Blood and sputum specimens were collected from 98 patients before they received the corticoid or antibiotics treatment. Informed consent was obtained from all patients.
Sputum specimens were obtained from patient by spontaneous production. In order to avoid contamination of oral microbe, patients first gargled saline three times, prior to coughing up the sputum from deep respiratory tract when collecting specimens. Sputum specimens were promptly sent to the central laboratory of the hospital for bacterial culture and quantization. Co-infecting bacteria was analyzed using the automated bacterial identification system (ATB system, BioMérieux, Marcy-l’Étoile, France) following the bacterial or fungi culture. P. jirovecii organisms were identified microscopically by a modified Giemsa staining (Diff-Quik) method and a modified Gomori’s methenamine silver nitrate staining method as previously described.13 ,14
The blood samples were used for CD4+ T-cell measurement.12 (link) Sera were analyzed on BD FACSCalibur and the test kit. The normal critical value of CD4+ T-cell in blood was set to 410 cells/mm3 according to the reference of test kit. Patients will be considered as immunocompetent when CD4 cell >410 cells/mm3, and they will be identified as immunocompromised when the CD4+ T-cell counts <410 cells/mm3.
Sputum specimens were obtained from patient by spontaneous production. In order to avoid contamination of oral microbe, patients first gargled saline three times, prior to coughing up the sputum from deep respiratory tract when collecting specimens. Sputum specimens were promptly sent to the central laboratory of the hospital for bacterial culture and quantization. Co-infecting bacteria was analyzed using the automated bacterial identification system (ATB system, BioMérieux, Marcy-l’Étoile, France) following the bacterial or fungi culture. P. jirovecii organisms were identified microscopically by a modified Giemsa staining (Diff-Quik) method and a modified Gomori’s methenamine silver nitrate staining method as previously described.13 ,14
The blood samples were used for CD4+ T-cell measurement.12 (link) Sera were analyzed on BD FACSCalibur and the test kit. The normal critical value of CD4+ T-cell in blood was set to 410 cells/mm3 according to the reference of test kit. Patients will be considered as immunocompetent when CD4 cell >410 cells/mm3, and they will be identified as immunocompromised when the CD4+ T-cell counts <410 cells/mm3.