Molecular detection of mecA gene from Methicillin Resistant Coagulase Negative Staphylococci

Arun Thangave, C. Rajamanickam, O. Sahu, S. Ponnappan, Gezahegn Abawa, Abenezer Tadele
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引用次数: 2

Abstract

In the present research, a total of 27 Wound swab samples were collected for the isolation of Methicillin Resistant-Coagulase Negative Staphylococci. Out of these, 3.7% of the samples have been shown the presence of Methicillin Resistant-CoagulaseNegative Staphylococci (MR-CoNS). The preliminary coagulase activity has been identified by Coagulase slide and tube method. The HiCrome MeReSa agar medium has been used for the preliminary identification of Methicillin resistance. During the isolation, the other strains of Coagulase Negative Staphylococci (29.6%), Methicillin Sensitive Staphylococcus aureus (44.4%) have been also isolated. 6 samples (22.2%) were sterile. The genomic DNA was isolated from Methicillin resistant Coagulase Negative Staphylococciand other species of Staphylococcithrough Modified DNA extraction method. The isolated DNA was amplified through the thermal cycler using suitable forwardand reverse oligonucleotide primers, for the amplification of 527bp of mecA gene. The amplified DNA product has been identified by agarose gel electrophoresis using suitable 100bp ladder marker.
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耐甲氧西林凝固酶阴性葡萄球菌mecA基因的分子检测
本研究收集了27份伤口拭子样本进行甲氧西林耐药凝固酶阴性葡萄球菌的分离。其中,3.7%的样本显示存在耐甲氧西林凝血阴性葡萄球菌(MR-CoNS)。用凝固酶玻片法和试管法初步测定了凝固酶活性。采用hrome MeReSa琼脂培养基对甲氧西林耐药进行初步鉴定。在分离过程中,还分离出凝固酶阴性葡萄球菌(29.6%)和甲氧西林敏感金黄色葡萄球菌(44.4%)。无菌6份(22.2%);采用改良DNA提取法从耐甲氧西林凝固酶阴性葡萄球菌和其他葡萄球菌中分离出基因组DNA。采用合适的正、反寡核苷酸引物对分离的DNA进行热循环扩增,扩增出527bp的mecA基因。扩增的DNA产物经琼脂糖凝胶电泳鉴定,采用合适的100bp阶梯标记。
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