Características clínicas, métodos diagnósticos y evolución de la criptococosis extrameníngea en personas viviendo con VIH / Clinical features, diagnosis methods and evolution in non meningeal cryptococcosis in people living with VIH

La criptococosis es una micosis grave que se manifiesta, en el 90% de los casos, como una meningoencefalitis, especialmente en las personas con VIH. El objetivo de este estudio es describir los casos de criptococosis extrameníngea en personas viviendo con VIH y conocer cuántas de estas padecen compromiso meníngeo concomitante. Además, determinar la relación con el título de antígeno polisacárido capsular de Cryptococcus en suero. Se realizó un estudio retrospectivo, observacional y analítico. Se incluyeron personas viviendo con VIH cuyo diagnóstico inicial de criptococosis se había realizado a partir de muestras extrameníngeas en el período comprendido entre 2012 y 2019. Los pacientes se dividieron en dos grupos. Grupo 1, pacientes sin compromiso meníngeo; Grupo 2, aquellos que finalmente tenían compromiso del SNC. De un total de 531 criptococosis registradas en ese período, se incluyeron 113 pacientes (21%), de los cuales en 58 se comprobó el compromiso meníngeo. No se observaron diferencias significativas en cuanto a la mortalidad entre ambos grupos.Ninguno de los pacientes con antigenemia por LFA (antígeno capsular en suero por inmunocromatografía) positiva, pero con antigenemia por aglutinación de partículas de látex (AL) negativa, tuvo compromiso meníngeo. Se observó que títulos de antígeno para Cryptococcus en suero por AL mayor o igual a 1/100 se correlacionaron con un aumento de 30 veces en la posibilidad de padecer meningitis. En todos los casos se debe descartar el compromiso del SNC. La AL sigue siendo una prueba útil y complementaria, debido a que en los casos con AL negativa no se observó compromiso meníngeo

Cryptococcosis is a serious mycosis that manifests itself, in 90% of cases, as meningoencephalitis, especially in AIDS patients. The objective of this study is to describe the extra-meningeal cases of cryptococcosis in people living with HIV and to know how many of them suffer from concomitant meningeal involvement. Also, to determine its relationship with the Cryptococcus capsular polysaccharide antigen titer in serum.A retrospective, observational and analytical study was carried out. HIV-positive patients whose initial diagnosis had been made from extrameningeal samples in the period between 2012 and 2019 were included. The patients were divided into 2 groups. Group 1: patients without meningeal involvement; group 2: those who finally had CNS involvement.Of a total of 531 cryptococcosis registered in this period, 113 patients (21%) were included, of whom meningeal involvement was confirmed in 58. No significant differences were observed in terms of mortality in both groups.None of the patients with positive LFA antigenemia (Capsular antigen detection by lateral Flow assay) but negative latex particle agglutination (LA) antigenemia had meningeal involvement. LFA was found to be highly sensitive and allows early diagnosis, but it does not replace other diagnostic procedures.Serum Cryptococcus antigen titers for by LA greater than or equal to 1/100 were found to correlate with a 30-fold increase in the likelihood of meningitis.In all cases, CNS involvement must be ruled out. LA continues to be a useful and complementary test, because in cases with negative LA, no meningeal involvement was observed

Estudo piloto de avaliação de um teste rápido imunocromatográfico para o diagnóstico de sífilis gestacional / Pilot evaluation of a rapid immunochromatographic test for the diagnosis of gestational syphilis

A sífilis gestacional é um problema global de saúde pública e é uma das mais comuns causas de efeitos adversos durante a gravidez devido à ausência ou inadequação do tratamento. Estabelecer um diagnóstico de sífilis durante o pré-natal, evita a transmissão de Treponema pallidum para a criança. Objetivo: o objetivo deste estudo foi avaliar o desempenho de OL Syphilis (OrangeLife, Brasil), um teste imunocromatográfico rápido, para o diagnóstico de sífilis gestacional. Métodos: Um total de 185 mulheres grávidas no pré-natal foram avaliadas por sífilis OL. Os resultados foram comparados com os métodos tradicionais: Laboratório de Pesquisa de Doenças Venéreas e Reaginas de Plasma Rápido (VDRL e RPR) como ensaios de seleção e FTA-ABS como teste confirmatório. Resultados: A prevalência de sífilis nessa população foi de 6,49% (IC 3,40 a 11,06%). A sensibilidade do teste rápido (TR) foi de 91,67% (IC95% 61,52 a 99,79%) e a especificidade foi de 100% (95%IC 97,89 a 100%). O PPV foi 100% (95%CI 71,51 a 100%) e o VPL foi de 99,43 (95%CI 96,84 a 100%). O acordo medido pelo coeficiente Kappa foi de 0,954 (IC95% 0,863 a 1,000). Conclusão: O teste OL Syphilis poderia ser usado no rastreio de mulheres grávidas, fornecendo diagnóstico rápido, aumentando a probabilidade de ter a doença diagnosticada e oportuna, evitando as consequências devastadoras da sífilis congênita.

Gestational syphilis is a global public health problem and one of the most common causes of adverse effects during pregnancy due to absence or inadequacy of treatment. Establishing a diagnosis of syphilis during prenatal care prevents the transmission of Treponema pallidum to the child. Objective: The objective of this study was to evaluate the performance of OL Syphilis (OrangeLife, Brazil), a rapid immunochromatographic test for gestational syphilis diagnosis. Methods: A total of 185 pregnant women in prenatal care were evaluated by OL Syphilis. The results were compared by traditional methods: Venereal Disease Research Laboratory and Rapid Plasma Reagin (VDRL and RPR) for screening and fluorescent treponemal antibody absorption test (FTA-Abs) for confirmation. Results: The prevalence of syphilis in this population was 6.49% (95%CI 3.40 to 11.06%). Rapid Test (RT) sensitivity was 91.67% (95%CI 61.52 to 99.79%) and specificity was 100% (95%CI 97.89 to 100%). Positive predictive value was 100% (95%CI 71.51 to 100%) and Negative predictive value was 99.43% (95%CI 96.84 to 100%). The agreement measured by Kappa coefficient was 0.954 (95%CI 0.863 to 1.000). Conclusion: The OL Syphilis test could be used for screening pregnant women, thus providing rapid diagnosis, increasing the probability of diagnosis and timely treatment, and preventing the devastating consequences of congenital syphilis.

Construction, transfection and production of recombinant vigilin in mammalian expression system

Vigilin is an abundant, highly conserved, ubiquitous protein containing 15 related, but non-identical, K-homolous nucleic acid binding domains. The construction, transfection and production of recombinant vigilin in mammalian expression system were investigated. The whole length of vigilin was amplified by polymerase chain reaction [PCR] and ligated to pCEP-PU vector. The recombinant construct pCEP-PU with vigilin was produced and transfected into Human embryonic kidney cells in a specific culture medium. The secreted recombinant vigilin was purified from the serum-free medium to homogeneity by affinity chromatography. The conditioned and purified media were tested for the presence of vigilin by sodium dodecyl sulfate polyacrylamide gel electrophoresis [SDS-PAGE] and immunoblotting with specific antibody. An immunoreactive band with an apparent molecular mass of approximately 140 kDa was detected. Immunofluorescence staining of transfected cells with vigilin demonstrated that recombinant vigilin molecules are localized in the nucleus and cytoplasm. In summary, the purified recombinant vigilin will facilitate future studies that address the structure and function of vigilin