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首页> 外文期刊>Clinical and diagnostic laboratory immunology >Analysis of Chlamydia pneumoniae Growth in Cells by Reverse Transcription-PCR Targeted to Bacterial Gene Transcripts
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Analysis of Chlamydia pneumoniae Growth in Cells by Reverse Transcription-PCR Targeted to Bacterial Gene Transcripts

机译：靶向细菌基因转录本的逆转录PCR技术分析细胞中肺炎衣原体的生长

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Chlamydia pneumoniae is an obligate intracellular bacterium and has a unique development cycle consisting of an elementary body (EB) and reticular body (RB). EBs survive in extracellular environments as well as infect susceptible host cells. However, EBs display no measurable metabolic activity. In contrast, RBs are metabolically active and can replicate in a host cell but are noninfectious. Therefore, analysis of C. pneumoniae growth in infected cells by conventional bacterial culture may not permit sufficient information about growth of the bacteria in cells. In this study, therefore, we examined the usefulness of the reverse transcription (RT)-PCR method for analysis of bacterial transcripts to evaluate C. pneumoniae growth in HEp-2 cells because the levels of bacterial gene transcripts are known to show the metabolic activity of bacteria. The transcripts for the C. pneumoniae hsp60 gene and 16S rRNA in the cells were easily detected just after infection, followed by a marked increase. In contrast, pyk and omcB transcripts slowly increased after a latent period. The hydrocortisone treatment of C. pneumoniae-infected cells induced an increase of all bacterial transcripts tested compared with the control group. The treatment of the infected cells with the antibiotic minocycline showed a selective inhibition of bacterial gene transcripts, even though the complete inhibition of EB production determined by the bacterial culture assay was evident. These results indicate that the determination of bacterial gene transcripts by RT-PCR might be a powerful method to analyze in detail growth of C. pneumoniae in host cells, particularly altered bacterial growth caused by agents such as antimicrobials.
机译：肺炎衣原体是专性的细胞内细菌，具有独特的发育周期，由基本体（EB）和网状体（RB）组成。 EB在细胞外环境以及感染易感宿主细胞中生存。但是，EBs没有可测量的代谢活性。相反，RB具有代谢活性，可以在宿主细胞中复制，但无感染性。因此，分析 C。常规细菌培养在感染细胞中肺炎的生长可能无法提供有关细菌在细胞中生长的足够信息。因此，在这项研究中，我们研究了逆转录（RT）-PCR方法对细菌转录本进行分析以评估C的有效性。由于已知细菌基因转录物的水平显示出细菌的代谢活性，因此HEp-2细胞中出现了肺炎。 C的成绩单。感染后很容易检测到肺炎hsp60 基因和16S rRNA，然后明显升高。相反， pyk 和 omcB 转录本在潜伏期后缓慢增加。氢化可的松治疗 C。与对照组相比，肺炎链球菌感染的细胞诱导了所有细菌转录本的增加。即使用细菌培养分析法确定了对EB产生的完全抑制作用，用抗生素米诺环素处理感染的细胞也显示出对细菌基因转录的选择性抑制。这些结果表明，通过RT-PCR确定细菌基因转录本可能是一种详细分析 C的强大方法。宿主细胞中的肺炎，尤其是由诸如抗菌剂之类的药物引起的细菌生长改变。

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