Diagnosis of latent pneumocystis jiroveci in sputum samples of patients under chemotherapy via Nested PCR
Using of chemotherapy against patients who suffer from malignancies make them susceptible to opportunistic infections. So, early diagnosis of infection and its causative factor lead to proper treatment of these patients. Pneumocystis jiroveci is one of the infections among patients who are under chemotherapy. The aim of this study is application of Nested PCR method for prompt diagnosis of pneumocystis jiroveci presence in the pulmonary samples of these patients.
Sputum samples collected from 46 patients of (KOSAR chemotherapy clinic) who suffer from cancer and were under chemotherapy treatment. All samples homogenized with 10mM and their DNA extracted. The presence of special gene of pneumocystis jiroveci (mtLSUrRNA) was evaluated with using of Nested PCR method in two phases, first phase extracted DNA out of sputum samples, and external primers Paz_102E and Paz_102H and then using first round PCR product in the preliminary phase and internal primers of Paz_102E and Paz_L2.
Among positive control samples of PCR product in a group from 346 bp and 120 bp appeared, and result of PCR of patient sputum samples showed special genus of pneumocystis jiroveci exist in the 58.69% of patient’s sputum who were under chemotherapy.
Chemotherapy suppresses the humeral and cellular immune system and makes the patients susceptible to many kinds of opportunistic infections. Pneumocystis jiroveci has a notable prevalence in pulmonary of patients who have a background of chemotherapy and Nested PCR method was recommended to early diagnosis of infections caused by this fungi.