An evaluation of direct PCR assays for the detection and quantification of Porphyromonas gingivalis. (2020)

• Record Type: Journal Article
• Authors: Gu, B. L.; Qi, Y. J.; Kong, J. Y.; Li, Z. T.; Wang, J. P.; Yuan, X.; Gao, S. G.
• Is Part Of: Epidemiology and infection. Volume 148(2020)
• Publication Date: 2020
• Subjects: Direct PCR, -- epidemiological investigation, -- Porphyromonas gingivalis; Communicable diseases -- Periodicals; Epidemiology -- Periodicals; 614.4
• DOI: 10.1017/S0950268820001053
• Languages: English
• ISSNs: 0950-2688

Abstract

Abstract: Porphyromonas gingivalis has been linked to the development and progression of oesophageal squamous cell carcinoma (ESCC), and is considered to be a high-risk factor for ESCC. Currently, the commonly used methods for P. gingivalis detection are culture or DNA extraction-based, which are either time and labour intensive especially for high-throughput applications. We aimed to establish and evaluate a rapid and sensitive direct quantitative polymerase chain reaction (qPCR) protocol for the detection of P. gingivalis without DNA extraction which is suitable for large-scale epidemiological studies. Paired gingival swab samples from 192 subjects undergoing general medical examinations were analysed using two direct and one extraction-based qPCR assays for P. gingivalis . Tris-EDTA buffer-based direct qPCR (TE-direct qPCR), lysis-based direct qPCR (lysis-direct qPCR) and DNA extraction-based qPCR (kit-qPCR) were used, respectively, in 192, 132 and 60 of these samples for quantification of P. gingivalis . The sensitivity and specificity of TE-direct qPCR was 95.24% and 100% compared with lysis-direct qPCR, which was 100% and 97.30% when compared with kit-qPCR; TE-direct qPCR had an almost perfect agreement with lysis-direct qPCR ( κ = 0.954) and kit-qPCR ( κ = 0.965). Moreover, the assay time used for TE-direct qPCR was 1.5 h. In conclusion, the TE-direct qPCR assay is a simple and efficient method for the quantification of oral P. gingivalis and showed high sensitivity and specificity compared with routine qPCR.