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利用聚合酶链式反应(PCR)和显微镜检查法检测免疫抑制大鼠鼻拭子中的肺孢子菌。

Detection of Pneumocystis in the nasal swabs of immune-suppressed rats by use of PCR and microscopy.

作者信息

Can Hüseyin, Caner Ayşe, Döşkaya Mert, Değirmenci Aysu, Karaçalı Sabire, Polat Ceylan, Gürüz Yüksel, Uner Ahmet

机构信息

Ege University Medical School, Department of Parasitology, Bornova/Izmir, Turkey.

出版信息

Med Sci Monit Basic Res. 2013 Feb 13;19:62-7. doi: 10.12659/MSMBR.883777.

Abstract

BACKGROUND

Detection of Pneumocystis jiroveci colonization in lungs or oral samples due to high sensitivity of PCR methods results in undue treatment of patients without any symptoms of Pneumocystis pneumonia. The aim of the present study is to demonstrate Pneumocystis carinii in rats, immune suppressed by oral and subcutaneous administration of dexamethasone.

MATERIAL/METHODS: Blood, oral, nasal and eye swabs were collected prior to immune suppression and 2, 6, 12 weeks after administration of dexamethasone. Also, samples were collected from lung, heart, liver, kidney, diaphragm, brain, spleen, tongue, muscle, eye, intestine, and feces. Cysts and trophozoites were investigated in stained slides and MSG gene was detected by PCR.

RESULTS

The results showed that weight loss is significantly higher in rats administered oral dexamethasone (P<0.05). Microscopy was positive only in lungs of rats orally administered dexamethasone. PCR was positive in lungs and oral swabs of rats prior to the administration of dexamethasone. After the administration of dexamethasone, the MSG gene was detected in oral swabs, lungs, spleen, kidney and (for the first time) in nasal swabs. PCR was positive in nasal swabs during the second and sixth weeks of oral and subcutaneous administration of dexamethasone, respectively.

CONCLUSIONS

Presence of P. jiroveci in nasopharyngeal aspirate, oropharyngeal wash, oral swab, induced sputum or BAL, and absence in nasal swab in a patient without symptoms of PCP may support clinician's decision regarding colonization. Overall, detection of P. carinii in nasal swabs of rats by PCR demonstrated that nasal sampling can be used for the diagnosis of Pneumocystis pneumonia.

摘要

背景

由于聚合酶链反应(PCR)方法具有高灵敏度,在肺部或口腔样本中检测到耶氏肺孢子菌定植,导致对没有任何肺孢子菌肺炎症状的患者进行过度治疗。本研究的目的是在经口服和皮下给予地塞米松免疫抑制的大鼠中证实卡氏肺孢子菌的存在。

材料/方法:在免疫抑制前以及给予地塞米松后2周、6周和12周采集血液、口腔、鼻腔和眼部拭子。此外,还从肺、心脏、肝脏、肾脏、膈肌、大脑、脾脏、舌头、肌肉、眼睛、肠道和粪便中采集样本。在染色载玻片上研究包囊和滋养体,并通过PCR检测MSG基因。

结果

结果显示,口服地塞米松的大鼠体重减轻明显更高(P<0.05)。显微镜检查仅在口服地塞米松的大鼠肺部呈阳性。PCR在给予地塞米松前大鼠的肺部和口腔拭子中呈阳性。给予地塞米松后,在口腔拭子、肺部、脾脏、肾脏以及(首次)鼻腔拭子中检测到MSG基因。在口服和皮下给予地塞米松的第二周和第六周,鼻腔拭子中的PCR分别呈阳性。

结论

在没有肺孢子菌肺炎症状的患者中,鼻咽抽吸物、口咽冲洗液口腔拭子、诱导痰或支气管肺泡灌洗中有耶氏肺孢子菌,而鼻腔拭子中没有,这可能支持临床医生关于定植的判断。总体而言,通过PCR在大鼠鼻腔拭子中检测到卡氏肺孢子菌表明,鼻腔采样可用于肺孢子菌肺炎的诊断。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8555/3638709/0d1a48de1865/medscimonitbasicres-19-62-g001.jpg

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