fimA genotypes and PFGE profile patterns in Porphyromonas gingivalis isolates from subjects with periodontitis.

P J Perez-Chaparro, A Rouillon, J Minet, G I Lafaurie, M Bonnaure-Mallet
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引用次数: 15

Abstract

Background and objectives: Porphyromonas gingivalis is frequently identified to type by evaluation of fimA polymorphisms and less often by pulsed-field gel electrophoresis (PFGE) because of the technical intricacies of PFGE. To compare these techniques, we genotyped P. gingivalis clinical isolates as to (i) their fimA type and (ii) their whole genome restriction profile (PFGE analysis).

Material and methods: Thirty-two P. gingivalis strains were isolated from 16 unrelated periodontitis patients. Two strains were isolated from each patient. Strains were subjected to a fimA-typing polymerase chain reaction (PCR) assay. Strains that could not be typed by PCR were submitted to sequencing of the entire fimA gene. The PFGE profiles of clinical strains were compared using bioinformatic analysis.

Results: Seven of the 32 isolates were not typeable by PCR and so their entire fimA gene was sequenced. The sequencing identified each strain as belonging to a single fimA type. In one case, sequencing of the fimA gene did not agree with the result obtained using fimA PCR typing. With the exception of one patient, each patient presented isolates bearing the same fimA type. However, in three patients, isolates with the same fimA type presented different PFGE pulsotypes.

Conclusion: The P. gingivalis typing using fimA PCR has limitations in typeability and discriminatory power. A typing technique for P. gingivalis that is easy to perform but that presents adequate typeability and discriminatory power is needed if we want to better understand the epidemiology of periodontal disease.

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牙周炎患者牙龈卟啉单胞菌分离株的fimA基因型和PFGE谱。
背景和目的:由于脉冲场凝胶电泳(PFGE)技术的复杂性,通常通过fimA多态性评估来识别牙龈卟啉单胞菌的类型,而较少使用脉冲场凝胶电泳(PFGE)。为了比较这些技术,我们根据(i)它们的fimA型和(ii)它们的全基因组限制性谱(PFGE分析)对临床分离的牙龈假单胞菌进行了基因分型。材料与方法:从16例无亲缘关系的牙周炎患者中分离到32株牙龈假单胞菌。从每位患者中分离出两株菌株。对菌株进行fima分型聚合酶链反应(PCR)检测。对无法通过PCR分型的菌株进行fimA全基因测序。采用生物信息学方法比较临床菌株的PFGE谱。结果:32株分离株中有7株无法用PCR分型,因此对其fimA基因进行了全序列测序。测序确定每个菌株属于单一的fimA型。在一个病例中,fimA基因的测序与fimA PCR分型的结果不一致。除1例患者外,所有患者的分离株均具有相同的fimA类型。然而,在3例患者中,具有相同fimA型的分离株呈现不同的PFGE脉冲型。结论:fimA PCR分型牙龈假单胞菌在分型能力和鉴别能力上存在局限性。如果我们想更好地了解牙周病的流行病学,就需要一种易于执行但具有足够的分型能力和区分能力的牙龈卟啉卟啉菌分型技术。
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