The aim is to explore the predictive value of salivary bacteria for the presence of esophageal squamous cell carcinoma (ESCC). Saliva samples were obtained from 178 patients with ESCC and 101 healthy controls

and allocated to screening and verification cohorts

respectively. In the screening phase

after saliva DNA was extracted

16S rRNA V4 regions of salivary bacteria were amplified by polymerase chain reaction (PCR) with high-throughput sequencing. Highly expressed target bacteria were screened by Operational Taxonomic Units clustering

species annotation and microbial diversity assessment. In the verification phase

the expression levels of target bacteria identified in the screening phase were verified by absolute quantitative PCR (Q-PCR). Receiver operating characteristic (ROC) curves were plotted to investigate the predictive value of target salivary bacteria. LEfSe analysis revealed higher proportions of Fusobacterium

Streptococcus and Porphyromonas

and Q-PCR assay showed significantly higher numbers of

Streptococcus salivarius

Fusobacterium nucleatum

and

Porphyromonas gingivalis

in patients with ESCC

when compared with healthy controls (all

P

<

 0.05). The areas under the ROC curves for

Streptococcus salivarius

Fusobacterium nucleatum

Porphyromonas gingivalis

and the combination of the three bacteria for predicting patients with ESCC were 69%

56.5%

61.8% and 76.4%

respectively. The sensitivities corresponding to cutoff value were 69.3%

22.7%

35.2% and 86.4%

respectively

and the matched specificity were 78.4%

96.1%

90.2% and 58.8%

respectively. These highly expressed

Streptococcus salivarius

Fusobacterium nucleatum

and

Porphyromonas gingivalis

in the saliva

alone or in combination

indicate their predictive value for ESCC.