Isolation of Porphyromonas gingivalis from Esophageal Squamous Cell Carcinoma Tissues using Fastidious Anaerobe Agar-Esopageal Cancer Magazine Publishing Limited

¹ Henan Key Laboratory of Microbiome and Esophageal Cancer Prevention and Treatment; Henan Key Laboratory of Cancer Epigenetics, Cancer Hospital, The First Affiliated Hospital (College of Clinical Medicine) of Henan University of Science and Technology, Jianxi, Luoyang, China, 471003
² Diagnostic Laboratory, The First Affiliated Hospital (College of Clinical Medicine) of Henan University of Science and Technology, Jianxi, Luoyang, China, 471003
³State Key Laboratory of Oncology in South China, Collaborative Innovation Center for Cancer Medicine, Sun Yat-sen University Cancer Center, Guangzhou, China, 510060.

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Abstract:

Objective To isolate and culture P. gingivalis from esophageal squamous cell carcinoma (ESCC) tissues using fastidious anaerobe agar (FAA). Methods The effect of FAA was evaluated by assessing by assessing the growth cycle and colony characteristics of the reference strain Pg ATCC 33277. Positive P. gingivalis status was confirmed by qPCR. Tissue samples were homogenized and cultured through continuous streaking on the FAA plate in an anaerobic chamber. Preliminary identification of isolates was conducted using Real time PCR and 16S rRNA sequencing . Results FAA exhibited higher sensitivity and better suitability for the growth of P. gingivalis compared to Brain Heart Infusion Agar (BHIA) and Tryptic Soy Agar (TSA). A P. gingivalis strain (designated LyEC01) was successfully isolated from tissue of a patient with ESCC in central China. Using FAA in an anaerobic chamber, agar colonies and cultures of LyEC01 was obtained and purified. In addition, the strain was further analyzed through Sanger sequencing and mass spectrometry. Conclusion LyEC01 is a novel clinical strain of P. gingivalis, marking the first successful isolation from ESCC tissue using the FAA method.

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