Detection of Coxiella burnetii from ticks by Polymerase Chain Reaction and Restriction Fragment Length Polymorphism

Çapın, Gülay Altay; Emre, Zişan; Canpolat, Seyit; Vatansever, Yusuf; Düzgün, Ali

doi:10.1501/vetfak_0000002590

Cited by 12 publications

(8 citation statements)

References 26 publications

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“…Regarding the phylogeny analysis of Coxiella sp., the OTU0019 was closely related to Coxiella-like endosymbionts (Fig 6B), previously found in R. decoloratus and R. evertsi, which have been reported as evolutionarily related but distinct to C. burnetii [49]. Because Coxiella species have been previously reported in ticks of the genera Amblyomma, Dermacentor, Ixodes, Ornitodorus and Rhipicephalus [39,50,51], it might be useful to use a multilocus sequence typing strategy in future investigations, in order to achieve the Coxiella identification in R. microplus [52]. https://doi.org/10.1371/journal.pone.0234005.g005…”

Section: Analysis Using Sequences Of the V3 Through V4 Regions Of Thesupporting

confidence: 67%

“…burnetii [ 49 ]. Because Coxiella species have been previously reported in ticks of the genera Amblyomma , Dermacentor , Ixodes , Ornitodorus and Rhipicephalus [ 39 , 50 , 51 ], it might be useful to use a multilocus sequence typing strategy in future investigations, in order to achieve the Coxiella identification in R . microplus [ 52 ].…”

Section: Resultsmentioning

confidence: 99%

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Assessment of bacterial diversity of Rhipicephalus microplus ticks from two livestock agroecosystems in Antioquia, Colombia

et al. 2020

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Rhipicephalus microplus is recognized as a tick species highly prevalent in cattle, with a wide pantropical distribution that seems to continue spreading geographically. However, its role as a biological vector has been scarcely studied in the livestock context. In this study, a 16S rRNA next-generation sequencing analysis was used to determine bacterial diversity in salivary glands and gut of R. microplus from two contrasting livestock agroecosystems in Antioquia, Colombia. Both the culture-independent approach (CI) and the culture-dependent (CD) approach were complementarily adopted in this study. A total of 341 unique OTUs were assigned, the richness showed to be higher in the Northern than in the Middle Magdalena region, and a high diversity was found at the phylum and genus levels in the samples obtained. With the CI approach, Proteobacteria, Bacteroidetes, Firmicutes, and Actinobacteria were the most common phylum of bacteria regardless of the organ, or geographic origin of the specimens analyzed. While the relative abundance of bacteria at a phylum level with the CD approach varied between analyzed samples, the data obtained suggest that a high diversity of species of bacteria occurs in R. microplus from both livestock agroecosystems. Bacterial genera such as Anaplasma, Coxiella, and Ehrlichia, recognized for their implications in tick-borne diseases, were also detected, together with endosymbionts such as Lysinibacillus, previously reported as a potential tool for biological control. This information is useful to deepen the knowledge about microbial diversity regarding the relations between endosymbionts and pathogens and could facilitate the future development of epidemiological surveillance in livestock systems.

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Section: Analysis Using Sequences Of the V3 Through V4 Regions Of Thesupporting

confidence: 67%

Section: Resultsmentioning

confidence: 99%

Assessment of bacterial diversity of Rhipicephalus microplus ticks from two livestock agroecosystems in Antioquia, Colombia

et al. 2020

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“…Polymerase chain reaction (PCR) and related techniques are widely used as sensitive and specific tools for the detection of C. burnetii in ticks, such as conventional PCR [ 16 ], restriction fragment length polymorphism (RFLP)-PCR [ 17 , 18 ], and direct sequencing [ 19 ]. The repetitive, transposon-like element, named IS1111, is a specific DNA marker for the sensitive detection of C. burnetii, in transposon (Trans)-PCR [ 5 , 20 ].…”

Section: Introductionmentioning

confidence: 99%

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

et al. 2021

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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

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“…Polymerase chain reaction (PCR) and related techniques are widely used as sensitive and speci c tools for the detection of C. burnetii in ticks, such as conventional PCR [17], restriction fragment length polymorphism (RFLP)-PCR [18,19], and direct sequencing [20]. The repetitive, transposon-like element, named IS1111, is a speci c DNA marker for sensitive detection of C. burnetii, in transposon (Trans)-PCR [5,21].…”

Section: Introductionmentioning

confidence: 99%

Real-time PCR Biochip for On-Site Detection of Coxiella Burnetii in Ticks

Truong

Yun

Lim

et al. 2021

Preprint

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Background: Q fever, a zoonosis caused by Coxiella burnetii, has adverse effects on public health. Ticks are the natural reservoirs 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 5,644 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 (p = 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 originated from goats, humans, and ticks in different countries, such as USA, France, Germany, and Serbia. Conclusions: The results of 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 for alleviating the transmission and economic damage due to Q fever.

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Detection of Coxiella burnetii from ticks by Polymerase Chain Reaction and Restriction Fragment Length Polymorphism

Cited by 12 publications

References 26 publications

Assessment of bacterial diversity of Rhipicephalus microplus ticks from two livestock agroecosystems in Antioquia, Colombia

Assessment of bacterial diversity of Rhipicephalus microplus ticks from two livestock agroecosystems in Antioquia, Colombia

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

Real-time PCR Biochip for On-Site Detection of Coxiella Burnetii in Ticks

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