Angulo, A F

Galama, J M

Melchers, W J

Niesters, H G

Quint, W G

van der Logt, J T

van Kuppeveld, F J

van Zoest, M J

English

Dissertations of the University of Groningen

   University of GroningenGenus- and species-specific identification of mycoplasmas by 16S rRNA amplificationvan Kuppeveld, F J; van der Logt, J T; Angulo, A F; van Zoest, M J; Quint, W G; Niesters, HG; Galama, J M; Melchers, W JPublished in:Applied and environmental microbiologyIMPORTANT NOTE: You are advised to consult the publisher's version (publisher's PDF) if you wish to cite fromit. Please check the document version below.Document VersionPublisher's PDF, also known as Version of recordPublication date:1993Link to publication in University of Groningen/UMCG research databaseCitation for published version (APA):van Kuppeveld, F. J., van der Logt, J. T., Angulo, A. F., van Zoest, M. J., Quint, W. G., Niesters, H. G.,Galama, J. M., & Melchers, W. J. (1993). Genus- and species-specific identification of mycoplasmas by16S rRNA amplification. Applied and environmental microbiology, 59(2), 655.CopyrightOther than for strictly personal use, it is not permitted to download or to forward/distribute the text or part of it without the consent of theauthor(s) and/or copyright holder(s), unless the work is under an open content license (like Creative Commons).The publication may also be distributed here under the terms of Article 25fa of the Dutch Copyright Act, indicated by the “Taverne” license.More information can be found on the University of Groningen website: https://www.rug.nl/library/open-access/self-archiving-pure/taverne-amendment.Take-down policyIf you believe that this document breaches copyright please contact us providing details, and we will remove access to the work immediatelyand investigate your claim.Downloaded from the University of Groningen/UMCG research database (Pure): http://www.rug.nl/research/portal. For technical reasons thenumber of authors shown on this cover page is limited to 10 maximum.Download date: 21-01-2023AUTHOR'S CORRECTIONGenus- and Species-Specific Identification of Mycoplasmasby 16S rRNA AmplificationF. J. M. vAN KUPPEVELD, J. T. M. VAN DER LOGT, A. F. ANGULO, M. J. VAN ZOEST, W. G. V. QUINT,H. G. M. NIESTERS, J. M. D. GALAMA, AND W. J. G. MELCHERSDepartment of Medical Microbiology, University of Nijmegen, P.O. Box 9101, 6500 HB Nijmegen, NationalInstitute of Public Health and Environmental Protection, 3720 BA Bilthoven, and DepartmentofMolecular Biology, Diagnostic Centre SSDZ, 2600 GA Delft, The NetherlandsVolume 58, no. 8, p. 2606-2615. In this paper, we have described a mycoplasma genus-specific primer set (primers GPO-1and MGSO), which specifically amplifies a 715-bp fragment with all mycoplasmal species investigated but not with otherspecies. The specificity of this primer set, as is demonstrated in Fig. 7 in our article, was proven on the rDNA level.Very recently we have found that when the mycoplasma genus-specific primer set is used for the amplification of rRNAsequences, predominantly a 350-bp product is formed instead of the 715-bp product. Since this unusual phenomenon isobserved only on the rRNA level, not on the rDNA level (as can be seen in Fig. 7), it is most probably due to some unusualfeatures or (length) restrictions of the reverse transcription step.To avoid this problem, we have replaced primer GPO-1 with a new 5' primer (GPO-3), which is closer to the 3' primerMGSO. We have demonstrated that amplification with primer GPO-3 in conjuction with primer MGSO results in a polymerasechain reaction product of 270 bp on both the rDNA and rRNA level (see Fig. 1 in our article) and that these primers displaythe same specificity as was described previously for the primers GPO-1 and MGSO (see Fig. 2 in our article). For confirmationof the polymerase chain reaction with the primers GPO-3 and MGSO, we have used probe GPO-4.Thus, for amplification of rRNA sequences with the genus-specific primer set, we advise replacement of 5' primer GPO-1and probe GPO-2 with the following oligonucleotides, GPO-3 (new 5' primer) and GPO-4 (new probe).GPO-3 5'-GGGAGCAAACAGGATTAGATACCCT-3'IUB E. coli 16S rRNA positions 774-798GPO-4 5'-CTTAAAGGAATTGACGGGAACCCG-3'IUB E. coli 16S rRNA positions 910-933655 on October 27, 2015 by University of Groningenhttp://aem.asm.org/Downloaded from 

Genus- and species-specific identification of mycoplasmas by 16S rRNA amplification

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Genus- and species-specific identification of mycoplasmas by 16S rRNA amplification

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