Microbiota in the Throat and Risk Factors for Laryngeal Carcinoma

Gong, Hongli; Shi, Yi; Zhou, Xia; Wu, Chunping; Cao, Pengyu; Xu, Chen; Hou, Dongsheng; Wang, Yuezhu; Zhou, Liang

doi:10.1128/aem.02329-14

Cited by 65 publications

(65 citation statements)

References 48 publications

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“…6 In another study examining the microbiome of patients with laryngeal squamous cell carcinoma, the same genera were identified. 7,8 Staphylococcus was not identified in either of these studies.…”

Section: Introductionmentioning

confidence: 58%

Factors Associated With Infectious Laryngitis: A Retrospective Review of 15 Cases

Thomas

Jetté

Clary

2017

Ann Otol Rhinol Laryngol

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Objectives To identify the culturable microbes associated with infectious laryngitis and outline effective treatment strategies. Methods This is a retrospective chart review of adult patients with persistent dysphonia plus evidence of laryngeal inflammation who underwent biopsy for culture at a tertiary care medical center. Demographic factors, symptoms as reported on validated patient assessment tools, past medical history, social history, culture results, and treatment duration and response were reviewed. Results Fifteen patients with infectious laryngitis were included in this study. Culture results demonstrated Methicillin-sensitive Staphylococcus aureus (MSSA), Methicillin-resistant Staphylococcus aureus (MRSA), Pseudomonas aeruginosa, Serratia marcescens, and “normal respiratory flora.” In most patients, multiple courses of prolonged antibiotics were needed to treat MSSA or MRSA. Infections associated with other microbes resolved with a single course of antibiotics. Conclusions In this population, infectious laryngitis is defined as colonization with bacteria not found in the previously characterized laryngeal microbiome of benign vocal fold lesions. In suspected cases of infectious laryngitis, culture is recommended, by biopsy if needed. For MSSA- and MRSA-associated laryngitis, an extended course of antibiotics may be necessary for symptom improvement and resolution of laryngeal inflammation. However, the optimal treatment regimen has yet to be defined and will require larger, prospective studies.

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Section: Introductionmentioning

confidence: 58%

Factors Associated With Infectious Laryngitis: A Retrospective Review of 15 Cases

Thomas

Jetté

Clary

2017

Ann Otol Rhinol Laryngol

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“…To date, there are four published studies examining laryngeal microbiota using next-generation sequencing technology[11, 13–15], and most of these studies are limited by lack of a true control group. Further, given the paucity of data, there is no consensus on sampling methodology.…”

Section: Discussionmentioning

confidence: 99%

“…Though they reported differences in relative abundance of 15 genera in cancer versus “control”, their control group was not disease-free given the presence of benign disease, nor were the samples procured from the same anatomical site. In their second study, Gong et al[13] again compared the microbiota of laryngeal cancer specimens to vocal fold polyps, sampling from two sites using two sampling methods. Their data appear to show distinct microbial communities in swabs versus biopsies; however, they collected swabs from a different location (upper throat near epiglottis) in the larynx than where they collected tissues (vocal fold), confounding interpretation of these data.…”

Section: Discussionmentioning

confidence: 99%

“…Studying the microbiota by procuring biopsies of vocal fold tissue from live humans is not ethically feasible, as even small tissue deficits have negative effects on voice[12] and it is not currently possible to regenerate vocal fold tissue. Therefore, in the literature addressing the human laryngeal microbiome, researchers have relied on tissue sampling of mucosal biopsies adjacent to the vocal folds[11, 13] or benign vocal fold lesions[14, 15]. This limitation in sampling has impeded our ability to study the role microbes play in laryngeal health and disease.…”

Section: Introductionmentioning

confidence: 99%

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A comparison of sampling methods for examining the laryngeal microbiome

et al. 2017

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Shifts in healthy human microbial communities have now been linked to disease in numerous body sites. Noninvasive swabbing remains the sampling technique of choice in most locations; however, it is not well known if this method samples the entire community, or only those members that are easily removed from the surface. We sought to compare the communities found via swabbing and biopsied tissue in true vocal folds, a location that is difficult to sample without causing potential damage and impairment to tissue function. A secondary aim of this study was to determine if swab sampling of the false vocal folds could be used as proxy for true vocal folds. True and false vocal fold mucosal samples (swabbed and biopsied) were collected from six pigs and used for 454 pyrosequencing of the V3–V5 region of the 16S rRNA gene. Most of the alpha and beta measures of diversity were found to be significantly similar between swabbed and biopsied tissue samples. Similarly, the communities found in true and false vocal folds did not differ considerably. These results suggest that samples taken via swabs are sufficient to assess the community, and that samples taken from the false vocal folds may be used as proxies for the true vocal folds. Assessment of these techniques opens an avenue to less traumatic means to explore the role microbes play in the development of diseases of the vocal folds, and perhaps the rest of the respiratory tract.

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“…Firmicutes were analysed as this phylum is a predominant constituent of the human oral microbiome (Gong et al, 2014, Segata et al, 2012. GAPDH was analysed as a surrogate marker of human DNA contamination.…”

Section: Accepted M Manuscriptmentioning

confidence: 99%

The yield and quality of cellular and bacterial DNA extracts from human oral rinse samples are variably affected by the cell lysis methodology

Sohrabi

Nair

Samaranayake

et al. 2016

Journal of Microbiological Methods

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Q., The yield and quality of cellular and bacterial DNA extracts from human oral rinse samples are variably affected by the cell lysis methodology, Journal of Microbiological Methods (2016Methods ( ), doi: 10.1016Methods ( /j.mimet.2016 This is a PDF file of an unedited manuscript that has been accepted for publication. As a service to our customers we are providing this early version of the manuscript. The manuscript will undergo copyediting, typesetting, and review of the resulting proof before it is published in its final form. Please note that during the production process errors may be discovered which could affect the content, and all legal disclaimers that apply to the journal pertain. A C C E P T E DM A N U S C R I P T ACCEPTED MANUSCRIPT A C C E P T E D M A N U S C R I P T ACCEPTED MANUSCRIPT Highlights Lysozyme-achromopeptidase cell lysis yielded the highest gDNA level  Lysozyme-zirconium beads cell lysis extracted the highest level of bacterial DNA  Lysozyme-zirconium beads cell lysis yielded the highest level of Firmicutes A C C E P T E D M A N U S C R I P T ACCEPTED MANUSCRIPT ABSTRACTRecent culture-independent studies have enabled detailed mapping of human microbiome that has not been hitherto achievable by culture-based methods. DNA extraction is a key element of bacterial culture-independent studies that critically impacts on the outcome of the detected microbial profile. Despite the variations in DNA extraction methods described in the literature, no standardized technique is available for the purpose of microbiome profiling.Hence, standardization of DNA extraction methods is urgently needed to yield comparable data from different studies. We examined the effect of eight different cell lysis protocols on the yield and quality of the extracted DNA from oral rinse samples. These samples were exposed to cell lysis techniques based on enzymatic, mechanical, and a combination of enzymatic-mechanical methods. The outcome measures evaluated were total bacterial population, Firmicutes levels and human DNA contamination (in terms of surrogate GAPDH levels). We noted that all three parameters were significantly affected by the method of cell lysis employed. Although the highest yield of gDNA was obtained using lysozymeachromopeptidase method, the lysozyme-zirconium beads method yielded the peak quantity of total bacterial DNA and Firmicutes with a lower degree of GAPDH contamination compared with the other methods. Taken together our data clearly points to an urgent need for a consensus, standardized DNA extraction technique to evaluate the oral microbiome using oral rinse samples. Further, if Firmicutes levels are the focus of investigation in oral rinse microbiome analyses then the lysozyme-zirconium bead method would be the method of choice in preference to others.

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Microbiota in the Throat and Risk Factors for Laryngeal Carcinoma

Cited by 65 publications

References 48 publications

Factors Associated With Infectious Laryngitis: A Retrospective Review of 15 Cases

Factors Associated With Infectious Laryngitis: A Retrospective Review of 15 Cases

A comparison of sampling methods for examining the laryngeal microbiome

The yield and quality of cellular and bacterial DNA extracts from human oral rinse samples are variably affected by the cell lysis methodology

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