An Alternative and Rapid Method for the Extraction of Nucleic Acids from Ixodid Ticks by Potassium Acetate Procedure

Rodríguez, Islay; Fraga, Jorge; Noda, Angel A.; Mayet, Milagro; Duarte, Yanisia; Echevarría, Eduardo; Fernández, Carmen

doi:10.1590/s1516-8913201402005

Cited by 17 publications

(3 citation statements)

References 22 publications

(10 reference statements)

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“…The DNA was extracted using the potassium acetate method [ 9 ]. Briefly, pooled ticks were homogenized in liquid nitrogen and sterile PBS, washed again in 70% ethanol, rinsed with sterile water, and dried.…”

Section: Methodsmentioning

confidence: 99%

Diversity of Rickettsia species in collected ticks from Southeast Iran

Qorbani,

Khalili,

Nourollahifard

et al. 2024

BMC Vet Res

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Rickettsia occurs worldwide and rickettsiosis is recognized as an emerging infection in several parts of the world. Ticks are reservoir hosts for pathogenic Rickettsia species in humans and domestic animals. Most pathogenic Rickettsia species belong to the spotted Fever Group (SFG). This study aimed to identify and diagnose tick fauna and investigate the prevalence of Rickettsia spp. in ticks collected from domestic animals and dogs in the rural regions of Kerman Province, Southeast Iran. In this study, tick species (fauna) were identified and 2100 ticks (350 pooled samples) from two genera and species including Rhipicephalus linnaei (1128) and Hyalomma deteritum (972) were tested to detect Rickettsia genus using Real-time PCR. The presence of the Rickettsia genus was observed in 24.9% (95%CI 20.28–29.52) of the pooled samples. Sequencing and phylogenetic analyses revealed the presence of Rickettsia aeschlimannii (48.98%), Rickettsia conorii israelensis (28.57%), Rickettsia sibirica (20.41%), and Rickettsia helvetica (2.04%) in the positive samples. The results showed a significant association between county variables and the following variables: tick spp. (p < 0.001), Rickettsia genus infection in ticks (p < 0.001) and Rickettsia spp. (p < 0.001). In addition, there was a significant association between tick species and host animals (dogs and domestic animals) (p < 0.001), Rickettsia spp infection in ticks (p < 0.001), and Rickettsia spp. (p < 0.001). This study indicates a high prevalence of Rickettsia spp. (SFG) in ticks of domestic animals and dogs in rural areas of Kerman Province. The health system should be informed of the possibility of rickettsiosis and the circulating species of Rickettsia in these areas.

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Section: Methodsmentioning

confidence: 99%

Diversity of Rickettsia species in collected ticks from Southeast Iran

Qorbani,

Khalili,

Nourollahifard

et al. 2024

BMC Vet Res

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“…The samples were ground using steel balls in a laboratory ball mill to isolate fungal DNA. Next, the potassium acetate method was used for DNA isolation and PCR analysis (Rodríguez et al 2014).…”

Section: Isolation and Molecular Identification Of T Aestivum Fungal ...mentioning

confidence: 99%

First Report on Truffle-Inhabiting Fungi and Metagenomic Communities of Tuber Aestivum Collected in Russia

Malygina,

Imidoeva,

Morgunova

et al. 2024

J Exp Bio & Ag Sci

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Truffles are one of the least studied groups of fungi in terms of their biological and biotechnological aspects. This study aimed to isolate truffle-inhabiting fungi and assess the metagenomic communities of the most common Russian summer truffle, Tuber aestivum. This study is the first to characterize the biodiversity of prokaryotic and eukaryotic organisms living in the truffle T. aestivum using molecular analysis and sequencing. Plant pathogens involved in a symbiotic relationship with truffles were identified by sequencing the hypervariable fragments of the 16S rRNA and 18S rRNA genes. In addition, some strains of fungal symbionts and likely pathogens were isolated and recognized for the first time from the truffles. This study also compared and characterized the general diversity and distribution of microbial taxa of T. aestivum collected in Russia and Europe. The results revealed that the Russian and European truffle study materials demonstrated high similarity. In addition to the truffles, representatives of bacteria, fungi, and protists were found in the fruiting bodies. Many of these prokaryotic and eukaryotic species inhabiting truffles might influence them, help them form mycorrhizae with trees, and regulate biological processes. Thus, truffles are interesting and promising sources for modern biotechnological and agricultural studies.

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“…Порівняно з другим методом та іншими ефективними вже описаними методиками, третій метод особливо зручний для використання при великій кількості досліджуваних зразків, а також для зразків невеликого розміру, таких як німфи, а також личинки кліщів (Mtambo et al, 2006;Rodríguez et al, 2014).…”

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Comparison of methods of DNA extraction from ixodid ticks

Levytska

Mushinsky

Dwużnik

et al. 2020

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Ixodid ticks are ectoparasites of animals and humans that carry a wide variety of pathogenic microorganisms. Ticks can cause pathological conditions such as paralysis, fever, toxicosis and allergies, as well as a large number of infectious and invasive diseases. The aim of our study was to compare three methods of DNA extraction, to test their effectiveness and practicality in obtaining material from ticks and to determine their effect on the results of PCR-based studies. During 2018, questing ticks were collected from vegetation in Khmelnytsky region. Three different methods were used for DNA extraction: crushing of ticks with scissors and lysis in ammonium hydroxide, crushing with scissors followed by DNA extraction with a commercial kit Genomic Mini AX Tissue Spin (A&A Biotechnology, Gdynia, Poland), homogenization of ticks with programmable cryogenic grinder SPEX Sample Prep Freezer Mill 6875 followed by extraction of DNA with Genomic Mini AX Tissue Spin. A total of 72 ticks (60 D. reticulatus and 12 I. ricinus) from vegetation were examined by PCR for the presence of pathogens from genera/complex Babesia, Rickettsia and Borrelia burgdorferi sensu lato (s.l.). Following the first method of DNA extraction, 4.2% of ticks tested positive for Babesia spp., and DNA of Rickettsia spp. and Borrelia burgdorferi s.l. was detected in 12.5% and 33.3% of ticks, respectively. Following the second method of DNA extraction, 4.2% of ticks tested positive for Babesia spp., and DNA of Rickettsia spp. and Borrelia burgdorferi s.l. was detected in 54.2% (13 positive specimens) and 33.3% of ticks, respectively. The third method also revealed 4.2% of ticks positive for DNA of Babesia spp. Rickettsia DNA was detected in 7 D. reticulatus and 2 I. ricinus ticks, (37.5%). DNA of Borrelia spp. was identified in 37.5% of ticks. Thus, the combined method of mechanical homogenization of ticks in combination with a commercial kit for DNA isolation, offers maximum efficiency in terms of speed, number and size of samples to be studied.

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An Alternative and Rapid Method for the Extraction of Nucleic Acids from Ixodid Ticks by Potassium Acetate Procedure

Abstract: ABSTRACT

Cited by 17 publications

References 22 publications

Diversity of Rickettsia species in collected ticks from Southeast Iran

Diversity of Rickettsia species in collected ticks from Southeast Iran

First Report on Truffle-Inhabiting Fungi and Metagenomic Communities of Tuber Aestivum Collected in Russia

Comparison of methods of DNA extraction from ixodid ticks

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