DNA-based culture-independent analysis detects the presence of group a streptococcus in throat samples from healthy adults in Japan.
DNA-based culture-independent analysis detects the presence of group a streptococcus in throat samples from healthy adults in Japan.
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DOI：
10.1186/s12866-016-0858-5
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发表时间：
2016-10-11
期刊：
影响因子：
4.2
通讯作者：
Nakagawa I
中科院分区：
文献类型：
--
作者：
Kulkarni T;Aikawa C;Nozawa T;Murase K;Maruyama F;Nakagawa I
A 组链球菌（GAS；化脓性链球菌）会引起人类一系列轻度至重度感染。它还可以在无症状的健康人中定殖。因此，研究健康人群中的 GAS 携带非常重要，因为它的携带可能导致随后的疾病表现、社区中的克隆传播和/或生物体的多样化。咽拭子培养是 GAS 检测的金标准方法。先进的独立于培养的方法可以直接从临床样本中快速有效地检测微生物。我们使用依赖培养和不依赖培养的方法调查了日本健康成年人咽拭子样本中 GAS 的存在情况。从 148 名健康志愿者身上采集了两份咽拭子样本。其中一个在选择性培养基上培养，而从另一个中提取的总DNA则用两对GAS特异性引物对进行聚合酶链式反应（PCR）扩增：一对是新设计的16S rRNA特异性引物对，另一对是先前描述的V-Na+-ATPase引物对。尽管通过依赖培养的方法，148 个样本中只有 5 个 (3.4%) 呈 GAS 阳性，但通过不依赖培养的方法，146 个样本 (98.6%) 的 GAS DNA 呈阳性。为了通过 emm 分型获得血清型信息，我们使用新设计的 emm 引物进行了巢式 PCR。我们在 25 个 (16.9%) 样本中检测到了四种不同的 emm 类型，这些类型与日本与 GAS 相关疾病相关的常见 emm 类型不同。在健康志愿者中检测到的不同 emm 类型表明，独特 emm 类型的存在可能与 GAS 携带有关。我们的结果表明，应考虑使用独立于培养的方法来分析健康宿主中的 GAS，以便更好地了解这些生物体。本研究中使用的 GAS 特异性引物（16S rRNA 和 V-Na+-ATPase）可用于估计人体内最大潜在的 GAS 携带量。本文的在线版本 (doi:10.1186/s12866-016-0858-5) 包含补充材料，可供授权用户使用。
Group A Streptococcus (GAS; Streptococcus pyogenes) causes a range of mild to severe infections in humans. It can also colonize healthy persons asymptomatically. Therefore, it is important to study GAS carriage in healthy populations, as carriage of it might lead to subsequent disease manifestation, clonal spread in the community, and/or diversification of the organism. Throat swab culture is the gold standard method for GAS detection. Advanced culture-independent methods provide rapid and efficient detection of microorganisms directly from clinical samples. We investigated the presence of GAS in throat swab samples from healthy adults in Japan using culture-dependent and culture-independent methods. Two throat swab samples were collected from 148 healthy volunteers. One was cultured on selective medium, while total DNA extracted from the other was polymerase chain reaction (PCR) amplified with two GAS-specific primer pairs: one was a newly designed 16S rRNA-specific primer pair, the other a previously described V-Na+-ATPase primer pair. Although only 5 (3.4 %) of the 148 samples were GAS-positive by the culture-dependent method, 146 (98.6 %) were positive for the presence of GAS DNA by the culture-independent method. To obtain serotype information by emm typing, we performed nested PCR using newly designed emm primers. We detected the four different emm types in 25 (16.9 %) samples, and these differed from the common emm types associated with GAS associated diseases in Japan. The different emm types detected in the healthy volunteers indicate that the presence of unique emm types might be associated with GAS carriage. Our results suggest that culture-independent methods should be considered for profiling GAS in the healthy hosts, with a view to obtaining better understanding of these organisms. The GAS-specific primers (16S rRNA and V-Na+-ATPase) used in this study can be used to estimate the maximum potential GAS carriage in people. The online version of this article (doi:10.1186/s12866-016-0858-5) contains supplementary material, which is available to authorized users.
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