Differentiation of banding patterns between Streptococcus mutans and Streptococcus sobrinus isolates in rep-PCR using ERIC primer.
Differentiation of banding patterns between Streptococcus mutans and Streptococcus sobrinus isolates in rep-PCR using ERIC primer.
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DOI：
10.3402/jom.v3i0.7190
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发表时间：
2011
影响因子：
4.5
通讯作者：
Michalek SM
中科院分区：
文献类型：
--
作者：
Okada T;Takada K;Fujita K;Ikemi T;Osgood RC;Childers NK;Michalek SM
变形链球菌和远缘链球菌被认为是人类龋病发生的重要菌种。因此，建立一种可靠的基因分型方法来区分沙门氏菌。变形链球菌S. sobrinus是至关重要的。我们评估了使用ERIC引物带型的重复基因外回文聚合酶链反应（rep-PCR）在区分S。mutans和S. sobrinus。五S变形链球菌和两株S. sobrinus原型菌株和50株临床分离株（38株;变形链球菌C型4株; sobrinus血清型d和8个S. sobrinus血清型g）。比较原型菌株和临床分离株扩增产物的带型，寻找区分沙门氏菌的共同带型。mutans和S. sobrinus。所有供试菌株均观察到多条带型。对S.变异株在2，000 bp、1，700 bp、1，400 bp、1，100 bp、850 bp和250 bp处显示6条独特的强条带; sobrinus在2,000 bp、1,800 bp、1,100 bp、900 bp、800 bp、600 bp和550 bp处有7条强条带。在1,100 bp处的条带是在两个S. mutans和S. sobrinus。此外，大多数临床S.变异株显示相同的带型。所有鼠伤寒沙门变形链球菌在1,700 bp、850 bp和250 bp处有扩增产物，而变形链球菌在1,700 bp、850 bp和250 bp处有扩增产物。sobrinus的核苷酸序列分别为1，100 bp、900 bp和800 bp。这些结果表明，使用ERIC引物的rep-PCR可以区分S。mutans和S. sobrinus。
Streptococcus mutans and Streptococcus sobrinus are considered to be important bacterial species in the initiation of human dental caries. Therefore, the establishment of a reliable genotyping method to distinguish S. mutans from S. sobrinus is of central importance. We assessed the usefulness of repetitive extragenic palindromic polymerase chain reaction (rep-PCR) using ERIC primer banding patterns in differentiating S. mutans and S. sobrinus. Five S. mutans and two S. sobrinus prototype strains and 50 clinical isolates (38 S. mutans serotype c, 4 S. sobrinus serotype d, and 8 S. sobrinus serotype g) were examined. The banding patterns of amplicons generated were compared among the prototype strains and clinical isolates, to find common bands that distinguish S. mutans and S. sobrinus. Multiple banding patterns were seen with all strains tested. The representative strains of S. mutans tested revealed six unique, strong bands at 2,000 bp, 1,700 bp, 1,400 bp, 1,100 bp, 850 bp, and 250 bp, whereas S. sobrinus had seven strong bands at 2,000 bp, 1,800 bp, 1,100 bp, 900 bp, 800 bp, 600 bp, and 550 bp. The band at 1,100 bp was the only band that was observed in both S. mutans and S. sobrinus. Furthermore, most clinical S. mutans isolates revealed identical banding patterns. All S. mutans had amplicons at 1,700 bp, 850 bp, and 250 bp, whereas those of S. sobrinus were at 1,100 bp, 900 bp, and 800 bp. These results indicate that using rep-PCR with the ERIC primers can distinguish between S. mutans and S. sobrinus.