Rapid point-of-care diagnostics for the detection of Chlamydia pecorum in koalas (Phascolarctos cinereus) using loop-mediated isothermal amplification without nucleic acid purification.
Microbiologyopen
; 8(12): e916, 2019 12.
Article
em En
| MEDLINE
| ID: mdl-31419041
ABSTRACT
Infectious disease, predominately chlamydiosis, contributes significantly to the decline in health of wild koala (Phascolarctos cinereus) populations in some regions of Australia. In this study, we describe the development and evaluation of a simple, sensitive, and specific loop-mediated isothermal amplification (LAMP) assay for the detection of Chlamydia pecorum in koalas as a point-of-care diagnostic tool that can be used in any wildlife hospital and in the field on specialized instrumentation. A set of primers targeting a 188-bp region of the C. pecorum genome was designed. 100% specificity of the LAMP assay was revealed by demonstrating no cross-reactivity with 33 nontarget pathogens, and complete correlation with qPCR results for 43 clinical swabs collected opportunistically from wildlife hospitals. In sensitivity evaluations, the technique successfully detected serial dilutions of extracted C. pecorum DNA with a detection limit of 44 IFU/ml.
Palavras-chave
Texto completo:
1
Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Infecções por Chlamydia
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Chlamydia
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Técnicas de Amplificação de Ácido Nucleico
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Phascolarctidae
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Testes Imediatos
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Doenças dos Animais
Tipo de estudo:
Diagnostic_studies
Limite:
Animals
Idioma:
En
Revista:
Microbiologyopen
Ano de publicação:
2019
Tipo de documento:
Article