Pneumocystis jirovecii (PCP) remains a significant cause of mortality and morbidity in patients with respiratory infections. Accurate diagnosis of PCP is still a diagnostic challenge. Hence, the main objectives were to study the incidence of Pneumocystis Jirovecii pneumonia infection among respiratory problems patients and to compare the real-time quantitative PCR technique with various diagnostic methodologies. Patients who have respiratory symptoms of PCP like breathlessness, cough, and fever were enrolled. Bronchoalveolar lavage (BAL) samples were collected and homogenized, and then smears were prepared for examination by Gomorimethanamine silver staining (GMSS), Immunofluorescent staining (IFAT), Toludine blue O (TBO), and Giemsa staining. Further, RT-PCR was also performed for the detection of PCP. The mean patients’ age was 52 (SD ± 16) years. 41% were female, and 59% of the patients were male. Weight loss (80%), fever (92%), cough (100%), and dyspnea (76%) were the most common complaints. Twenty-eight patients have been diagnosed with pulmonary infiltrates using chest X-ray. Out of 100 patients, 35% were positive for PCP. The organism was detected using IFAT in all the 35 specimens, 15 of 35 (42.86%) by GMSS, 8 of 35 (17.6%) by Giemsa stain, and 1 of 35 (2.8%) was detected by TBO stains. RT-PCR showed that 39 patients was found to be positive for PCP. Thirty-five of these 39 patients had a positive IFAT (89.74%); the IFAT was negative or undefined in 4 samples. All 39 patients (100%) had signs and symptoms for PCP. Our results suggest that RT-PCR is still the most highly sensitive method for Pneumocystis Jirovecii detection. In poor resource settings where RT-PCR and IFAT is not available, diagnosis of Pneumocystis jirovecii pneumonia remains a complicated issue. In settings where RT-PCR & IFAT are not available, GMSS staining may be the next best choice to detect PCP.

继发性肺囊虫（PCP）仍然是呼吸道感染患者死亡和发病的重要原因。准确诊断PCP仍然是诊断难题。因此，主要目的是研究吉氏肺孢子虫的发生率呼吸系统疾病患者的肺炎感染，并将实时定量PCR技术与各种诊断方法进行比较。招募有PCP呼吸道症状如呼吸困难，咳嗽和发烧的患者。收集支气管肺泡灌洗液（BAL）并匀浆，然后涂片涂片，以进行戈莫胺丹银染色（GMSS），免疫荧光染色（IFAT），甲苯胺蓝O（TBO）和Giemsa染色检查。此外，还进行了RT-PCR以检测PCP。患者的平均年龄为52（SD±16）岁。女性占41％，男性占59％。体重减轻（80％），发烧（92％），咳嗽（100％）和呼吸困难（76％）是最常见的主诉。使用胸部X光检查已诊断出28例肺部浸润。在100名患者中，PCP阳性率为35％。在所有35个样本中使用IFAT检测到该生物，通过GMSS检测到35个样本中的15个（占42.86％），通过Giemsa染色检测35个样本中的8个（占17.6％），通过TBO染色检测到35个样本中的1个（2.8％）。逆转录-聚合酶链反应显示39例PCP阳性。这39例患者中有35例IFAT阳性（89.74％）；IFAT在4个样本中为负或不确定。所有39例患者（100％）都有PCP的体征和症状。我们的结果表明，RT-PCR仍然是最灵敏的方法 IFAT在4个样本中为负或不确定。所有39例患者（100％）都有PCP的体征和症状。我们的结果表明，RT-PCR仍然是最灵敏的方法 IFAT在4个样本中为负或不确定。所有39例患者（100％）都有PCP的体征和症状。我们的结果表明，RT-PCR仍然是最灵敏的方法肺孢菌的罗非鱼的检测。在无法获得RT-PCR和IFAT的资源匮乏的地区，吉氏肺孢子虫肺炎的诊断仍然是一个复杂的问题。在无法使用RT-PCR和IFAT的环境中，GMSS染色可能是检测PCP的次佳选择。