A new simple assay for determining aminoglycoside inactivation in intact cells of Pseudomonas aeruginosa.
A new simple assay for determining aminoglycoside inactivation in intact cells of Pseudomonas aeruginosa.
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一种用于确定铜绿假单胞菌完整细胞中氨基糖苷失活的新的简单测定法。
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发表时间：
1997
期刊：
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通讯作者：
T. Sawai
中科院分区：
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作者：
K. O'hara;T. Kawabe;K. Taniguchi;M. Ohnuma;M. Nakagawa;Y. Naitou;T. Sawai
氨基糖苷(AG)耐药铜绿假单胞菌的完整细胞通常不会灭活AG，即使它们拥有AG修饰酶。先前报道了一种测定耐链霉素铜绿假单胞菌完整细胞失活酶活性的方法。该方法虽然适用于卡那霉素(Km)失活酶的测定，但不适用于部分Km抗性菌株。现已研究了一种新的测定临床分离耐Km铜绿假单胞菌完整细胞Km失活酶活性的简便方法。尝试用溶菌酶在洗涤细胞中释放失活酶，并同时加入DNA酶和核糖核酸酶来消化细菌分解释放的核酸，以测定AG失活酶的活性。这种溶菌酶-DNA酶-核糖核酸酶(LDR)方法有助于确认所用菌株中是否存在AG失活酶。此外，LDR技术也适用于除KM以外的各种AGG的失活酶活力的测定。由于这种简单的测定方法可以测定不同类型铜绿假单胞菌的任何类型的AG失活酶活性，因此它可能对临床上快速选择药物有重要意义。
Intact cells of aminoglycoside (AG) antibiotic-resistant Pseudomonas aeruginosa usually do not inactivate AG, even though they possess the AG-modifying enzyme. An assay method for determining the activity of inactivating enzyme in intact cells of streptomycin-resistant P. aeruginosa was previously reported. Although this assay method was applied to the determination of the activity of kanamycin (KM)-inactivating enzymes, it could not apply to some of the KM-resistant strains. A new simple assay method has now been investigated for determining the activity of KM-inactivating enzyme in intact cells of clinically isolated KM-resistant P. aeruginosa. The determination of AG-inactivating enzyme activity was attempted using lysozyme for release of the inactivating enzymes in washed cells, and both DNase and RNase were added to digestion of the nucleic acids released by bacteriolysis. This lysozyme-DNase-RNase (LDR) method has facilitated the confirmation of the presence of AG-inactivating enzyme in the strains used. In addition, the LDR technique was applicable to the determination of inactivating enzyme activity for various AGs other than KM. Since this simple assay method can determine any type of AG-inactivating enzyme activity of various P. aeruginosa strains, it may contribute significantly to the rapid selection of drugs in clinical use.