Optimization of PMA-qPCR for Staphylococcus aureus and determination of viable bacteria in indoor air.
Indoor Air
; 28(1): 64-72, 2018 Jan.
Article
em En
| MEDLINE
| ID: mdl-28683164
ABSTRACT
Staphylococcus aureus may cause infections in humans from mild skin disorders to lethal pneumonia. Rapid and accurate monitoring of viable S. aureus is essential to characterize human exposure. This study evaluated quantitative PCR (qPCR) with propidium monoazide (PMA) to quantify S. aureus. The results showed comparable S. aureus counts between exclusively live cells and mixtures of live/dead cells by qPCR with 1.5 or 2.3 µg/mL PMA (P>.05), illustrating the ability of PMA-qPCR to detect DNA exclusively from viable cells. Moreover, qPCR with 1.5 or 2.3 µg/mL PMA performed optimally with linearity over 103 -108 CFU/mL (R2 ≥0.9), whereas qPCR with 10, 23 or 46 µg/mL PMA significantly underestimated viable counts. Staphylococcus aureus and total viable bacteria were further determined with PMA-qPCR (1.5 µg/mL) from 48 samples from a public library and two university dormitories and four from outside. Viable bacteria averaged 1.9×104 cells/m3 , and S. aureus were detected in 22 (42%) samples with a mean of 4.4×103 cells/m3 . The number of S. aureus and viable bacteria were positively correlated (r=.61, P<.005), and percentages of S. aureus relative to viable bacteria averaged 12-44%. The results of field samples suggest that PMA-qPCR can be used to quantify viable S. aureus cells.
Palavras-chave
Texto completo:
1
Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Staphylococcus aureus
/
Microbiologia do Ar
/
Carga Bacteriana
Tipo de estudo:
Evaluation_studies
Idioma:
En
Revista:
Indoor Air
Assunto da revista:
SAUDE AMBIENTAL
Ano de publicação:
2018
Tipo de documento:
Article
País de afiliação:
Taiwan