当前位置： SCI文献检索 > JOURNAL OF CLINICAL MICROBIOLOGY期刊下所有文献 > Highly specific protease-based approach for detection of porphyromonas gingivalis in diagnosis of periodontitis.

Highly specific protease-based approach for detection of porphyromonas gingivalis in diagnosis of periodontitis.

Abstract：

:Porphyromonas gingivalis is associated with the development of periodontitis. Here we describe the development of a highly specific protease-based diagnostic method for the detection of P. gingivalis in gingival crevicular fluid. Screening of a proteolytic peptide substrate library, including fluorogenic dipeptides that contain d-amino acids, led to the discovery of five P. gingivalis-specific substrates. Due to the presence of lysine and arginine residues in these substrates, it was hypothesized that the cleavage was mediated by the gingipains, a group of P. gingivalis-specific proteases. This hypothesis was confirmed by the observation that P. gingivalis gingipain knockout strains demonstrated clearly impaired substrate cleavage efficacy. Further, proteolytic activity on the substrates was increased by the addition of the gingipain stimulators dithiothreitol and l-cysteine and decreased by the inhibitors leupeptin and N-ethylmaleimide. Screening of saliva and gingival crevicular fluid of periodontitis patients and healthy controls showed the potential of the substrates to diagnose the presence of P. gingivalis proteases. By using paper points, a sensitivity of approximately 10(5) CFU/ml was achieved. P. gingivalis-reactive substrates fully composed of l-amino acids and Bz-l-Arg-NHPhNO(2) showed a relatively low specificity (44 to 85%). However, the five P. gingivalis-specific substrates that each contained a single d-amino acid showed high specificity (96 to 100%). This observation underlines the importance of the presence of d-amino acids in substrates used for the detection of bacterial proteases. We envisage that these substrates may improve the specificity of the current enzyme-based diagnosis of periodontitis associated with P. gingivalis.

journal_name

J Clin Microbiol

journal_title

Journal of clinical microbiology

authors

Kaman WE,Galassi F,de Soet JJ,Bizzarro S,Loos BG,Veerman EC,van Belkum A,Hays JP,Bikker FJ

doi

10.1128/JCM.05313-11

subject

Has Abstract

pub_date

2012-01-01 00:00:00

pages

104-12

issue

eissn

0095-1137

issn

1098-660X

pii

JCM.05313-11

journal_volume

pub_type

杂志文章

在线工具

Highly specific protease-based approach for detection of porphyromonas gingivalis in diagnosis of periodontitis.