Abstract
The increasing resistance of staphylococci to ß -lactam antibiotics has become a major clinical problem. Development of rapid and sensitive techniques for detection of MRSA is an important aim for public health. A duplex PCR were established for specific identification of methicillin-resistance Staphylococcus aureus (MRSA) in clinical samples. In this work a duplex SYBR Green real time PCR was developed for rapid identification of MRSA in local methicillin-resistance S. aureus isolates. Twenty methicillin-resistance S. aureus isolates, as determined by disc diffusion method, were subjected to DNA extraction and PCR amplification. Two genes were amplified successfully, mecA (533bp) and femA (314bp), as targets for methicillin-resistance and specific identification of S. aureus, respectively using conventional PCR. Sensitivity of the duplex PCR showed that the minimum concentration of DNA that gave positive results for the two genes was 30ng/µl. In order to develop rapid and sensitive test for identification of MRSA, serial dilutions of purified DNA were amplified gradually according to their concentrations using SYBR Green real time PCR. These results indicated that the SYBR Green real time PCR can be used for identification of methicillin-resistance S. aureus (MRSA) in clinical
Highlights
Staphylococcus aureus is one of the most common human pathogens, especially in hospital patients
In order to develop rapid and sensitive test for identification of methicillin-resistance Staphylococcus aureus (MRSA), serial dilutions of purified DNA were amplified gradually according to their concentrations using SYBR Green real time PCR. These results indicated that the SYBR Green real time PCR can be used for identification of methicillin-resistance S. aureus (MRSA) in clinical samples
Multiplex PCR were developed for rapid identification of MRSA through the amplification of mecA and femA genes [3,4,5]
Summary
Staphylococcus aureus is one of the most common human pathogens, especially in hospital patients. This gram positive bacterium is responsible for various infectious diseases, and has a great capacity to develop resistance to antimicrobial agents [1]. Accurate and rapid identification of methicillin-resistant Staphylococcus aureus (MRSA) in clinical specimens is essential for timely decisions about isolation procedures and effective antimicrobial chemotherapy. The femA gene encodes a factor which is essential for methicillin resistance and is universally present in all S. aureus). The aim of this work is to develop SYBR Green real time PCR for identification of methicillin-resistance Staphylococcus aureus (MRSA) isolated from clinical samples in Iraq
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