Abstract
In Africa, pathogens transmitted by ticks are of major concern in livestock production and human health. Despite noticeable improvements particularly of molecular screening methods, their widespread availability and the detection of multiple infections remain challenging. Hence, we developed a universally accessible and robust tool for the detection of bacterial pathogens and piroplasmid parasites of cattle. A low-cost and low-density chip DNA microarray kit (LCD-Array) was designed and tested towards its specificity and sensitivity for five genera causing tick-borne diseases. The blood samples used for this study were collected from cattle in Northern Cameroon. Altogether, 12 species of the genera Anaplasma, Ehrlichia, Rickettsia and Theileria, and their corresponding genus-wide probes including Babesia were tested on a single LCD-Array. The detection limit of plasmid controls by PCR ranged from 1 to 75 copies per µL depending on the species. All sequenced species hybridized on the LCD-Array. As expected, PCR, agarose gel electrophoresis and Sanger sequencing found significantly less pathogens than the LCD-Array (p < 0.001). Theileria and Rickettsia had lower detection limits than Anaplasma and Ehrlichia. The parallel identification of some of the most detrimental tick-borne pathogens of livestock, and the possible implementation in small molecular-diagnostic laboratories with limited capacities makes the LCD-Array an appealing asset.
Highlights
Tick-borne pathogens (TBP) are of high veterinary and medical importance worldwide
For cattle and other livestock, the published literature highlights the importance of protozoa of the genera Babesia and Theileria, bacteria of the genera Anaplasma, Ehrlichia and Rickettsia, and arboviruses as etiologic agents of many diseases, of which a number of them have zoonotic potential [1]
In developing countries, routine diagnostic approaches for the identification of TBPs are generally based on microscopic examination of blood smears [2,3] or serological assays [4,5]
Summary
Tick-borne pathogens (TBP) are of high veterinary and medical importance worldwide. In developing countries, routine diagnostic approaches for the identification of TBPs are generally based on microscopic examination of blood smears [2,3] or serological assays [4,5]. For the identification of TBPs are generally based on microscopic examination of blood smears [2,3] orTsroepr.oMloegd.icInaflecat.sDsaisy. N not financially affordable for veterinary laboratories in low income endemic couTnhterieDs.NMAolmeciucrloarartroaoylstbeachsendoloongyPCoRf [P1C0]Ra-namd pnloiwfieaddapyrsoNduGcStsarceobmebcionmesinhgigmhortehwroiudgehspprueta, d, sewnsiitthivNitGy,Sspbeeicnifgiceitcyonaonmd irceaplrlyodvuiacbibleilwityhe[1n2u].sIetds ffuonr clatirogne issabmapseledsoiznetsh[e11re].verse line blot (RLB), in which sTpheecDifiNc Aolmigiocnrouacrlreaoytitdeechsnpooltosg(yporof bPeCsR) -aarme pimlifimeodbpilriozdeudcotsncaomsobliindessuhrifgahceth(rFoiugguhrepu1)t,. Ized for minimal input of equipment, workflow, costs and expenditure of time, and suitable for small diagnostic laboratories in low and middle income developing countries [14,15] AItssefuqnucetniocenisisabdadseedd, iotnhythberidreivzeesrswe iltihnethbeloptro(RbeLBw)h, einrewithisich destpecetceidficboyliagoflnuuocrleesocteidnte, scphoetms i(lpurmobineess)caernetimormboiobtiilnizyeldatoedn alasboelli.dTshuerfsaycnec(hFriognuoreus1)d. eWtehcteinona otafraget musaltmitupdlee wofithspceocmiesplienmtehnetasraymDe NgeAnesteicqumenatceeriiasl ahdads ecdo,nittrihbyubterdidtizoesitswpitohputhlaeriptyroibne iwnfhecetrieouits is disdeeatseectdediagbnyoastflicuso[r1e0s,c1e3n].t,Lcohwem-dielunsmitiyneDsNceAntmoricbroioatrirnayylsatseudchlabaselt.hTehLeCsDyn-Achrrraoynoaures ddeesteigcntieodn toof a demteuctltmituudceholof wspeercnieusminbtehres osafmpaetgheongeetnicicmaagteenrtiaslthhaasnchoingthr-idbuentesditytomitiscrpooapruralayrsit[y14in]. iHnfoewcteiovuesr,dthiseeyase arediaogpntiomstizicesd[1f0o,r13m].inLiomwa-ldiennpsuittyoDf NeqAumipimcreonatr,rawyosrskuflcohwa,sctohsetsLCanDd-Aerxrpaeynadrietudreesiogfnteidmteo, danetdect themreufcohreloswuietrabnluemfobresrms oalflpdaitahgongoesntiicc laagbeonratstotrhiaens ihniglohw-deannsditmy imddiclreoianrcroamyse[1d4e]v.eHloopwinegvceor,utnhteryieasre [14o,p15ti]m. ized for minimal input of equipment, workflow, costs and expenditure of time, and suitable for small diagnostic laboratories in low and middle income developing countries [14,15]
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