Evaluation of Propidium Monoazide Real-Time PCR for Viablity of Mycobacterium leprae / 나학회지
Korean Leprosy Bulletin
;
: 13-22, 2016.
Artigo
em Coreano
| WPRIM
| ID: wpr-222629
ABSTRACT
BACKGROUND:
Conventional acid-fast bacilli (AFB) staining cannot differentiate viable from dead cells. Propidium monoazide (PMA) is a photoreactive DNA-binding dye that inhibits PCR amplification by DNA modification. OBJECT The author evaluated whether PMA real-time PCR is suitable for the viablity of Mycobacterium leprae (M. leprae) in specimens of cultivation in mouse foot pads.METHODS:
A total of 55 diluted suspensions from mouse foot pads were quadruplicated and subjected to PMA treatment and/or heat inactivation, and were also tested to compare the ΔCT values (CT value in PMA-treated samples-CT value in non-PMA-treated samples). Real-time PCR was performed using QuantiTect SYBR® Green PCR Kits(Qiagen, USA), and the CT value changes after PMA treatment were compared between PMA treatment and/or heat inactivation groups.RESULTS:
The increase in the CT value after PMA treatment was significant in heat inactivated group(4.26) and non-heat inactivated group(1.12)(both P = 0.000). In the ROC curve analysis, the cutoff ΔCT value for maximum sensitivity (100%) and specificity (97.1%) for differentiating dead from live cells was 2.41CONCLUSIONS:
PMA real-time PCR is a useful approach for evaluating viablity of M. leprae.
Texto completo:
DisponíveL
Índice:
WPRIM (Pacífico Ocidental)
Assunto principal:
Propídio
/
Suspensões
/
DNA
/
Reação em Cadeia da Polimerase
/
Curva ROC
/
Sensibilidade e Especificidade
/
Reação em Cadeia da Polimerase em Tempo Real
/
Pé
/
Temperatura Alta
/
Mycobacterium
Tipo de estudo:
Estudo diagnóstico
/
Estudo prognóstico
Limite:
Animais
Idioma:
Coreano
Revista:
Korean Leprosy Bulletin
Ano de publicação:
2016
Tipo de documento:
Artigo
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