Isolation of Mycoplasma genitalium from patients with urogenital infections: first report from the Latin-American region
Isolation of Mycoplasma genitalium from clinical specimens remains difficult. We describe a modified culture system based on Vero cells grown in medium 199 with 2% foetal bovine serum (FBS). The culture system was evaluated using early passage M. genitalium strains M6271 and M6311 with growth monito...
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| Format: | Online |
| Language: | English |
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BlackWell Publishing Ltd
2013
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| Online Access: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4184696/ |
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pubmed-4184696 |
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pubmed-41846962014-10-29 Isolation of Mycoplasma genitalium from patients with urogenital infections: first report from the Latin-American region Mondeja, B A Jensen, J S Rodríguez, I Morier, L F Kourí, V Rodríguez, N M Fernández, C Original Articles Isolation of Mycoplasma genitalium from clinical specimens remains difficult. We describe a modified culture system based on Vero cells grown in medium 199 with 2% foetal bovine serum (FBS). The culture system was evaluated using early passage M. genitalium strains M6271 and M6311 with growth monitoring by quantitative TaqMan PCR. Eleven endocervical swabs and one male urethral swab positive by 16S rRNA and MgPa1–3 PCRs were quantified and inoculated into Vero cell suspensions in medium 199 supplemented with 2% FBS and antibiotics. Cultures were incubated for 14 days. Cell passages and growth monitoring by TaqMan PCR were performed until the growth of M. genitalium reached ≥106 geq/mL. Confirmation of the new M. genitalium strains was performed by sequencing a 281 bp fragment of mgpB. The growth of Mycoplasma genitalium strains was recorded for all urogenital swab specimens in the modified cell-culture system. Growth of M. genitalium was obtained within 2 months and yielded 12 M. genitalium strains with all 11 isolates from females of an identical, but unique genotype. To our knowledge, this is the first successful isolation of M. genitalium in the Latin-American region. The use of Vero cell culture in 199 medium with 2% FBS is a method comparable to the Ultroser G culture system for isolation of M. genitalium. Genotyping of clinical samples and isolates should be performed to document the absence of cross-contamination. BlackWell Publishing Ltd 2013-11 2013-11-28 /pmc/articles/PMC4184696/ /pubmed/25356322 http://dx.doi.org/10.1002/2052-2975.20 Text en © 2013 The Authors. New Microbes and New Infections is published by John Wiley & Sons Ltd on behalf of the European Society of Clinical Microbiology and Infectious Disease. http://creativecommons.org/licenses/by-nc/3.0/ This is an open access article under the terms of the Creative Commons Attribution-NonCommercial License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited and is not used for commercial purposes. |
| repository_type |
Open Access Journal |
| institution_category |
Foreign Institution |
| institution |
US National Center for Biotechnology Information |
| building |
NCBI PubMed |
| collection |
Online Access |
| language |
English |
| format |
Online |
| author |
Mondeja, B A Jensen, J S Rodríguez, I Morier, L F Kourí, V Rodríguez, N M Fernández, C |
| spellingShingle |
Mondeja, B A Jensen, J S Rodríguez, I Morier, L F Kourí, V Rodríguez, N M Fernández, C Isolation of Mycoplasma genitalium from patients with urogenital infections: first report from the Latin-American region |
| author_facet |
Mondeja, B A Jensen, J S Rodríguez, I Morier, L F Kourí, V Rodríguez, N M Fernández, C |
| author_sort |
Mondeja, B A |
| title |
Isolation of Mycoplasma genitalium from patients with urogenital infections: first report from the Latin-American region |
| title_short |
Isolation of Mycoplasma genitalium from patients with urogenital infections: first report from the Latin-American region |
| title_full |
Isolation of Mycoplasma genitalium from patients with urogenital infections: first report from the Latin-American region |
| title_fullStr |
Isolation of Mycoplasma genitalium from patients with urogenital infections: first report from the Latin-American region |
| title_full_unstemmed |
Isolation of Mycoplasma genitalium from patients with urogenital infections: first report from the Latin-American region |
| title_sort |
isolation of mycoplasma genitalium from patients with urogenital infections: first report from the latin-american region |
| description |
Isolation of Mycoplasma genitalium from clinical specimens remains difficult. We describe a modified culture system based on Vero cells grown in medium 199 with 2% foetal bovine serum (FBS). The culture system was evaluated using early passage M. genitalium strains M6271 and M6311 with growth monitoring by quantitative TaqMan PCR. Eleven endocervical swabs and one male urethral swab positive by 16S rRNA and MgPa1–3 PCRs were quantified and inoculated into Vero cell suspensions in medium 199 supplemented with 2% FBS and antibiotics. Cultures were incubated for 14 days. Cell passages and growth monitoring by TaqMan PCR were performed until the growth of M. genitalium reached ≥106 geq/mL. Confirmation of the new M. genitalium strains was performed by sequencing a 281 bp fragment of mgpB. The growth of Mycoplasma genitalium strains was recorded for all urogenital swab specimens in the modified cell-culture system. Growth of M. genitalium was obtained within 2 months and yielded 12 M. genitalium strains with all 11 isolates from females of an identical, but unique genotype. To our knowledge, this is the first successful isolation of M. genitalium in the Latin-American region. The use of Vero cell culture in 199 medium with 2% FBS is a method comparable to the Ultroser G culture system for isolation of M. genitalium. Genotyping of clinical samples and isolates should be performed to document the absence of cross-contamination. |
| publisher |
BlackWell Publishing Ltd |
| publishDate |
2013 |
| url |
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4184696/ |
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1613140775048052736 |