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Real-time PCR biochip for on-site detection of Coxiella burnetii in ticks

BACKGROUND: Q fever, a zoonosis caused by Coxiella burnetii, has adverse effects on public health. Ticks are vectors of C. burnetii and they contribute to the transmission of the pathogen. A tool for rapid, sensitive, and accurate detection of C. burnetii from ticks is important for the prevention o...

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Autores principales: Truong, A.-Tai, Yun, Bo-Ram, Lim, Jiyeon, Min, Subin, Yoo, Mi-Sun, Yoon, Soon-Seek, Yun, Young-Min, Kim, Jong-Taek, Cho, Yun Sang
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8101159/
https://www.ncbi.nlm.nih.gov/pubmed/33957987
http://dx.doi.org/10.1186/s13071-021-04744-z
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author Truong, A.-Tai
Yun, Bo-Ram
Lim, Jiyeon
Min, Subin
Yoo, Mi-Sun
Yoon, Soon-Seek
Yun, Young-Min
Kim, Jong-Taek
Cho, Yun Sang
author_facet Truong, A.-Tai
Yun, Bo-Ram
Lim, Jiyeon
Min, Subin
Yoo, Mi-Sun
Yoon, Soon-Seek
Yun, Young-Min
Kim, Jong-Taek
Cho, Yun Sang
author_sort Truong, A.-Tai
collection PubMed
description BACKGROUND: Q fever, a zoonosis caused by Coxiella burnetii, has adverse effects on public health. Ticks are vectors of C. burnetii and they contribute to the transmission of the pathogen. A tool for rapid, sensitive, and accurate detection of C. burnetii from ticks is important for the prevention of Q fever. METHODS: Ultra-rapid real-time PCR (UR-qPCR) as a chip-based real-time PCR system was developed for the detection of C. burnetii from ticks. The UR-qPCR system was established and evaluated for the rapidity, sensitivity, and specificity of C. burnetii detection. RESULTS: C. burnetii was detected using UR-qPCR from 5644 larval, nymphal, and adult ticks from 408 pools collected from livestock and epidemiologically linked environments in two provinces, Gangwon and Jeju, in Korea. Ticks from three species were identified; Haemaphysalis longicornis accounted for the highest number, present in 333 of 408 pools (81.62%), followed by Haemaphysalis flava in 62 pools (15.19%) and Ixodes nipponensis in 13 pools (3.19%). The rapidity and sensitivity of PCR detection was demonstrated with the sufficient amplification and detection of approximately 56 copies of C. burnetii DNA with only 20 min of PCR amplification. The kappa value for the diagnostic agreement between UR-qPCR and stationary qPCR was in perfect agreement (κ = 1). PCR detection and sequencing indicated that C. burnetii was present in 5 of the 408 pools (1.23%), in which four pools contained H. longicornis and one pool contained H. flava. The infection rates of C. burnetii in the tick pools collected from Gangwon and Jeju Provinces were 1.70% and 0.58%, respectively. Phylogenetic analysis indicated a close relationship between the detected C. burnetii and those originating from goats, humans, and ticks in different countries, such as the USA, France, Germany, and Serbia. CONCLUSIONS: The methods described in this study could be important for the prevention and control of Q fever in the two provinces. The UR-qPCR, with its features of mobility, sensitivity, and rapidity, is helpful for constructing early alert systems in the field for C. burnetii in ticks and could help alleviate the transmission of and economic damage due to Q fever. GRAPHIC ABSTRACT: [Image: see text] SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s13071-021-04744-z.
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spelling pubmed-81011592021-05-06 Real-time PCR biochip for on-site detection of Coxiella burnetii in ticks Truong, A.-Tai Yun, Bo-Ram Lim, Jiyeon Min, Subin Yoo, Mi-Sun Yoon, Soon-Seek Yun, Young-Min Kim, Jong-Taek Cho, Yun Sang Parasit Vectors Research BACKGROUND: Q fever, a zoonosis caused by Coxiella burnetii, has adverse effects on public health. Ticks are vectors of C. burnetii and they contribute to the transmission of the pathogen. A tool for rapid, sensitive, and accurate detection of C. burnetii from ticks is important for the prevention of Q fever. METHODS: Ultra-rapid real-time PCR (UR-qPCR) as a chip-based real-time PCR system was developed for the detection of C. burnetii from ticks. The UR-qPCR system was established and evaluated for the rapidity, sensitivity, and specificity of C. burnetii detection. RESULTS: C. burnetii was detected using UR-qPCR from 5644 larval, nymphal, and adult ticks from 408 pools collected from livestock and epidemiologically linked environments in two provinces, Gangwon and Jeju, in Korea. Ticks from three species were identified; Haemaphysalis longicornis accounted for the highest number, present in 333 of 408 pools (81.62%), followed by Haemaphysalis flava in 62 pools (15.19%) and Ixodes nipponensis in 13 pools (3.19%). The rapidity and sensitivity of PCR detection was demonstrated with the sufficient amplification and detection of approximately 56 copies of C. burnetii DNA with only 20 min of PCR amplification. The kappa value for the diagnostic agreement between UR-qPCR and stationary qPCR was in perfect agreement (κ = 1). PCR detection and sequencing indicated that C. burnetii was present in 5 of the 408 pools (1.23%), in which four pools contained H. longicornis and one pool contained H. flava. The infection rates of C. burnetii in the tick pools collected from Gangwon and Jeju Provinces were 1.70% and 0.58%, respectively. Phylogenetic analysis indicated a close relationship between the detected C. burnetii and those originating from goats, humans, and ticks in different countries, such as the USA, France, Germany, and Serbia. CONCLUSIONS: The methods described in this study could be important for the prevention and control of Q fever in the two provinces. The UR-qPCR, with its features of mobility, sensitivity, and rapidity, is helpful for constructing early alert systems in the field for C. burnetii in ticks and could help alleviate the transmission of and economic damage due to Q fever. GRAPHIC ABSTRACT: [Image: see text] SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s13071-021-04744-z. BioMed Central 2021-05-06 /pmc/articles/PMC8101159/ /pubmed/33957987 http://dx.doi.org/10.1186/s13071-021-04744-z Text en © The Author(s) 2021 https://creativecommons.org/licenses/by/4.0/Open AccessThis article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) . The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/ (https://creativecommons.org/publicdomain/zero/1.0/) ) applies to the data made available in this article, unless otherwise stated in a credit line to the data.
spellingShingle Research
Truong, A.-Tai
Yun, Bo-Ram
Lim, Jiyeon
Min, Subin
Yoo, Mi-Sun
Yoon, Soon-Seek
Yun, Young-Min
Kim, Jong-Taek
Cho, Yun Sang
Real-time PCR biochip for on-site detection of Coxiella burnetii in ticks
title Real-time PCR biochip for on-site detection of Coxiella burnetii in ticks
title_full Real-time PCR biochip for on-site detection of Coxiella burnetii in ticks
title_fullStr Real-time PCR biochip for on-site detection of Coxiella burnetii in ticks
title_full_unstemmed Real-time PCR biochip for on-site detection of Coxiella burnetii in ticks
title_short Real-time PCR biochip for on-site detection of Coxiella burnetii in ticks
title_sort real-time pcr biochip for on-site detection of coxiella burnetii in ticks
topic Research
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8101159/
https://www.ncbi.nlm.nih.gov/pubmed/33957987
http://dx.doi.org/10.1186/s13071-021-04744-z
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