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首页> 外文期刊>International journal of urology: official journal of the Japanese Urological Association >Rapid detection of the fluoroquinolone resistance-associated ParC mutation in Neisseria gonorrhoeae using TaqMan probes.
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Rapid detection of the fluoroquinolone resistance-associated ParC mutation in Neisseria gonorrhoeae using TaqMan probes.

机译：使用TaqMan探针快速检测淋病奈瑟氏球菌中与氟喹诺酮耐药性相关的ParC突变。

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BACKGROUNDS: In fluoroquinolone-resistant Neisseria gonorrhoeae, the amino acid mutations in the fluoroquinolone-resistant determining region (QRDR) of the parC gene are an important factor. The aim of the present study was to develop a rapid detection method of a serine 88 to proline substitution in parC which we previously showed as having significantly higher fluoroquinolone minimal inhibitory concentrations (MIC) using the TaqMan discrimination system. METHODS: We extracted DNA from 90 urine or urethral swab samples obtained from male patients with urethritis caused by N. gonorrhoeae. After DNA extraction, they were subjected to real-time polymerase chain reaction (PCR) using a TaqMan discrimination system and compared with the results of conventional DNA sequencing. RESULTS: Of the 90 samples, the TaqMan technique result showed 13 samples that were classified as having a serine 88 to proline mutation in parC, and 77 samples that did not have a serine 88 to proline mutation in parC. The classifications of all samples completely corresponded to those determined by conventional DNA sequencing. We also found that N. gonorrhoeae with a serine 88 to proline mutation in parC have a significantly higher MIC to ciprofloxacin than that without a serine 88 to proline mutant in parC. CONCLUSIONS: The present genotyping method of real-time PCR using a TaqMan discrimination system could be applied to the rapid detection of a serine 88 to proline amino acid mutation in parC of N. gonorrhoeae. This point mutation is significant for the determination of fluoruquinolone resistance. This rapid detection system may lead to the prevention of use of noneffective antimicrobial agents and a decrease of resistant strains.
机译：背景：在耐氟喹诺酮淋病奈瑟氏球菌中，parC基因的耐氟喹诺酮决定区（QRDR）中的氨基酸突变是重要因素。本研究的目的是开发一种快速检测parC中脯氨酸取代的丝氨酸88的方法，我们先前证明使用TaqMan判别系统具有明显更高的氟喹诺酮最小抑菌浓度（MIC）。方法：我们从90例男性淋病奈瑟菌性尿道炎患者尿液或尿道拭子样本中提取DNA。 DNA提取后，使用TaqMan判别系统对它们进行实时聚合酶链反应（PCR），并与常规DNA测序结果进行比较。结果：在90个样本中，TaqMan技术结果显示13个样本在parC中被分类为具有丝氨酸88脯氨酸突变，而77个样本在parC中没有丝氨酸88发生脯氨酸突变。所有样品的分类完全对应于常规DNA测序确定的分类。我们还发现，在parC中具有丝氨酸88脯氨酸突变的淋病奈瑟氏球菌与在proC中没有丝氨酸88脯氨酸突变的MIC相比具有明显更高的MIC。结论：目前使用TaqMan判别系统进行实时PCR的基因分型方法可用于快速检测淋病奈瑟菌parC中脯氨酸氨基酸突变的丝氨酸88。该点突变对于确定氟喹诺酮耐药性很重要。这种快速检测系统可能导致防止使用无效的抗菌剂并降低耐药菌株。

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