Metagenomic analysis to identify the causative agents of atypical urogenital tract infections
- Authors: Kutilin D.S.1
-
Affiliations:
- National Medical Research Oncology Center, Rostov-on-Don, Russian Federation
- Issue: Vol 11, No 6 (2021)
- Pages: 1108-1122
- Section: ORIGINAL ARTICLES
- Submitted: 30.03.2021
- Accepted: 12.09.2021
- Published: 11.10.2021
- URL: https://iimmun.ru/iimm/article/view/1713
- DOI: https://doi.org/10.15789/2220-7619-MAT-1713
- ID: 1713
Cite item
Full Text
Abstract
Background. Research in recent decades has shown that classical microbiological methods can only detect a small, cultivable portion of microorganisms. One of the modern approaches to detect a wide range of bacteria and archaea is presented as a type of metagenomic analysis performed by high-throughput sequencing of ribosomal operon fragment libraries. Objective is to conduct metagenomic analysis of samples from the urogenital tract of patients with chronic inflammation to identify pathogens not detected by other methods. Methods. Taxonomic analysis of the bacterial community was performed by high-throughput sequencing of the V3–V4 hypervariable region of the 16S rRNA gene by using the Illumina HiSeq 3000 platform. Results. The study allowed to identify the taxonomic diversity of microorganisms in samples from the urogenital tract (from 197 to 794 different microorganisms belonging to the Bacteria were identified), as well as to establish differential differences concerning members of the genera Megasphaera, Prevotella, Veillonella, Pedobacter, Mobiluncus, Phobormidium, Sphing Temperatibacter, Oxobacter, Georgenia, Actinobaculum, Varibaculum, Mycobacterium, Rhodococcus, Sediminihabitans, Actinobacter, Actinoplanes, Spirochaeta, Enhydrobacter, Thermacetogenium, Bdellovibrio, Oleibacter, Porphyromonas, Klebsiella, Lachnoclostridium, Caulobacter, Xanthomonas, Novispirillum, Marvinbryantia, Afipia, Shinella, Tepidimonas, Faecalibacterium, Paludibacterium, Aerococcus, Campylobacter, Pasteurella, Rumen, Psychrobacter, Haemophilus, Brevibacillus, Sporosarcina, Yaniella and Lactobacillus between samples from patients with chronic inflammation and apparently healthy individuals. Conclusion. Differential differences were found in the composition of the microbiome from the samples of patients with chronic inflammation and apparently healthy individuals, concerning members of 44 genera, including Megasphaera, Prevotella, Veillonella, Pedobacter, Mobiluncus, Phormidium and Lactobacillus. The inflammatory processes observed in the patient urogenital tract can be associated with imbalanced microflora such as decreased level of typical members of the genera Staphylococcus, Streptococcus and Lactobacillus, but increased number of members from of the genera Klebsiella and Citrobacter.
About the authors
D. S. Kutilin
National Medical Research Oncology Center, Rostov-on-Don, Russian Federation
Author for correspondence.
Email: k.denees@yandex.ru
ORCID iD: 0000-0002-8942-3733
Denis S. Kutilin - PhD (Biology), Leading Researcher, Laboratory of Molecular Oncology, National Medical Research Oncology Center, Rostov-on-Don, Russian Federation.
344037, Rostov-on-Don, Liniya 14 str., 63.
Phone: +7 951 538-50-39
РоссияReferences
- Карпеева Ю.С., Новикова В.П., Хавкин А.И., Ковтун Т.А., Макаркин Д.В., Федотова О.Б. Микробиота и болезни человека: возможности диетической коррекции. Российский вестник перинатологии и педиатрии. 2020. T. 65, № 5. С. 116–125. doi: 10.21508/1027-4065-2020-65-5-116-125
- Кунгуров Н.В., Герасимова Н.М., Горбунов А.П., Скидан Н.И., Щербакова Н.В., Евстигнеева Н.П., Чигвинцева Е.А., Юровских Л.И., Тамбулова В.Н. Условно-патогенная микрофлора урогенитального тракта у мужчин с воспалительными заболеваниями мочеиспускательного канала. Вестник последипломного медицинского образования. 2010. № 2. С. 26–32.
- Ткачук В.Н. Хронический простатит. М.: Медицина для всех, 2006. 112 с.
- Anderson-Otunu O., Akhtar S. Chronic infections of the urinary tract and bladder cancer risk: a systematic review. Asian Pac. J. Cancer Prev., 2016, vol. 17, no. 8, pp. 3805–3807.
- Aragón I.M., Herrera-Imbroda B., Queipo-Ortuño M.I., Castillo E., Del Moral J.S., Gómez-Millán J., Yucel G., Lara M.F. The urinary tract microbiome in health and disease. Eur. Urol. Focus, 2018, vol. 4, no. 1, pp. 128–138. doi: 10.1016/j.euf.2016.11.001
- Bates S.T., Berg-Lyons D., Caporaso J.G., Walters W.A., Knight R., Fierer N. Examining the global distribution of dominant archaeal populations in soil. ISME J., 2010, vol. 5, pp. 908–917.
- Bolger A.M., Lohse M., Usadel B. Trimmomatic: a flexible trimmer for Illumina sequence data. Bioinformatics, 2014, vol. 30, no. 15, pp. 2114–2120. doi: 10.1093/bioinformatics/btu170
- Bordigoni A., Lo C.I., Yimagou E.K., Diop K., Nicaise B., Raoult D., Desnues C., Fenollar F. Megasphaera vaginalis sp. nov. and Anaerococcus vaginimassiliensis sp. nov., new bacteria isolated from vagina of French woman with bacterial vaginosis. New Microbes New Infect., 2020, vol. 37: 100706. doi: 10.1016/j.nmni.2020.100706
- Caporaso J.G., Kuczynski J., Stombaugh J., Bittinger K., Bushman F.D., Costello E.K., Fierer N., Peña A.G., Goodrich J.K., Gordon J.I., Huttley G.A., Kelley S.T., Knights D., Koenig J.E., Ley R.E., Lozupone C.A., McDonald D., Muegge B.D., Pirrung M., Reeder J., Sevinsky J.R., Turnbaugh P.J., Walters W.A., Widmann J., Yatsunenko T., Zaneveld J., Knight R. QIIME allows analysis of high-throughput community sequencing data. Nat. Methods., 2010, vol. 7, no. 5, pp. 335–336. doi: 10.1038/nmeth.f.303
- Drewes J.L., White J.R., Dejea C.M., Fathi P., Iyadorai T., Vadivelu J., Roslani A.C., Wick E.C., Mongodin E.F., Loke M.F., Thulasi K., Gan H.M., Goh K.L., Chong H.Y., Kumar S., Wanyiri J.W., Sears C.L. High-resolution bacterial 16S rRNA gene profile meta-analysis and biofilm status reveal common colorectal cancer consortia. NPJ Biofilms Microbiomes, 2017, vol. 3: 34. doi: 10.1038/s41522-017-0040-3
- Edgar R.C., Haas B.J., Clemente J.C., Quince C., Knight R. UCHIME improves sensitivity and speed of chimera detection. Bioinformatics, 2011, vol. 27, no. 16, pp. 2194–2200. doi: 10.1093/bioinformatics/btr381
- Fadrosh D.W., Ma B., Gajer P., Sengamalay N., Ott S., Brotman R.M., Ravel J. An improved dual-indexing approach for multi-plexed 16S rRNA gene sequencing on the Illumina MiSeq platform. Microbiome, 2014, vol. 2, no. 1: 6. doi: 10.1186/2049-2618-2-6
- Gilbert J.A., Blaser M.J., Caporaso J.G., Jansson J.K., Lynch S.V., Knight R. Current understanding of the human microbiome. Nat. Med., 2018, vol. 24, no. 4, pp. 392–400. doi: 10.1038/nm.4517
- Huang C.H., Chou Y.H., Yeh H.W., Huang J.Y., Yang S.F., Yeh C.B. Risk of cancer after lower urinary tract infection: a population-based cohort study. Int. J. Environ Res. Public Health, 2019, vol. 16, no. 3: 390. doi: 10.3390/ijerph16030390
- Jackobson S.H., Eklof O., Eriksson C.G., Lins L.E., Tidgren B., Winberg J. Development of hypertension and ureamia after pyelonephritis in childhood: 27 year follow up. BMJ, 1989, vol. 299, no. 6701, pp. 703–706. doi: 10.1136/bmj.299.6701.703
- Jensen E.A., Berryman D.E., Murphy E.R., Carroll R.K., Busken J., List E.O., Broach W.H. Heterogeneity spacers in 16S rDNA primers improve analysis of mouse gut microbiomes via greater nucleotide diversity. Biotechniques, 2019, vol. 67, no. 2, pp. 55–62. doi: 10.2144/btn-2019-0025
- Kim M.S., Jung S.I. The urinary tract microbiome in male genitourinary diseases: focusing on benign prostate hyperplasia and lower urinary tract symptoms. Int. Neurourol. J., 2021, vol. 25, no. 1, pp. 3–11. doi: 10.5213/inj.2040174.087
- Langmead B., Salzberg S.L. Fast gapped-read alignment with Bowtie 2. Nat. Methods, 2012, vol. 9, no. 4, pp. 357–359. doi: 10.1038/nmeth.1923
- Mändar R., Punab M., Korrovits P., Türk S., Ausmees K., Lapp E., Preem J.K., Oopkaup K., Salumets A., Truu J. Seminal microbiome in men with and without prostatitis. Int. J. Urol., 2017, vol. 24, no. 3, pp. 211–216. doi: 10.1111/iju.13286
- Masajtis-Zagajewska A., Nowicki M. New markers of urinary tract infection. Clin. Chim. Acta, 2017, vol. 471, pp. 286–291. doi: 10.1016/j.cca.2017.06.003
- Moustafa A., Li W., Singh H., Moncera K.J., Torralba M.G., Yu Y., Manuel O., Biggs W., Venter J.C., Nelson K.E., Pieper R., Telenti A. Microbial metagenome of urinary tract infection. Sci. Rep., 2018, vol. 8, no. 1: 4333. doi: 10.1038/s41598-018-22660-8
- Neugent M.L., Hulyalkar N.V., Nguyen V.H., Zimmern P.E., De Nisco N.J. Advances in understanding the human urinary microbiome and its potential role in urinary tract infection. mBio, 2020, vol. 11, no. 2: e00218-20. doi: 10.1128/mBio.00218-20
- Randis T.M., Ratner A.J. Gardnerella and Prevotella: co-conspirators in the pathogenesis of bacterial vaginosis. J. Infect. Dis., 2019, vol. 220, no. 7, pp. 1085–1088. doi: 10.1093/infdis/jiy705
- Scheiman J., Luber J.M., Chavkin T.A., MacDonald T., Tung, A., Pham L.-D., Wibowo M.C., Wurth R.C., Punthambaker S., Tierney B.T., Yang Z., Hattab M.W., Avila-Pacheco J., Clish C.B., Lessard S., Church G.M., Kostic A.D. Meta-omics analysis of elite athletes identifies a performance-enhancing microbe that functions via lactate metabolism. Nat. Med., 2019, vol. 25, no. 7, pp. 1104–1109. doi: 10.1038/s41591-019-0485-4
- Shrestha E., White J.R., Yu S.H., Kulac I., Ertunc O., De Marzo A.M., Yegnasubramanian S., Mangold L.A., Partin A.W., Sfanos K.S. Profiling the urinary microbiome in men with positive versus negative biopsies for prostate Cancer. J. Urol., 2018, vol. 199, no. 1, pp. 161–171. doi: 10.1016/j.juro.2017.08.001
- Steen A.D., Crits-Christoph A., Carini P., DeAngelis K.M., Fierer N., Lloyd K.G., Thrash J.C. High proportions of bacteria and archaea across most biomes remain uncultured. ISME J., 2019, vol. 13, pp. 3126–3130.
- Srinivasan S., Chambers L.C., Tapia K.A., Hoffman N.G., Munch M.M., Morgan J.L., Domogala D., Lowens M.S., Proll S., Huang M.-L., Soge O.O., Jerome K.R., Golden M.R., Hughes J.P., Fredricks D.N., Manhart L.E. Urethral microbiota in men: association of Haemophilus influenzae and mycoplasma penetrans with nongonococcal urethritis. Clin. Infect. Dis., 2021, vol. 73, no. 7, pp. e1684–e1693. doi: 10.1093/cid/ciaa1123
- Tanaka S., Yoshida M., Murakami Y., Ogiwara T., Shoji M., Kobayashi S., Watanabe S., Machino M., Fujisawa S. The relationship of Prevotella intermedia, Prevotella nigrescens and Prevotella melaninogenica in the supragingival plaque of children, caries and oral malodor. J. Clin. Pediatr. Dent., 2008, vol. 32, no. 3, pp. 195–200. doi: 10.17796/jcpd.32.3.vp657177815618l1
- Wang Q., Garrity G.M., Tiedje J.M., Cole J.R. Naive Bayesian classifier for rapid assignment of rRNA sequences into the new bacterial taxonomy. Appl. Environ. Microbiol., 2007, vol. 73, no. 16, pp. 5261–5267. doi: 10.1128/AEM.00062-07