Improvement on DNA extraction method of methicillin resistant Staphylococcus aureus / 国际检验医学杂志

ABSTRACT

Objective To study a nucleic acid extraction method suitable for detecting methicillin‐resistant Staphylococcus aureus (MRSA) by PCR method .Methods Under different incubation conditions ,MRSA was cracked by lysozyme ,lysostaphin or chel‐ex100R resin for obtaining DNA ,then the target gene was detected by using the PCR method .Results DNA was obtained by sim‐ultaneously using lysozyme ,lysostaphin and chelex100R resin solution ,the obtained Ct value was significantly lower than that of the other components of schizolysis solutions when PCR was used to detect mecA gene of obtained DNA .There was no statistically sig‐nificant difference between adopting the 56 ℃ one‐step method and the 37 ℃ and 56 ℃ two‐step method for conducting MRSA schizolysis(P> 0 .05) ,but the steps were simplified .Conclusion Incubating MRSA in solution containing lysozyme ,lysostaphin , chelex100R resin for 30 min at 56 ℃ is the convenient and efficient schizolysis method to extract DNA ,which can be used immedi‐ately for the next step of PCR and lays the foundation for PCR rapid detection of clinical MRSA infection .