Rapid detection of mecA and spa by the loop-mediated isothermal amplification (LAMP) method.
OBJECTIVE
To develop a detection assay for staphylococcal mecA and spa by using loop-mediated isothermal amplification (LAMP) method.


RESULTS
Staphylococcus aureus and other related species were subjected to the detection of mecA and spa by both PCR and LAMP methods. The LAMP successfully amplified the genes under isothermal conditions at 64 degrees C within 60 min, and demonstrated identical results with the conventional PCR methods. The detection limits of the LAMP for mecA and spa, by gel electrophoresis, were 10(2) and 10 cells per tube, respectively. The naked-eye inspections were possible with 10(3) and 10 cells for detection of mecA and spa, respectively. The LAMP method was then applied to sputum and dental plaque samples. The LAMP and PCR demonstrated identical results for the plaque samples, although frequency in detection of mecA and spa by the LAMP was relatively lower for the sputum samples when compared to the PCR methods.


CONCLUSIONS
Application of the LAMP enabled a rapid detection assay for mecA and spa. The assay may be applicable to clinical plaque samples.


CONCLUSIONS
The LAMP offers an alternative detection assay for mecA and spa with a great advantage of the rapidity.
Department of Pathophysiology - Periodontal Science, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama, Japan.
Y
Y
Koide
H
H
Maeda
K
K
Yamabe
K
K
Naruishi
T
T
Yamamoto
S
S
Kokeguchi
S
S
Takashiba
D000368:Aged; D000369:Aged, 80 and over; D001426:Bacterial Proteins / Q000032:analysis*; D001431:Bacteriological Techniques; D004269:DNA, Bacterial / Q000302:isolation & purification; D003773:Dental Plaque / Q000382:microbiology; D006801:Humans; D057230:Limit of Detection; D055624:Methicillin-Resistant Staphylococcus aureus / Q000235:genetics / Q000302:isolation & purification*; D008875:Middle Aged; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D046915:Penicillin-Binding Proteins; D016133:Polymerase Chain Reaction; D012680:Sensitivity and Specificity; D013205:Staphylococcal Protein A / Q000032:analysis*
D001426:Bacterial Proteins; D004269:DNA, Bacterial; D046915:Penicillin-Binding Proteins; D013205:Staphylococcal Protein A; C487789:mecA protein, Staphylococcus aureus