Cocolonization of Pneumococcal Serotypes in Healthy Children Attending Day Care Centers: Molecular Versus Conventional Methods
Cocolonization of Pneumococcal Serotypes in Healthy Children Attending Day Care Centers: Molecular Versus Conventional Methods
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DOI：
10.1097/inf.0000000000001059
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发表时间：
2016-05-01
影响因子：
3.6
通讯作者：
Haraldsson, Gunnsteinn
中科院分区：
文献类型：
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作者：
Hjalmarsdottir, Martha A.;Gudmundsdottir, Palina Fanney;Haraldsson, Gunnsteinn
目的：肺炎球菌是常见的定植菌，尤其是儿童，不同血清型的共定植是种内遗传交换的重要因素。本研究的目的是分析在冰岛未接种疫苗的健康儿童的肺炎球菌携带和血清型分布，并比较传统的培养方法和分子方法，使用DNA提取直接从samples.Methods：鼻咽拭子从514名2-6岁的儿童参加日托中心在雷克雅未克在2009年。选择性培养拭子中的肺炎球菌，并使用乳胶凝集法对分离株进行血清分型。DNA也提取直接从拭子和血清分型使用多重PCR面板设计检测疫苗血清型和最常见的非疫苗serotypes.Result：肺炎球菌携带检测391（76.1%）的儿童使用聚合酶链反应（PCR）和371（72.2%）使用常规方法。PCR法检出共定植菌92例（23.5%），常规法检出共定植菌30例（8.1%），分别检出500株和401株（P < 0.0001）。最常见的血清型为23 F、19 A、6 B、6A和19 F，占13- 8%。10价疫苗和13价疫苗所含血清型的PCR检出菌株数为234株（58.4%）和363（90.5%）和常规方法186（46.4%）和293（73.1%）。使用直接从鼻咽拭子中提取的DNA，在四分之一携带肺炎球菌的儿童中检测到共定植。携带率很高，但没有血清型占优势，儿童普遍被疫苗血清型定植，尤其是共定植儿童。
Objectives: Pneumococci are common colonizer, especially of children, and cocolonization of different serotypes is an important factor for intraspecies genetic exchange. The aim of this study was to analyze pneumococcal carriage and serotype distribution in unvaccinated healthy children in Iceland and compare conventional culture methods and molecular methods using DNA extracted directly from the samples.Methods: Nasopharyngeal swabs were obtained from 514 children aged 2-6 year attending day care centers in Reykjavik in 2009. The swabs were selectively cultured for pneumococci and the isolates serotyped using latex agglutination. DNA was also extracted directly from the swabs and serotyped using a multiplex PCR panel designed to detect vaccine serotypes and the most commonly carried non-vaccine serotypes.Result: Pneumococcal carriage was detected in 391 (76.1%) of the children using polymerase chain reaction (PCR) and in 371 (72.2%) using conventional methods. Cocolonization was detected in 92 (23.5%) of the carriers when PCR method was used and in 30 (8.1%) when conventional methods were used, detecting 500 and 401 strains, respectively (P < 0.0001). The most common serotypes were 23F, 19A, 6B, 6A and 19F, rates 13-8%. The number of isolates of serotypes included in the 10-valent and 13-valent vaccines and detected by PCR were 234 (58.4%) and 363 (90.5%), respectively and by conventional methods 186 (46.4%) and 293 (73.1%), respectively.Conclusion: Cocolonization was detected in a fourth of the children carrying pneumococci using DNA extracted directly from nasopharyngeal swabs. The rate of carriage was very high, but no serotype dominated, and the children were commonly colonized by vaccine serotypes, especially cocolonized children.