title: Quantitative Analysis of Periodontal Pathogens Using Real-Time Polymerase Chain Reaction (PCR).
abstract: The quantitative polymerase chain reaction (qPCR) is a variant of PCR aimed to detect and quantify a targeted DNA molecule through the addition of probes labeled with fluorescent molecules that emit fluorescence within each amplification cycle, what results in fluorescence values proportional to the amount of accumulated PCR product. This chapter presents the detailed procedures for quantification of different periodontal pathogens (Porphyromonas gingivalis, Aggregatibacter actinomycetemcomitans, Tannerella forsythia, Campylobacter rectus, and Fusobacterium spp.) using qPCR. It also includes the description of the most frequent problems encountered and how to solve them. In addition, a detailed protocol for multiplex qPCR to detect and quantify P. gingivalis and A. actinomycetemcomitans is included.
authors.affiliation: Oral Research Laboratory, Faculty of Odontology, University Complutense, Madrid, Spain.
authors.forename: Mª José
authors.initials: MJ
authors.lastname: Marin
authors.affiliation: Oral Research Laboratory, Faculty of Odontology, University Complutense, Madrid, Spain.|Etiology and Therapy of Periodontal Diseases (ETEP) Research Group, University Complutense, Madrid, Spain.|Department of Periodontology, Faculty of Dentistry, University Complutense of Madrid, Plaza Ramón y Cajal, s/n (Ciudad Universitaria), Madrid, 28040, Spain.
authors.forename: Elena
authors.initials: E
authors.lastname: Figuero
authors.affiliation: Etiology and Therapy of Periodontal Diseases (ETEP) Research Group, University Complutense, Madrid, Spain.|Department of Periodontology, Faculty of Dentistry, University Complutense of Madrid, Plaza Ramón y Cajal, s/n (Ciudad Universitaria), Madrid, 28040, Spain.
authors.forename: David
authors.initials: D
authors.lastname: Herrera
authors.affiliation: Etiology and Therapy of Periodontal Diseases (ETEP) Research Group, University Complutense, Madrid, Spain. marsan@ucm.es.|Department of Periodontology, Faculty of Dentistry, University Complutense of Madrid, Plaza Ramón y Cajal, s/n (Ciudad Universitaria), Madrid, 28040, Spain. marsan@ucm.es.
authors.forename: Mariano
authors.initials: M
authors.lastname: Sanz
mesh_terms: D005883:Gingival Crevicular Fluid / Q000382:microbiology; D006801:Humans; D054892:Metagenome; D056186:Metagenomics / Q000379:methods; D010510:Periodontal Diseases / Q000382:microbiology*; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods
keywords: Aggregatibacter actinomycetemcomitans; Campylobacter rectus; Fusobacterium spp; Multiplex qPCR; Porphyromonas gingivalis; Primers; Probe; Quantitative PCR (qPCR); Tannerella forsythia; TaqMan