Recombinase polymerase amplification (RPA) with lateral flow detection for three Anaplasma species of importance to livestock health

Abstract Anaplasma marginale, A. ovis, and A. phagocytophilum are the causative agents of bovine anaplasmosis, ovine anaplasmosis, and granulocytic anaplasmosis, respectively. The gold standard for diagnosis of post-acute and long-term persistent infections is the serological cELISA, which does not...

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Autores principales:	Andrea Salazar, Francisco M. Ochoa-Corona, Justin L. Talley, Bruce H. Noden
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/c03d28a15f7a49849febd8a942a82f11
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spelling	oai:doaj.org-article:c03d28a15f7a49849febd8a942a82f112021-12-02T16:35:27ZRecombinase polymerase amplification (RPA) with lateral flow detection for three Anaplasma species of importance to livestock health10.1038/s41598-021-95402-y2045-2322https://doaj.org/article/c03d28a15f7a49849febd8a942a82f112021-08-01T00:00:00Zhttps://doi.org/10.1038/s41598-021-95402-yhttps://doaj.org/toc/2045-2322Abstract Anaplasma marginale, A. ovis, and A. phagocytophilum are the causative agents of bovine anaplasmosis, ovine anaplasmosis, and granulocytic anaplasmosis, respectively. The gold standard for diagnosis of post-acute and long-term persistent infections is the serological cELISA, which does not discriminate between Anaplasma species and requires highly equipped laboratories and trained personnel. This study addresses the development of a rapid, isothermal, sensitive, species-specific RPA assays to detect three Anaplasma species in blood and cELISA A. marginale-positive serum samples. Three RPA primer and probe sets were designed targeting msp4 genes of each Anaplasma species and the internal control (GAPDH gene) for each assay. The limit of detection of gel-based or RPA-basic assays is 8.99 × 104 copies/µl = A. marginale, 5.04 × 106 copies/µl = A. ovis, and 4.58 × 103 copies/µl = A. phagocytophilum, and for each multiplex lateral flow or RPA-nfo assays is 8.99 × 103 copies/µl of A. marginale, 5.04 × 103 copies/µl of A. ovis, 4.58 × 103 copies/µl of A. phagocytophilum, and 5.51 × 103 copies/µl of internal control (GAPDH). Although none of the 80 blood samples collected from Oklahoma cattle were positive, the RPA-nfo assays detected all A. marginale cattle blood samples with varying prevalence rates of infection, 83% of the 24 cELISA A. marginale-positive serum samples, and all A. phagocytophilum cell culture samples. Overall, although early detection of three Anaplasma species was not specifically addressed, the described RPA technique represents an improvement for detection of three Anaplasma in regions where access to laboratory equipment is limited.Andrea SalazarFrancisco M. Ochoa-CoronaJustin L. TalleyBruce H. NodenNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 11, Iss 1, Pp 1-13 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Andrea Salazar Francisco M. Ochoa-Corona Justin L. Talley Bruce H. Noden Recombinase polymerase amplification (RPA) with lateral flow detection for three Anaplasma species of importance to livestock health
description	Abstract Anaplasma marginale, A. ovis, and A. phagocytophilum are the causative agents of bovine anaplasmosis, ovine anaplasmosis, and granulocytic anaplasmosis, respectively. The gold standard for diagnosis of post-acute and long-term persistent infections is the serological cELISA, which does not discriminate between Anaplasma species and requires highly equipped laboratories and trained personnel. This study addresses the development of a rapid, isothermal, sensitive, species-specific RPA assays to detect three Anaplasma species in blood and cELISA A. marginale-positive serum samples. Three RPA primer and probe sets were designed targeting msp4 genes of each Anaplasma species and the internal control (GAPDH gene) for each assay. The limit of detection of gel-based or RPA-basic assays is 8.99 × 104 copies/µl = A. marginale, 5.04 × 106 copies/µl = A. ovis, and 4.58 × 103 copies/µl = A. phagocytophilum, and for each multiplex lateral flow or RPA-nfo assays is 8.99 × 103 copies/µl of A. marginale, 5.04 × 103 copies/µl of A. ovis, 4.58 × 103 copies/µl of A. phagocytophilum, and 5.51 × 103 copies/µl of internal control (GAPDH). Although none of the 80 blood samples collected from Oklahoma cattle were positive, the RPA-nfo assays detected all A. marginale cattle blood samples with varying prevalence rates of infection, 83% of the 24 cELISA A. marginale-positive serum samples, and all A. phagocytophilum cell culture samples. Overall, although early detection of three Anaplasma species was not specifically addressed, the described RPA technique represents an improvement for detection of three Anaplasma in regions where access to laboratory equipment is limited.
format	article
author	Andrea Salazar Francisco M. Ochoa-Corona Justin L. Talley Bruce H. Noden
author_facet	Andrea Salazar Francisco M. Ochoa-Corona Justin L. Talley Bruce H. Noden
author_sort	Andrea Salazar
title	Recombinase polymerase amplification (RPA) with lateral flow detection for three Anaplasma species of importance to livestock health
title_short	Recombinase polymerase amplification (RPA) with lateral flow detection for three Anaplasma species of importance to livestock health
title_full	Recombinase polymerase amplification (RPA) with lateral flow detection for three Anaplasma species of importance to livestock health
title_fullStr	Recombinase polymerase amplification (RPA) with lateral flow detection for three Anaplasma species of importance to livestock health
title_full_unstemmed	Recombinase polymerase amplification (RPA) with lateral flow detection for three Anaplasma species of importance to livestock health
title_sort	recombinase polymerase amplification (rpa) with lateral flow detection for three anaplasma species of importance to livestock health
publisher	Nature Portfolio
publishDate	2021
url	https://doaj.org/article/c03d28a15f7a49849febd8a942a82f11
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Recombinase polymerase amplification (RPA) with lateral flow detection for three Anaplasma species of importance to livestock health

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