Hard ticks are important vectors of DNA- and RNA-based infectious microorganisms, but they also host complex microbial communities in which pathogens and symbionts can interact among each other and with the arthropod host itself. Molecular investigations on ticks and their hosted microorganisms are important for human and animal health. These analyses often imply the use of both DNA and RNA, with prompt preservation of nucleic acids after collection, and safe handling in case of low-level containment. Several commercial kits are available for the co-extraction of DNA and RNA; however, cost can be a limiting factor for the choice of this method, which also require additional reagents for nucleic acids preservation before extraction. Additionally, extraction buffers provided in commercial kits do not guarantee the safe handling in case of hazardous biological material. With the aim of epidemiological screenings for tick-borne pathogens and gene expression analyses focused on the relationship among ticks and their microbial communities, an optimized protocol for DNA and RNA co-extraction from single adult hard tick specimens (Ixodidae) has been developed using TRIzolⓇ LS Reagent.

Protocol optimization for simultaneous DNA and RNA co-extraction from single hard tick specimens / A. Cafiso, G. Chiappa, C. Luzzago, A. Koni, D. Bonato, X. Koleci, C. Bazzocchi. - In: METHODSX (AMSTERDAM). - ISSN 2215-0161. - 8(2021 Jan), pp. 101315.1-101315.6. [10.1016/j.mex.2021.101315]

Protocol optimization for simultaneous DNA and RNA co-extraction from single hard tick specimens

A. Cafiso
Co-primo
;
G. Chiappa
Co-primo
;
C. Luzzago
Secondo
;
C. Bazzocchi
Ultimo
2021

Abstract

Hard ticks are important vectors of DNA- and RNA-based infectious microorganisms, but they also host complex microbial communities in which pathogens and symbionts can interact among each other and with the arthropod host itself. Molecular investigations on ticks and their hosted microorganisms are important for human and animal health. These analyses often imply the use of both DNA and RNA, with prompt preservation of nucleic acids after collection, and safe handling in case of low-level containment. Several commercial kits are available for the co-extraction of DNA and RNA; however, cost can be a limiting factor for the choice of this method, which also require additional reagents for nucleic acids preservation before extraction. Additionally, extraction buffers provided in commercial kits do not guarantee the safe handling in case of hazardous biological material. With the aim of epidemiological screenings for tick-borne pathogens and gene expression analyses focused on the relationship among ticks and their microbial communities, an optimized protocol for DNA and RNA co-extraction from single adult hard tick specimens (Ixodidae) has been developed using TRIzolⓇ LS Reagent.
Nucleic acids; Purification; DNA isolation; RNA isolation; Co-extraction; Tick; PCR; Reverse transcription; Coinfection; Vector-borne pathogen
Settore VET/06 - Parassitologia e Malattie Parassitarie degli Animali
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/2434/830202
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