Oral Microbiome Profiles: 16S rRNA Pyrosequencing and Microarray Assay Comparison

Columbia University, United States of America
PLoS ONE (Impact Factor: 3.23). 07/2011; 6(7):e22788. DOI: 10.1371/journal.pone.0022788
Source: PubMed


The human oral microbiome is potentially related to diverse health conditions and high-throughput technology provides the possibility of surveying microbial community structure at high resolution. We compared two oral microbiome survey methods: broad-based microbiome identification by 16S rRNA gene sequencing and targeted characterization of microbes by custom DNA microarray.
Oral wash samples were collected from 20 individuals at Memorial Sloan-Kettering Cancer Center. 16S rRNA gene survey was performed by 454 pyrosequencing of the V3-V5 region (450 bp). Targeted identification by DNA microarray was carried out with the Human Oral Microbe Identification Microarray (HOMIM). Correlations and relative abundance were compared at phylum and genus level, between 16S rRNA sequence read ratio and HOMIM hybridization intensity.
The major phyla, Firmicutes, Proteobacteria, Bacteroidetes, Actinobacteria, and Fusobacteria were identified with high correlation by the two methods (r = 0.70∼0.86). 16S rRNA gene pyrosequencing identified 77 genera and HOMIM identified 49, with 37 genera detected by both methods; more than 98% of classified bacteria were assigned in these 37 genera. Concordance by the two assays (presence/absence) and correlations were high for common genera (Streptococcus, Veillonella, Leptotrichia, Prevotella, and Haemophilus; Correlation = 0.70-0.84).
Microbiome community profiles assessed by 16S rRNA pyrosequencing and HOMIM were highly correlated at the phylum level and, when comparing the more commonly detected taxa, also at the genus level. Both methods are currently suitable for high-throughput epidemiologic investigations relating identified and more common oral microbial taxa to disease risk; yet, pyrosequencing may provide a broader spectrum of taxa identification, a distinct sequence-read record, and greater detection sensitivity.

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Available from: Zhiheng Pei, Jul 07, 2014
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    • "The phyla proportions may differ in these studies; however, the most common phyla in oral saliva are relatively constant[7,14]. Cyanobacteria are ubiquitous in water[45], and several papers have reported the presence of these bacteria in the oral cavity[7,42]. We previously proposed that these bacteria may be ingested through drinking water or by consuming contaminated seafood[45]. "
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    ABSTRACT: Recently, high-throughput sequencing has improved the understanding of the microbiological etiology of caries, but the characteristics of the microbial community structure in the human oral cavity with and without caries are not completely clear. To better understand these characteristics, Illumina MiSeq high-throughput sequencing was utilized to analyze 20 salivary samples (10 caries-free and 10 caries) from subjects from the same town in Dongxiang, Gansu, China. A total of 5,113 OTUs (Operational Taxonomic Units, 97% cutoff) were characterized in all of the salivary samples obtained from the 20 subjects. A comparison of the two groups revealed that (i) the predominant phyla were constant between the two groups; (ii) the relative abundance of the genera Veillonella, Bifidobacterium, Selenomonas, Olsenella, Parascardovia, Scardovia, Chryseobacterium, Terrimonas, Burkholderia and Sporobacter was significantly higher in the group with caries (P < 0.05); and (iii) four genera with low relative abundance (< 0.01% on average), including two characteristic genera in caries (Chryseobacterium and Scardovia), significantly influenced the microbial community structure at the genus and OTU levels. Moreover, via co-occurrence and principal component analyses, the co-prevalence of the pathogenic genera was detected in the caries samples, but in the caries-free samples, the function of clustered genera was more random. This result suggests that a synergistic effect may be influencing the assembly of the caries microbial community, whereas competition may play a more dominant role in governing the microbial community in the caries-free group. Our findings regarding the characteristics of the microbial communities of the groups with and without caries might improve the understanding of the microbiological etiology of caries and might improve the prevention and cure of caries in the future.
    Full-text · Article · Jan 2016 · PLoS ONE
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    • "The HOMIM's ability to detect 300 of the most prevalent oral bacterial species has made it a suitable method for assessing community profiles at the phylum level as well as many common taxa at the genus level. However, the HOMIM microarray method fails to detect approximately half of the bacterial species commonly present in saliva (Ahn et al., 2011). "
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    • "Although, the compatibility of the fragment length should be according with sequencing platform read length capacity. There are many studies that target different regions of the 16S rRNA gene, for example V3–V5 [29], V1–V2 [30], V1–V3 [26], V4–V5 [31], and V8–V9 [32]. There is an active discussion about the hypervariable region that should be sequenced to perform a microbial diversity analysis. "
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