Development and evaluation of genomics informed real-time PCR assays for the detection and strain typing of Mycobacterium avium subsp. paratuberculosis.
J Appl Microbiol
; 135(5)2024 May 01.
Article
em En
| MEDLINE
| ID: mdl-38684472
ABSTRACT
AIMS:
This study aimed to identify specific genomic targets for the detection and strain typing of Map and analyse their sensitivity and specificity, and detect Map directly from faeces. METHODS ANDRESULTS:
A comparative genomics approach was used to identify specific genomic targets for the detection and strain typing of Map. A Map specific qPCR using the primer pair 7132 that targets a DNA segregation ATPase protein was able to detect all strains of Map and is more sensitive than the current Johne's disease PCR assays with a sensitivity of 0.0002 fg µl-1. A strain specific qPCR using the Atsa primer pair that targets the arylsulfase gene was able to differentiate between Type S and Type C strains of Map and was more sensitive than the IS1311 PCR and REA with a sensitivity of 40 fg µl-1 and was specific for Type S Map. Both assays successfully detected Map directly from faeces.CONCLUSION:
This study developed and validated two genomics informed qPCR assays, 7132B Map and Atsa Type S and found both assays to be highly specific and sensitive for the detection of Map from culture and directly from faeces. This is the first time that a probe-based qPCR has been designed and developed for Map strain typing, which will greatly improve the response time during outbreak investigations.Palavras-chave
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Base de dados:
MEDLINE
Assunto principal:
Paratuberculose
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Sensibilidade e Especificidade
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Mycobacterium avium subsp. paratuberculosis
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Genômica
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Fezes
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Reação em Cadeia da Polimerase em Tempo Real
Limite:
Animals
Idioma:
En
Ano de publicação:
2024
Tipo de documento:
Article