Fukui K, Kato N, Kato H, Watanabe K, Tatematsu N
Institute of Anaerobic Bacteriology, Gifu University School of Medicine, Gifu 500-8705, Japan.
J Clin Microbiol. 1999 Oct;37(10):3141-5. doi: 10.1128/JCM.37.10.3141-3145.1999.
We established a typing system for Prevotella intermedia and Prevotella nigrescens using the combination of PCR ribotyping and arbitrarily primed PCR (AP-PCR) fingerprinting and applied this system to the study of intrafamilial incidence of these species in the oral cavity. PCR ribotyping followed by subtyping by AP-PCR fingerprinting was applied to each type strain of P. intermedia and P. nigrescens and 54 isolates (32 isolates of P. intermedia and 24 isolates of P. nigrescens) from extraoral infections, resulting in an excellent discriminatory power (discrimination index, 0.99) for both species. A total of 18 subjects from six families, with the subjects from each family comprising the mother, the father, and a child who had subclinical early-stage to moderate adult periodontitis or simple gingivitis and who carried P. intermedia or P. nigrescens, or both, were enrolled in the study of intrafamilial carriage. When 20 colonies per specimen of subgingival plaque, if available, were picked from primary culture, 115 P. intermedia and 178 P. nigrescens isolates were recovered from the 18 subjects. Among the subjects studied, family members shared the same subtype strain(s) but non-family members did not. Multiple subtypes were found in 8 (57%) of the 14 P. nigrescens-positive subjects but in only 3 (27%) of the 11 P. intermedia-positive subjects; the difference was, however, not statistically significant (P = 0.14). These results suggest that the combination of PCR ribotyping and AP-PCR fingerprinting is well suited for the epidemiological study of P. intermedia and P. nigrescens and that each family seems to carry a distinct subtype(s) of these species.
我们采用聚合酶链反应核糖体分型(PCR ribotyping)和任意引物聚合酶链反应(AP-PCR)指纹图谱相结合的方法,建立了中间普氏菌(Prevotella intermedia)和变黑普氏菌(Prevotella nigrescens)的分型系统,并将该系统应用于研究这些菌种在口腔内的家族内感染发生率。对中间普氏菌和变黑普氏菌的每一株标准菌株,以及从口外感染中分离出的54株菌株(32株中间普氏菌和24株变黑普氏菌)进行了先PCR核糖体分型,再通过AP-PCR指纹图谱进行亚型分型,结果显示这两种菌种均具有出色的鉴别能力(鉴别指数为0.99)。共有来自六个家庭的18名受试者参与了家族内携带情况的研究,每个家庭的受试者包括母亲、父亲以及患有亚临床早期至中度成人牙周炎或单纯牙龈炎且携带中间普氏菌或变黑普氏菌或两者皆有的儿童。若有龈下菌斑标本,从原代培养物中每标本挑选20个菌落,从这18名受试者中分离出115株中间普氏菌和178株变黑普氏菌。在所研究的受试者中,家庭成员共享相同的亚型菌株,而非家庭成员则没有。在14名变黑普氏菌阳性受试者中有8名(57%)发现了多种亚型,但在11名中间普氏菌阳性受试者中只有3名(27%)发现了多种亚型;然而,差异无统计学意义(P = 0.14)。这些结果表明,PCR核糖体分型和AP-PCR指纹图谱相结合的方法非常适合用于中间普氏菌和变黑普氏菌的流行病学研究,并且每个家庭似乎携带这些菌种的独特亚型。