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Development and validation of 2 probe-hybridization quantitative PCR assays for rapid detection of a pathogenic Coxiella species in captive psittacines.


ABSTRACT: Avian coxiellosis is an emerging cause of morbidity and mortality among captive psittacines, and the utility of a rapid detection test using easily obtained samples is paramount in a clinical setting. New sequences were obtained from 3 genes: groEL, dnaK, and rpoB. We developed probe-hybridization quantitative PCR (qPCR) assays using groEL and dnaK genes. Samples, including splenic aspirates, liver aspirates, whole blood, and choanal, conjunctival, and cloacal swabs, were collected from 4 psittacine species including 3 blue-and-gold macaws (Ara ararauna), 2 scarlet-chested parrots (Neophema splendida), 1 Timneh African grey parrot (Psittacus timneh), and 1 yellow-naped Amazon parrot (Amazona auropalliata). Retrospective review of postmortem findings from 3 of these psittacines included splenomegaly, hepatitis, and/or transmission electron microscopy confirmation consistent with previous reports of avian coxiellosis. There was 100% agreement between these assays and consensus PCR with sequencing. A Wilcoxon rank-sum test found a strong correlation between groEL and dnaK cycle threshold values (p < 0.001), validating these assays for detection of this avian Coxiella sp.

SUBMITTER: Flanders AJ 

PROVIDER: S-EPMC7377607 | biostudies-literature | 2020 May

REPOSITORIES: biostudies-literature

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Development and validation of 2 probe-hybridization quantitative PCR assays for rapid detection of a pathogenic <i>Coxiella</i> species in captive psittacines.

Flanders Alison J AJ   Speer Brian B   Reavill Drury R DR   Roberts John F JF   Childress April L AL   Atlas Alvin A   Wellehan James F X JFX  

Journal of veterinary diagnostic investigation : official publication of the American Association of Veterinary Laboratory Diagnosticians, Inc 20200319 3


Avian coxiellosis is an emerging cause of morbidity and mortality among captive psittacines, and the utility of a rapid detection test using easily obtained samples is paramount in a clinical setting. New sequences were obtained from 3 genes: <i>groEL, dnaK</i>, and <i>rpoB</i>. We developed probe-hybridization quantitative PCR (qPCR) assays using <i>groEL</i> and <i>dnaK</i> genes. Samples, including splenic aspirates, liver aspirates, whole blood, and choanal, conjunctival, and cloacal swabs,  ...[more]

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