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首页> 外文期刊>Clinical and diagnostic laboratory immunology >Diversity of host species and strains of Pneumocystis carinii is based on rRNA sequences.
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Diversity of host species and strains of Pneumocystis carinii is based on rRNA sequences.

机译：卡氏肺孢子虫宿主物种和菌株的多样性基于rRNA序列。

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We have amplified by PCR Pneumocystis carinii cytoplasmic small-subunit rRNA (variously referred to as 16S-like or 18S-like rRNA) genes from DNA extracted from bronchoalveolar lavage and induced sputum specimens from patients positive for P. carinii and from infected ferret lung tissue. The amplification products were cloned into pUC18, and individual clones were sequenced. Comparison of the determined sequences with each other and with published rat and partial human P.carinii small-subunit rRNA gene sequences reveals that, although all P. carinii small-subunit rRNAs are closely related (approximately 96% identity), small-subunit rRNA genes isolated from different host species (human, rat, and ferret) exhibit distinctive patterns of sequence variation. Two types of sequences were isolated from the infected ferret lung tissue, one as a predominant species and the other as a minor species. There was 96% identity between the two types. In situ hybridization of the infected ferret lung tissue with oligonucleotide probes specific for each type revealed that there were two distinct strains of P. carinii present in the ferret lung tissue. Unlike the ferret P. carinii isolates, the small-subunit rRNA gene sequences from different human P. carinii isolates have greater than 99% identity and are distinct from all rat and ferret sequences so far inspected or reported in the literature. Southern blot hybridization analysis of PCR amplification products from several additional bronchoalveolar lavage or induced sputum specimens from P. carinii-infected patients, using a 32P-labeled oligonucleotide probe specific for human P. carinii, also suggests that all of the human P. carinii isolates are identical. These findings indicate that human P. carinii isolates may represent a distinct species of P. carinii distinguishable from rat and ferret P. carinii on the basis of characterization of small-subunit rRNA gene sequences.
机译：我们已经通过PCR扩增了卡氏肺孢子虫胞质小亚基rRNA（也称为16S样或18S样rRNA）基因，该基因是从支气管肺泡灌洗液提取的DNA中提取的，并从卡氏疟原虫阳性患者和受感染的雪貂肺组织诱导的痰标本中提取。将扩增产物克隆到pUC18中，并对单个克隆进行测序。将确定的序列彼此之间以及与已发表的大鼠和部分人卡氏毕赤酵母小亚基rRNA基因序列进行比较，结果表明，尽管所有卡氏毕赤酵母小亚基rRNA都密切相关（大约96％相同），但是小亚基rRNA从不同宿主物种（人类，大鼠和雪貂）分离的基因显示出独特的序列变异模式。从受感染的雪貂肺组织中分离出两种类型的序列，一种为优势种，另一种为次要种。两种类型之间有96％的同一性。用对每种类型特异的寡核苷酸探针对受感染的雪貂肺组织进行原位杂交表明，雪貂肺组织中存在两种不同的卡氏疟原虫菌株。与雪貂分离株不同，来自不同人类卡氏分离株的小亚基rRNA基因序列具有超过99％的同一性，并且与迄今为止在文献中检查或报道的所有大鼠和雪貂序列不同。使用对人卡氏疟原虫特异的32P标记寡核苷酸探针，对来自卡氏疟原虫感染患者的数个其他支气管肺泡灌洗液或诱导的痰标本进行PCR扩增产物的Southern印迹杂交分析也表明，所有人卡氏疟原虫分离株完全一样这些发现表明，基于小亚基rRNA基因序列的表征，人类卡氏毕赤酵母分离物可以代表与大鼠和雪貂卡氏毕赤酵母不同的卡氏疟原虫物种。

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