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Laboratory diagnosis of Mycoplasma pneumoniae infection. 4. Antigen capture and PCR-gene amplification for detection of the Mycoplasma: problems of clinical correlation.

机译:肺炎支原体感染的实验室诊断。 4.用于支原体检测的抗原捕获和PCR基因扩增:临床相关性问题。

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摘要

Direct detection assays for Mycoplasma pneumoniae were established by PCR amplification of short sequences within the foot protein/adhesin (P1) gene and the 16S ribosomal RNA gene. Specificity and sensitivity was excellent, no hybridization was observed with M. genitalium and other human Mycoplasma species. In nose and throat washings from subjects with respiratory infection a pattern of high counts (c.f.u./ml) of M. pneumoniae (deduced from the amount of amplified PCR product), and a positive antigen capture assay, was found in 83% of subjects with serological evidence of current infection with M. pneumoniae. A small proportion of subjects with serological patterns suggesting infection in the more distant past had positive PCR assays. This was considered to represent either persistence of the organism from a previous infection or perhaps transient carriage during a reinfection, without substantial change in antibody response. PCR-based assay of M. pneumoniae offers a powerful, rapid, and sensitive substitute for culture of the mycoplasma. Antigen capture, while less sensitive than PCR, offers the advantage that it is more often positive with samples from current infection and requires less stringent laboratory organization to contain false positive results. We conclude however that the laboratory diagnosis of a chosen clinical episode should not rest on the PCR or Ag-EIA assays alone, but must also include antibody assays to confirm whether infection is current or represents persistence from past exposure.
机译:通过对足蛋白/粘附素(P1)基因和16S核糖体RNA基因中的短序列进行PCR扩增,建立了肺炎支原体的直接检测方法。特异性和敏感性极好,未观察到生殖器支原体和其他人类支原体物种的杂交。在患有呼吸道感染的受试者的鼻腔和咽喉冲洗物中,肺炎支原体的计数很高(cfu / ml)(从扩增的PCR产物的量推导),并且在83%的受试者中发现了阳性的抗原捕获测定当前感染肺炎支原体的血清学证据。血清学模式表明在较远的过去感染的一小部分受试者进行了阳性PCR检测。这被认为代表了先前感染的生物体的持久性,或者代表了再感染过程中的短暂携带,而抗体反应却没有实质性变化。基于PCR的肺炎支原体测定为支原体培养提供了强大,快速和灵敏的替代品。抗原捕获虽然不如PCR灵敏,但具有以下优点:对于当前感染的样本,抗原捕获更常为阳性,并且不需要严格的实验室组织即可包含假阳性结果。然而,我们得出的结论是,所选临床发作的实验室诊断不应仅依靠PCR或Ag-EIA分析,而还必须包括抗体分析以确认感染是当前的还是代表过去暴露的持续存在。

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