had an enhanced biofilm forming ability and an increased cell surface hydrophobicity. Conclusion: Overexpression of IFD6 can enhance the biofilm forming ability of C. albicans by increasing CSH.

Diagnóstico de pneumocystis jirovecii por la técnica de reacción en cadena de la polimerasa / Pneumocystis jirovecii diagnosis by polymerase chain reaction technique

La neumonía por Pneumocystis jirovecii (PCP) es una infección respiratoria grave, considerada como una de las complicaciones más comunes en pacientes inmunocomprometidos. En Venezuela existen pocas investigaciones sobre la PCP, todas ellas realizadas usando la técnica de inmunofluorescencia directa (IFD). Actualmente es necesario disponer de métodos adicionales de detección para esta enfermedad, que en conjunto con los métodos convencionales, incrementen los rangos de sensibilidad y especificidad, para proporcionar un diagnóstico precoz. En este estudio se pretende implementar la técnica de Reacción en Cadena de la Polimerasa (PCR) para el diagnóstico de Pneumocystis jirovecii. Para ello se utilizaron 62 muestras del tracto respiratorio provenientes de pacientes con SIDA, cáncer e infección respiratoria baja sin SIDA ni cáncer. Las muestras fueron procesadas por la técnica de IFD y por la técnica de PCR anidado, usando cebadores externos (pAZ102-E y pAZ102-H) e internos (pAZ102-X y pAZ102-Y), dirigidos a la región de la subunidad mayor mitocondrial (mtLSUrRNA) del genoma de P. jirovecii, propuestos por Wakefield et al. Los resultados obtenidos por PCR fueron comparados con los de IFD (como técnica de referencia), mediante la prueba de X 2 y se calcularon valores de sensibilidad (S), especificidad (E), valores predictivos (VP), razones de verosimilitud (RV), errores y concordancia para la técnica de PCR. Se detectó P. jirovecii en 14 pacientes por la técnica de IFD y en 24 por PCR. La técnica de PCR tiene valores de S=100%; E=79,2%; VPP=58,3%; VPN=100%; RVP=4,8; RVN=0,3 y una concordancia de 84%. En conclusión, la PCR es una prueba de alto valor diagnóstico ya que predice con éxito la ausencia de la PCP cuando el resultado es negativo. Un resultado positivo no discrimina colonización de infección, por lo tanto, debe ser interpretado con precaución tomando en cuenta la clínica del paciente.

The Pneumocystis jirovecii pneumonia (PCP) is a severe respiratory infection, considered one of the most common complications in immunocompromised patients. In Venezuela there are few researches on PCP, all of them by us ing direct immunofluores cence technique (DIF). Currently, it is necessary to have ot her methods to detect this infection additionally to the use of conventional methods, in order to provide an early diagnosis with the increas e of the sensibility and specificity ranges of detection. This study aims to implement the polymerase chain reaction (PCR) for the diagnosis of Pneumocystis jirovecii Sixty two (62) clinical samples coming from AIDS, cancer and non- AIDS-non cancer low respiratory tract infections patients were processed by DIF and nested PCR techniques. The external (pAZ102-E and pAZ102-H) and internal (pAZ102-X and pAZ102-Y) primers targeting the mitochondrial Large Subunit RNA region (mtLSUrRNA) of P. jirovecii genome, proposed by Wakefield et al, were used in the nested PCR. The PCR results were compared with those of DIF (as the reference technique) using the X 2 test. Also, values of sensibility (S), specificity (E), xii predictive values (PV), verisimilitude reasons (VR), errors and agreement for The PCR techniques were calculated. P. jirovecii was detected by DIF in 14 patients and by PCR in 24 patients. PCR had values of S=100%; E=79,2%; PPV=58,3%; NPV=100%; PVR=4,8; NVR= 0 ,3 and an agreement of 84%. In conclusion, PCR is a high diagnostic value technique that successfully predicts the absence of PCP with a negative result. A positive result does not discriminate against infection colonization, therefore, it should be interpreted with caution taking into account the patient's clinic.

The effect analysis of several common used antifungal agents in invasive fungal infection with hematological malignancies / 中国实用内科杂志

Objective To explore the total treatment effect,layered diagnostic treatment effect,the relationship between effect and site of infection and common side effects of commonly used antifungal agents such as itraconazole,Voriconazole,caspofungin and amphotericin B liposome in invasive fungal infection with hematologic malignancies.Methods The clinical data of 117 cases of patients with hematologic malignancies combining with invasive fungal infection hospitalized in our department from Jan,2005 to Aug,2008 were retrospectively analyzed.Results The total effect rates of itraconazole,Voriconazole,caspofungin and amphotericin B liposome were 69.0%(40/58),77.4%(24/31),64.7%(11/17)and 63.6%(7/11)respectivley(P=0.726).In patients with pulmonary infection,the effect rates of the agents were 63.0%(17/27),85.7%(12/14),50.0%(4/8)and 62.5%(5/8)respectively(P=0.283),which itraconazole group was higher than that of the other agents.The effect rates of the 4 groups have similar rates in patients with liver and spleen candidiasis,fungemia and infections with unknown origin.The rates of 6 weeks survival were 86.2%,87.1%,70.6% and 72.7% respectively.The common side effects of itraconazole and Voriconazole were mainly gastrointestinal reaction and mild hypokalemia.There were few patients showed gastrointestinal reaction(12.1%) and hypokalemia(20.7%)in the former.Some individual showed visual abnormity(9.7%)and external vertebral body symptoms(6.4%).caspofungin showed mild toxic and side effects,which was only gastrointestinal reaction(15.4%),while they were common in amphotericin B liposome group,which were Chill and fever(81.8%),hypokalemia(100%),gastrointestinal reaction(18.2%)and liver damage(9.1%).Conclusion The total effect,layered diagnostic effect and 6 weeks survival rates were similar in itraconazole,Voriconazole,caspofungin and amphotericin B liposome in patients with hematologic malignancies combining with invasive fungal infection.