Direct detection of Streptococcus mutans in human dental plaque by polymerase chain reaction
Direct detection of Streptococcus mutans in human dental plaque by polymerase chain reaction
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DOI：
10.1111/j.1399-302x.1996.tb00184.x
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发表时间：
1996-10-01
影响因子：
--
通讯作者：
Goto, N
中科院分区：
文献类型：
--
作者：
Igarashi, T;Yamamoto, A;Goto, N
变形链球菌是人类龋齿的病原体。已经开发出一种通过聚合酶链反应直接从人牙菌斑中检测变形链球菌的方法。设计对变形链球菌 Ingbritt（血清型 c）的葡聚糖酶基因 (dexA) 部分具有特异性的寡核苷酸引物，通过聚合酶链式反应扩增 1272 bp DNA 片段。本方法特异性检测出变形链球菌（血清型 c、e 和 f），但未检测到其他变形链球菌：仓鼠链球菌（a 血清型）、鼠链球菌（b 血清型）、sobrinus（d 和 g 血清型）和 Downei 链球菌（h 血清型）以及其他革兰氏阳性菌（12 种球菌中的 16 株和 18 株链球菌）。 12种 杆菌）或革兰氏阴性菌（1 种球菌中的 1 株和 18 种杆菌中的 20 株）。该方法能够检测从变形链球菌 Ingbritt 纯化的 1 pg 染色体 DNA 和少至 12 个变形链球菌细胞的集落形成单位。还直接检测到人牙菌斑中的变形链球菌细胞。从 8 名患者牙菌斑中分离出的 70 株变形链球菌临床分离株，聚合酶链反应均呈阳性。这些结果表明dexA聚合酶链式反应适用于变形链球菌的特异性检测和鉴定。
Streptococcus mutans is an etiological agent in human dental caries. A method for the detection of S. mutans directly from human dental plaque by polymerase chain reaction has been developed. Oligonucleotide primers specific for a portion of the dextranase gene (dexA) of S. mutans Ingbritt (serotype c) were designed to amplify a 1272-bp DNA fragment by polymerase chain reaction. The present method specifically detected S. mutans (serotypes c, e and f), but none of the other mutans streptococci: S. cricetus (serotype a), S. rattus (serotype b), S. sobrinus (serotypes d and g), and S. downei (serotype h), other gram-positive bacteria(16 strains of 12 species of cocci and 18 strains of 12 species of bacilli) nor gram-negative bacteria (1 strain of 1 species of cocci and 20 strains of 18 species of bacilli). The method was capable of detecting 1 pg of the chromosomal DNA purified from S. mutans Ingbritt and as few as 12 colony-forming units of S. mutans cells. The S. mutans cells in human dental plaque were also directly detected. Seventy clinical isolates of S. mutans isolated from the dental plaque of 8 patients were all positive by the polymerase chain reaction. These results suggest that the dexA polymerase chain reaction is suitable for the specific detection and identification of S. mutans.