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在临床样本中使用可活性PCR检测活的Chlamydia trachomatis

Lucia Vojtech¹, Shahrokh Paktinat¹, Tiffany Luu¹

¹Department of Obstetrics and Gynecology, University of Washington, Seattle, WA, United States.

Frontiers in reproductive health

|August 21, 2023

概括

活力PCR (vPCR) 准确地区分了活的克拉米迪亚虫 (CT) 和死亡的细菌,改善了NAATs. 这种方法与培养结果有很强的相关性,可以帮助减少对CT感染不必要的抗生素使用.

科学领域:

微生物学微生物学
分子生物学分子生物学
传染性疾病传染性疾病

背景情况:

目前的核酸放大试验 (NAAT) 对于Chlamydia trachomatis (CT) 缺乏区分活跃感染和残留非活性的细菌DNA的能力.
这种限制可能导致误解结果,原因是由于过去感染或交叉污染导致的不可活性的CTDNA.
开发评估CT可行性的方法对于准确的诊断和适当的临床管理至关重要.

研究的目的:

评估可活性PCR (vPCR) 作为一种在临床样本中区分可活性CT和非可活性CT的方法的有效性.
将vPCR结果与传统培养方法和总DNA检测进行比较.

主要方法:

活性PCR (vPCR) 采用氧化染料 (PMAxx) 来排除非活性CT的DNA,以向CT ompA基因.
数字PCR被用于绝对基因组拷贝数量化.
该vPCR方法与实验室CT库存进行了验证,随后在18个阴道和25个直肠临床样本上进行了测试,这些样本先前通过NAAT呈阳性.

主要成果:

vPCR结果与已知的生存能力的实验室CT库存的预期结果密切相似.
在阴道样本中,vPCR与总DNA呈现相关性,但总DNA数量明显高于可行的CTDNA (2.2-52.6倍).
直肠样本分析表明,许多NAAT阳性样本通过总DNAPCR和vPCR呈阴性,表明没有活细菌;vPCR在一些NAAT阳性,培养阴性样本中确定了可行的CT.

结论:

vPCR提供了一种快速而简单的方法来评估临床样本中的细菌生存能力,与培养结果的相关性比总DNAPCR更强.
总DNA和vPCR之间的比率的变化凸显了临床样本中不可活性的细菌存在的不一致性.
从直肠样本的发现表明,NAAT阳性结果的很大一部分可能不表明活跃的感染,可能导致过度使用抗生素.

关键词:

克拉米迪亚病毒是什么?提取DNA 提取DNA的方法在这里,我们可以看到NAAT NAAT NAAT.这是一个PCRPCR.核酸是核酸中的一种.性传播感染性传播感染性感染性感染性感染性感染性感染性感染性感染性感染性感染性感染性感染性感染性感染性感染性感染性感染性

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