Estudo de novos marcadores serológicos para o diagnóstico da pneumonia por pneumocystis jirovecii
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2012
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Pneumocystis jirovecii (P.j.) é um organismo oportunista responsável por uma das infecções mais severas em doentes imunocomprometidos, a pneumonia por Pneumocystis ou pneumocistose (PPc). O diagnóstico laboratorial é baseado em colorações histoquímicas, imunofluorescência indirecta com anticorpos monoclonais e técnica de PCR em secreções pulmonares. Estudos sugerem que Krebs von den Lungen-6 (KL-6), e S-adenosilmetionina (SAM), poderão ser utilizados como indicadores sensíveis para o diagnóstico da PPc. O objectivo do presente trabalho é a avaliação destes dois marcadores no diagnóstico serológico da PPc, utilizando dois ensaios ELISA. Soro e espécimes pulmonares foram obtidos de 145 doentes (1997-2012). O soro de 37 dadores foi utilizado como controlo negativo.
Dos 145 doentes, 92 eram casos confirmados de PPc (mediana: KL-6 969 U/mL; SAM 38 nmol/L) e 53 possuíam outra patologia pulmonar (23 colonizados por P.j. e 30 não infectados por P.j.) (mediana: KL-6 437 U/mL; SAM 60 nmol/L). A mediana determinada para os doentes colonizados foi de 409 U/mL (KL-6) e 55 nmol/L (SAM) e, nos doentes sem infecção com P.j. foi de 440 U/mL (KL-6) e 60 nmol/L (SAM). Nos dadores de sangue, a mediana de KL-6 e SAM foi, respectivamente, de 230 U/mL e 75 nmol/L. Níveis altos de KL-6, no soro, (> 590 U/mL) foram estatisticamente associados aos casos de PPc (76%, P < 0,001). Estes resultados sugerem que o KL-6 poderá ser um candidato para o diagnóstico serológico de PPc, principalmente quando utilizado em associação com outros marcadores.
Pneumocystis jirovecii (P.j.) is an opportunistic pathogen responsible for one of the most severe infections in immunocompromised patients, Pneumocystis pneumonia (PcP). The laboratory diagnosis of PcP is based on cytochemical staining, indirect immunofluorescence with monoclonal antibodies and PCR on pulmonary secretions. Studies suggest that Krebs von den Lungen-6 (KL-6) and S-adenosyl Methionine (SAM) may be used as sensitive indicators for PcP diagnosis. The aim of the present work is to evaluate these two potential serological markers, using two different ELISA assays. Sera and pulmonary specimens were obtained from 145 Portuguese patients, for diagnostic purpose (1997-2012). Sera were collected from 37 blood donors (negative control group). Of the 145 patients, 92 were PcP positive (median: KL-6 969 U/mL; SAM 38 nmol/L) and 53 were PcP negative (23 were colonized by P.j. and 30 were not infected by P.j.) (median: KL-6 437 U/mL; SAM 60 nmol/L). The median serologic markers levels detected for the colonized carriers were 409 U/mL (KL-6) and 55 nmol/L (SAM) while in the patients without P.j. infection were 440 U/mL (KL-6) and 60 nmol/L (SAM). In the blood donors the median KL-6 level was 230 U/mL, while the median SAM serological level was 75 nmol/L. Sera presenting high levels of KL-6 (>590 U/mL) were statistically associated with PcP cases (76%, P <0.001). These results suggest that KL-6 might be a candidate to use in PcP serologic diagnosis, mostly when used in association with other markers.
Pneumocystis jirovecii (P.j.) is an opportunistic pathogen responsible for one of the most severe infections in immunocompromised patients, Pneumocystis pneumonia (PcP). The laboratory diagnosis of PcP is based on cytochemical staining, indirect immunofluorescence with monoclonal antibodies and PCR on pulmonary secretions. Studies suggest that Krebs von den Lungen-6 (KL-6) and S-adenosyl Methionine (SAM) may be used as sensitive indicators for PcP diagnosis. The aim of the present work is to evaluate these two potential serological markers, using two different ELISA assays. Sera and pulmonary specimens were obtained from 145 Portuguese patients, for diagnostic purpose (1997-2012). Sera were collected from 37 blood donors (negative control group). Of the 145 patients, 92 were PcP positive (median: KL-6 969 U/mL; SAM 38 nmol/L) and 53 were PcP negative (23 were colonized by P.j. and 30 were not infected by P.j.) (median: KL-6 437 U/mL; SAM 60 nmol/L). The median serologic markers levels detected for the colonized carriers were 409 U/mL (KL-6) and 55 nmol/L (SAM) while in the patients without P.j. infection were 440 U/mL (KL-6) and 60 nmol/L (SAM). In the blood donors the median KL-6 level was 230 U/mL, while the median SAM serological level was 75 nmol/L. Sera presenting high levels of KL-6 (>590 U/mL) were statistically associated with PcP cases (76%, P <0.001). These results suggest that KL-6 might be a candidate to use in PcP serologic diagnosis, mostly when used in association with other markers.
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Orientação : Francisco Vaz de Carvalho Esteves ; coorientação : Olga Maria Guerreiro de Matos
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ENGENHARIA BIOTECNOLÓGICA, BIOTECHNOLOGY ENGINEERING, DOENÇAS INFECCIOSAS, INFECTIOUS DISEASES, PNEUMOLOGIA, PNEUMOLOGY, MESTRADO EM ENGENHARIA BIOTECNOLÓGICA