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首页> 外文期刊>Journal of Clinical Microbiology >Development of a TaqMan Array Card for Pneumococcal Serotyping on Isolates and Nasopharyngeal Samples
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Development of a TaqMan Array Card for Pneumococcal Serotyping on Isolates and Nasopharyngeal Samples

机译：TaqMan阵列卡在分离株和鼻咽样品上进行肺炎球菌血清分型的开发

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Streptococcus pneumoniae is both a commensal and a major pathogen that causes invasive disease in people of all ages. The introduction of serotype-specific pneumococcal vaccines has reduced the burden of disease but has also led to replacement with new strains; thus, serotyping remains important for vaccine-related disease surveillance. Conventional serotyping methods are laborious and expensive. We developed an easy-to-perform genotypic TaqMan array card (TAC) to identify S. pneumoniae strains, including lytA-based sequences, and 53 sequence-specific PCRs to identify 74 serotypes/serogroups covering all current vaccine types as well as prevalent nonvaccine types. The TAC method was evaluated on 146 clinical S. pneumoniae isolates and 13 nonpneumococcal species that naturally inhabit the upper respiratory tract and yielded 97% (142/146) sensitivity and 100% (13/13) specificity versus results of standard Quellung serotyping. The calculated limit of detection was 20 to 200 fg (～8 to 84 genome equivalents) per reaction. On 23 blinded nasopharyngeal specimens that were pneumococcus culture positive, the TAC pan-pneumococcus lytA assay was positive in 21 (91% sensitivity versus culture). On TAC lytA-positive specimens, a serotype result was obtained on 86%, and the result was 95% accurate versus the subsequent culture's Quellung result. TAC also detected mixed serotypes in two specimens where Quellung detected only the predominant serotype. This TAC method yields fast and comprehensive serotyping compared to the standard method and may be useful on direct specimens.
机译：肺炎链球菌既是常见病又是引起所有年龄段人群浸润性疾病的主要病原体。血清型特异性肺炎球菌疫苗的引入减轻了疾病负担，但也导致了新菌株的替代。因此，血清分型对于疫苗相关疾病的监测仍然很重要。常规的分型方法费力且昂贵。我们开发了易于执行的基因型TaqMan阵列卡（TAC）来鉴定肺炎链球菌菌株，包括基于 lytA 的序列，并进行53种序列特异性PCR来鉴定涵盖所有当前流行病的74种血清型/血清型疫苗类型以及流行的非疫苗类型。使用TAC方法对自然栖息于上呼吸道的146种临床肺炎链球菌和13种非肺炎球菌物种进行了评估，相对于标准Quellung血清分型的结果，其产生97％（142/146）的敏感性和100％（13/13）的特异性。每个反应的计算检出限为20至200 fg（约8至84个基因组当量）。在23例肺炎球菌培养呈阳性的盲鼻咽标本上，TAC泛肺炎球菌 lytA 测定在21例中呈阳性（相对于培养，敏感性为91％）。在TAC lytA 阳性标本上，获得了86％的血清型结果，与后续培养的Quellung结果相比，该结果的准确性为95％。 TAC还检测了两个标本中的混合血清型，而Quellung仅检测到了主要血清型。与标准方法相比，这种TAC方法可快速，全面地进行血清分型，可能对直接样本有用。

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