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.
Keywords:
Borrelia / RT-PCR / 16S rRNA / Ixodes ricinus
Source:
Arquivo Brasileiro de Medicina Veterinaria e Zootecnia, 2010, 62, 4, 862-867
Publisher:
Arquivo Brasileiro Medicina Veterinaria Zootecnia, Minas Gerais
TY - JOUR
AU - Radulović, Željko
AU - Milutinović, Marija
AU - Tomanović, Snežana
AU - Mulenga, Albert
PY - 2010
UR - http://rimi.imi.bg.ac.rs/handle/123456789/318
AB - 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.
PB - Arquivo Brasileiro Medicina Veterinaria Zootecnia, Minas Gerais
T2 - Arquivo Brasileiro de Medicina Veterinaria e Zootecnia
T1 - Detection of Borrelia-specific 16S rRNA sequence in total RNA extracted from Ixodes ricinus ticks
EP - 867
IS - 4
SP - 862
VL - 62
DO - 10.1590/S0102-09352010000400015
ER -
@article{
author = "Radulović, Željko and Milutinović, Marija and Tomanović, Snežana and Mulenga, Albert",
year = "2010",
abstract = "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.",
publisher = "Arquivo Brasileiro Medicina Veterinaria Zootecnia, Minas Gerais",
journal = "Arquivo Brasileiro de Medicina Veterinaria e Zootecnia",
title = "Detection of Borrelia-specific 16S rRNA sequence in total RNA extracted from Ixodes ricinus ticks",
pages = "867-862",
number = "4",
volume = "62",
doi = "10.1590/S0102-09352010000400015"
}
Radulović, Ž., Milutinović, M., Tomanović, S.,& Mulenga, A.. (2010). Detection of Borrelia-specific 16S rRNA sequence in total RNA extracted from Ixodes ricinus ticks. in Arquivo Brasileiro de Medicina Veterinaria e Zootecnia
Arquivo Brasileiro Medicina Veterinaria Zootecnia, Minas Gerais., 62(4), 862-867.
https://doi.org/10.1590/S0102-09352010000400015
Radulović Ž, Milutinović M, Tomanović S, Mulenga A. Detection of Borrelia-specific 16S rRNA sequence in total RNA extracted from Ixodes ricinus ticks. in Arquivo Brasileiro de Medicina Veterinaria e Zootecnia. 2010;62(4):862-867.
doi:10.1590/S0102-09352010000400015 .
Radulović, Željko, Milutinović, Marija, Tomanović, Snežana, Mulenga, Albert, "Detection of Borrelia-specific 16S rRNA sequence in total RNA extracted from Ixodes ricinus ticks" in Arquivo Brasileiro de Medicina Veterinaria e Zootecnia, 62, no. 4 (2010):862-867,
https://doi.org/10.1590/S0102-09352010000400015 . .
