Journal of Pure and Applied MicrobiologyVol. 9 No. Special Edition Nov. 2015

Clinical Application of Loop-Mediated Isothermal Amplification in the Rapid Detection of Methicillin-Resistant Staphylococcus aureus

Cai-Yan Li*#, Wen-Feng Huang#, E Cai and Qun-Li Wang

Clinical Laboratory Medicine Center, The Second People?s Hospital of Jingmen, China.

Received on 02 June 2015 and accepted on 10 August 2015

 

ABSTRACT

To develop a detection assay for staphylococcal mecA and spa by using loop-mediated isothermal amplification (LAMP) method. Staphylococcus aureus and other related species were subjected to the detection of mecAand spa by both PCR and LAMP methods. The LAMP successfully amplified the genes under isothermal conditions at 64! within 60 min, and demonstrated identical results with the conventional PCR methods. The detection limits of the LAMP for mecA and spa, by gel electro-phoresis, were 102 and 10 cells per tube, respectively. The naked-eye inspec-tions were possible with 103 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. Application of the LAMP enabled a rapid detection assay for mecA and spa. The assay may be applicable to clinical plaque samples. The LAMP offers an alternative detection assay for mecA and spa with a great advantage of the rapidity.

Keywords : loop-mediated isothermal amplification LAMP, Methicillin-resistant Staphylococcus aureus, mecA.