Multiplex PCR for identification of six clinically relevant streptococci
Multiplex PCR for identification of six clinically relevant streptococci
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DOI：
10.1099/jmm.0.000615
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发表时间：
2017-11-01
影响因子：
3
通讯作者：
Kerdsin, Anusak
中科院分区：
文献类型：
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作者：
Hatrongjit, Rujirat;Akeda, Yukihiro;Kerdsin, Anusak
目的.本研究的目的是建立一种多重PCR（mPCR）方法，用于同时检测（单反应）6种临床相关链球菌：肺炎链球菌、猪链球菌、解没食子链球菌亚种和猪链球菌。gallolyticus、S.解没食子酸亚种巴氏链球菌、中间链球菌和咽峡炎链球菌/星座链球菌。用S. pneumoniae（lytA）、S. suis（recN）、S.解没食子酸亚种gallolyticus（tanB）、S.解没食子酸亚种pasteurianus（SGPB 0680细胞壁表面蛋白）、S. intermedius（ily）和S.使用来自泰国东北部七家三级医院的37株参考菌株和442株临床链球菌分离株进行了心绞痛/星座链球菌（moaC）的检测。将该mPCR的结果与API 20 Strep和常规生化试验的结果进行比较。6种临床相关链球菌的预期扩增产物分别为229、362、531、723、819和978 bp。肺炎链球菌（S.解没食子酸亚种gallolyticus、S.解没食子酸亚种巴氏杆菌S. suis，S. intermedius和S.心绞痛/S.星座，分别。未观察到其他供试菌种的非特异性反应。对于442株临床链球菌，该mPCR检测方法确认了菌种的同一性，与API 20 Strep和常规生化检测结果一致。该方法可用于6株链球菌纯培养物的快速鉴定。结果表明，该方法快速、简便、可靠，可用于在种属水平上鉴定这些链球菌。该检测方法可用于实验室鉴定和监测这些细菌对人类的感染。
Purpose. The aim of this study was to develop a multiplex PCR (mPCR) for simultaneous detection (single reaction) of six clinically relevant streptococcal species: Streptococcus pneumoniae, Streptococcus suis, Streptococcus gallolyticus subsp. gallolyticus, S. gallolyticus subsp. pasteurianus, Streptococcus intermedius and Streptococcus anginosus/constellatus.Methods. mPCR with primers specific for S. pneumoniae (lytA), S. suis (recN), S. gallolyticus subsp. gallolyticus (tanB), S. gallolyticus subsp. pasteurianus (SGPB0680 cell wall surface protein), S. intermedius (ily) and S. anginosus/constellatus (moaC) was employed with 37 reference bacterial strains and 442 clinical streptococcal isolates collected from seven tertiary hospitals in north-east Thailand. Results from this mPCR were compared to those obtained with the API 20 Strep and conventional biochemical tests.Results. The six clinically relevant streptococcal species gave the expected amplification products of 229, 362, 531, 723, 819 and 978 bp for S. pneumoniae, S. gallolyticus subsp. gallolyticus, S. gallolyticus subsp. pasteurianus, S. suis, S. intermedius and S. anginosus/S. constellatus, respectively. Non-specific reactions were not observed with the other bacterial species tested. For the 442 clinical streptococci, this mPCR assay confirmed the identity of the species in accordance with results obtained with the API 20 Strep and conventional biochemical tests.Conclusion. This mPCR can be applied to the rapid identification of pure cultures of these six streptococci. The test was shown to be rapid, simple and reliable for the identification of these streptococci at the species level. This assay should be useful for laboratory identification and surveillance of human infections by these bacterial species.