The Ability of a Novel Trypsin-like Peptidase Activity Assay Kit to Detect Red-Complex Species

Abstract

The trypsin-like peptidase activity assay kit measures the trypsin-like protease produced by three red-complex species, Porphyromonas gingivalis, Tannerella forsythia, and Treponema denticola, causing periodontitis, and detects the presence of these bacteria in samples. The purpose of this study was to investigate the relationship between the detection of TLPs by a novel TLP-AA, ADCHECK and the detection of red-complex pathogens by real-time PCR using tongue swabs from patients with periodontitis. The detection limit of trypsin-like protease activity by ADCHECK was validated using the culture supernatants of two different Porphyromonas gingivalis bacterial strains. Real-time PCR was performed to determine the number of red-complex species in the tongue coatings of patients with periodontal disease. Trypsin-like protease activity in tongue-swab samples was scored using ADCHECK. ADCHECK successfully detected trypsin-like protease activity in 103 Porphyromonas gingivalis bacterial strains. The specificity, positive predictive value, negative predictive value, and accuracy of ADCHECK for the presence of red-complex pathogens determined by real-time PCR were 90%, 97%, 98%, and 92%, respectively. ADCHECK is an effective tool for the detection of red-complex pathogens.

The association between trypsin‐like protease activity in the oral cavity and kidney function in Japanese workers

“…The a* value did not show a strong correlation with clinical periodontal parameters, including mean PPD and CAL. ADCHECK ® quantifies the TLP activity in the tongue swab, and TLP is an enzyme produced by anaerobic periodontal bacteria (Usui et al, 2022). Deep periodontal pockets where anaerobic bacteria are predominant may be responsible for the high TLP activity measurement results.…”

“…The observed associations between the a * value and kidney function appear to be biologically plausible. A larger a * value is an indicator of poorer periodontal conditions with the presence of TLPs produced by red complex periodontal pathogens (Iwasaki et al, 2021; Usui et al, 2022). The response to periodontal pathogenic bacteria leads to a local tissue destruction and inflammation, which could be a local source of inflammatory cytokines and bacterial components from the periodontium.…”

“…This a * value was used in the current analyses. Greater TLP activity, as shown by ADCHECK®, was shown to be associated with increasing bacterial numbers (Usui et al, 2022).…”

“…We have demonstrated the ability of ADCHECK® to detect TLP activity produced by Porphyromonas gingivalis cultured bacterial strains, with the colour reaction in the matrix disc of ADCHECK® progressing with increasing number of bacteria. In addition, the detection accuracy of ADCHECK® for TLP present in periodontitis patient samples was demonstrated to be high when compared with the results obtained by real‐time PCR as a gold standard (Usui et al, 2022). Furthermore, the performance of the kit to detect clinically defined periodontitis has been validated (Iwasaki et al, 2021).…”

“…However, conducting clinical examinations remains challenging because of the required resources, including time, equipment and personnel (trained examiners) (Kimura et al, 2001; Yamamoto et al, 2009). Recently, an assay kit to measure the trypsin‐like protease (TLP) produced by three red‐complex bacteria, Porphyromonas gingivalis , Tannerella forsythia and Treponema denticola , was made commercially available in Japan: ADCHECK® (ADTEC Co., Ltd., USA, Japan) (Usui et al, 2022). Greater TLP activity has been associated with increased virulence of these periodontitis‐associated bacteria (Fletcher et al, 1995).…”