Detection of Borrelia-specific 16S rRNA sequence in total RNA extracted from Ixodes ricinus ticks
A reverse transcriptase - polymerase chain reaction based assay for Borrelia species detection in ticks was developed. The method was based on amplification of 552 nucleotide bases long sequence of 16S rRNA, targeted by Borrelia specific primers. In the present study, total RNA extracted from Ixodes...
| Main Authors: | , , , |
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| Format: | Article |
| Language: | English |
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Universidade Federal de Minas Gerais
2010-08-01
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| Series: | Arquivo Brasileiro de Medicina Veterinária e Zootecnia |
| Subjects: | |
| Online Access: | http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0102-09352010000400015&lng=en&tlng=en |
| _version_ | 1828762978747416576 |
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| author | Ž. Radulović M. Milutinović S. Tomanović A. Mulenga |
| author_facet | Ž. Radulović M. Milutinović S. Tomanović A. Mulenga |
| author_sort | Ž. Radulović |
| collection | DOAJ |
| description | A reverse transcriptase - polymerase chain reaction based assay for Borrelia species detection in ticks was developed. The method was based on amplification of 552 nucleotide bases long sequence of 16S rRNA, targeted by Borrelia specific primers. In the present study, total RNA extracted from Ixodes ricinus ticks was used as template. The results showed higher sensitivity for Borrelia detection as compared to standard dark-field microscopy. Method specificity was confirmed by cloning and sequencing of obtained 552 base pairs long amplicons. Phylogenetic analysis of obtained sequences showed that they belong to B. lusitaniae and B. afzelii genospecies. RT-PCR based method presented in this paper could be very useful as a screening test for detecting pathogen presence, especially when in investigations is required extraction of total RNA from ticks. |
| first_indexed | 2024-12-11T01:53:21Z |
| format | Article |
| id | doaj.art-3209f7a6006e4b9ca92b6a84c33f6f4b |
| institution | Directory Open Access Journal |
| issn | 1678-4162 |
| language | English |
| last_indexed | 2024-12-11T01:53:21Z |
| publishDate | 2010-08-01 |
| publisher | Universidade Federal de Minas Gerais |
| record_format | Article |
| series | Arquivo Brasileiro de Medicina Veterinária e Zootecnia |
| spelling | doaj.art-3209f7a6006e4b9ca92b6a84c33f6f4b2022-12-22T01:24:41ZengUniversidade Federal de Minas GeraisArquivo Brasileiro de Medicina Veterinária e Zootecnia1678-41622010-08-0162486286710.1590/S0102-09352010000400015S0102-09352010000400015Detection of Borrelia-specific 16S rRNA sequence in total RNA extracted from Ixodes ricinus ticksŽ. Radulović0M. Milutinović1S. Tomanović2A. MulengaUniversity of BelgradeUniversity of BelgradeTexas A&M UniversityA reverse transcriptase - polymerase chain reaction based assay for Borrelia species detection in ticks was developed. The method was based on amplification of 552 nucleotide bases long sequence of 16S rRNA, targeted by Borrelia specific primers. In the present study, total RNA extracted from Ixodes ricinus ticks was used as template. The results showed higher sensitivity for Borrelia detection as compared to standard dark-field microscopy. Method specificity was confirmed by cloning and sequencing of obtained 552 base pairs long amplicons. Phylogenetic analysis of obtained sequences showed that they belong to B. lusitaniae and B. afzelii genospecies. RT-PCR based method presented in this paper could be very useful as a screening test for detecting pathogen presence, especially when in investigations is required extraction of total RNA from ticks.http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0102-09352010000400015&lng=en&tlng=enBorreliaRT-PCR16S rRNAIxodes ricinus |
| spellingShingle | Ž. Radulović M. Milutinović S. Tomanović A. Mulenga Detection of Borrelia-specific 16S rRNA sequence in total RNA extracted from Ixodes ricinus ticks Arquivo Brasileiro de Medicina Veterinária e Zootecnia Borrelia RT-PCR 16S rRNA Ixodes ricinus |
| title | Detection of Borrelia-specific 16S rRNA sequence in total RNA extracted from Ixodes ricinus ticks |
| title_full | Detection of Borrelia-specific 16S rRNA sequence in total RNA extracted from Ixodes ricinus ticks |
| title_fullStr | Detection of Borrelia-specific 16S rRNA sequence in total RNA extracted from Ixodes ricinus ticks |
| title_full_unstemmed | Detection of Borrelia-specific 16S rRNA sequence in total RNA extracted from Ixodes ricinus ticks |
| title_short | Detection of Borrelia-specific 16S rRNA sequence in total RNA extracted from Ixodes ricinus ticks |
| title_sort | detection of borrelia specific 16s rrna sequence in total rna extracted from ixodes ricinus ticks |
| topic | Borrelia RT-PCR 16S rRNA Ixodes ricinus |
| url | http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0102-09352010000400015&lng=en&tlng=en |
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