Rapid quantification of periodontitis-related bacteria using a novel modification of Invader PLUS technologies

Abstract

The Invader PLUS technology is a sensitive, rapid method for the detection and quantification of nucleic acid. While the original technology is based on the amplification by polymerase chain reaction (PCR) of the target sequence followed by its detection using the Invader technology, the current modification allows simultaneous PCR amplification and Invader reaction. The PCR primers and the Invader probes are designed to operate at the same temperature. This allows simpler design and faster results. This technology has been applied for the quantification of six periodontitis-related bacteria (Actinobacillus actinomycetemcomitans, Porphyromonas gingivalis, Prevotella intermedia, Toreponema denticola, Tannerella forsythensis and Fusobacterium nucleatum). Direct comparison of this modified Invader PLUS with real-time PCR demonstrated similar linear range. Furthermore, testing of 64 volunteers showed a good correlation between both technologies with correlation factors r2 spanning between 0.827 and 0.987. We demonstrated here that the proposed improvement of the Invader PLUS allows the detection and quantification of DNA sequences using a simple design and protocol that can be implemented in clinical testing.

The rs8099917 Polymorphism, When Determined by a Suitable Genotyping Method, Is a Better Predictor for Response to Pegylated Alpha Interferon/Ribavirin Therapy in Japanese Patients than Other Single Nucleotide Polymorphisms Associated with Interleukin-28B

“…on August 28, 2018 by guest http://jcm.asm.org/ sitivity was the greatest of the three methods (HP, TaqMan, and Invader), as reported previously (20). Genotyping error for rs8099917 by direct sequencing due to novel SNP.…”

Quantification of Periodontopathic Bacteria in Saliva Using the Invader Assay

“…Tadokoro et al showed that the Invader Plus method could quantify periodontal pathogens with good correlation with the results of real-time PCR (19). In this assay, DNA sequences were amplified and quantified using the Invader technology.…”

The evaluation of bacterial flora in progress of peri‐implant disease

“…15 Bacterial DNA was purified using MagNA Pure LC (Roche, Basel, Switzerland) and added to a 15-ll reaction mixture containing 50 lM d-NTP, 700 nM primer probe, 70 nM Invader oligo, and 2.5 U AmpliTaq Stoffel fragment (Applied Biosystems, Foster City, CA, USA), and the Cleavase XI Invader core re-agent kit (Genomic DNA, Third Wave Technologies Inc., Madison, WI, USA) consisting of Cleavase XI FRET mix and Cleavase XI enzyme ⁄ MgCl 2 solution. The reaction mixture was preheated at 95°C for 2 minutes, followed by 35 cycles of two-step PCR (95°C for 1 minute, 63°C for 1 minute) using LightCycler 480 (Roche, Basel, Switzerland).…”