Igarashi T, Yamamoto A, Goto N
Department of Oral Microbiology, Showa University School of Dentistry, Tokyo, Japan.
Oral Microbiol Immunol. 1996 Oct;11(5):294-8. doi: 10.1111/j.1399-302x.1996.tb00184.x.
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.
变形链球菌是人类龋齿的病原体。已开发出一种通过聚合酶链反应直接从人类牙菌斑中检测变形链球菌的方法。设计了针对变形链球菌英布里特株(血清型c)葡聚糖酶基因(dexA)一部分的寡核苷酸引物,通过聚合酶链反应扩增出1272bp的DNA片段。本方法能特异性检测变形链球菌(血清型c、e和f),但不能检测其他变形链球菌:仓鼠链球菌(血清型a)、鼠链球菌(血清型b)、远缘链球菌(血清型d和g)和唐氏链球菌(血清型h),也不能检测其他革兰氏阳性菌(12种球菌的16株和12种杆菌的18株)及革兰氏阴性菌(1种球菌的1株和18种杆菌的20株)。该方法能够检测从变形链球菌英布里特株纯化的1pg染色体DNA以及少至12个变形链球菌细胞的菌落形成单位。人类牙菌斑中的变形链球菌细胞也能被直接检测到。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经聚合酶链反应检测均为阳性。这些结果表明,dexA聚合酶链反应适用于变形链球菌的特异性检测和鉴定。