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首页> 外文期刊>Laboratory Animal Science >Detection of Pneumocystis carinii in rats by polymerase chain reaction: comparison of lung tissue and bronchoalveolar lavage specimens.
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Detection of Pneumocystis carinii in rats by polymerase chain reaction: comparison of lung tissue and bronchoalveolar lavage specimens.

机译：聚合酶链反应检测大鼠卡氏肺孢子虫：肺组织和支气管肺泡灌洗标本的比较。

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摘要

The efficiency of detection of P. carinii in rats by polymerase chain reaction (PCR) of DNA extracted from 2 sampling locations, lung tissue and bronchoalveolar lavage, was determined. Naturally infected F344 rats were allotted to groups to intercollate the investigation of several variables, including non-immunosuppressed rats, rats subjected to a timed induction sequence of 1-4 weeks of immunosuppression, and 2 immunosuppressants: a corticosteroid and cyclophosphamide. PCR amplified a 357-base pair region contained within the gene encoding the large ribosomal RNA subunit of P. carinii mitochondrial DNA. The identity of the PCR product was confirmed by Southern blot analysis with an oligonucleotide probe. In a comparison of lung bronchoalveolar lavage specimens after immunosuppression, P. carinii was detected by PCR in 100% of lung tissue but in only 87.5% of the lavage specimens. Lung tissue of 3 animals was test-positive when the corresponding lavage specimen was negative by PCR analysis. PCRdetected P. carinii in both types of specimens from the same 2 of 3 non-immunosuppressed rats. Overall there was 88% agreement of PCR results between the 2 sampling techniques. The difference in diagnostic outcome for the 2 specimen types was not statistically significant. It is concluded that both specimen types were adequate for PCR detection of P. carinii in rats.
机译：确定了通过聚合酶链反应（PCR）检测从两个采样位置，肺组织和支气管肺泡灌洗液中提取的DNA的大鼠卡氏疟原虫的效率。将自然感染的F344大鼠分配到各组，以相互协作研究多个变量，包括非免疫抑制的大鼠，进行1-4周免疫抑制的定时诱导序列的大鼠和2种免疫抑制剂：皮质类固醇和环磷酰胺。 PCR扩增了编码卡氏疟原虫线粒体DNA大核糖体RNA亚基的基因中包含的357个碱基对区域。通过使用寡核苷酸探针的Southern印迹分析来确认PCR产物的身份。在对免疫抑制后的肺支气管肺泡灌洗标本进行比较时，通过PCR在100％的肺组织中检出了卡氏疟原虫，但在87.5％的灌洗标本中。通过PCR分析，当相应的灌洗标本为阴性时，3只动物的肺组织呈阳性反应。在来自3只非免疫抑制大鼠的相同2只中，两种类型的标本均通过PCR检测到卡氏疟原虫。总体而言，两种采样技术之间的PCR结果一致性为88％。两种样本类型的诊断结果差异无统计学意义。结论是，两种标本类型都足以用于大鼠中卡氏疟原虫的PCR检测。

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