等位基因特异性PCR法快速检测烟曲霉常见耐药突变基因

中国真菌学杂志　 2021, Vol. 16 　Issue (6): 388-391.

论著上一篇

等位基因特异性PCR法快速检测烟曲霉常见耐药突变基因

邓宇晨¹, 方文捷¹, 邓淑文¹, 陈显振¹, 张亚茹², 冯真益², 凌丽燕², 潘炜华¹, 廖万清¹

1. 海军军医大学长征医院皮肤科, 上海 200003;
2. 浙江省平湖市第二人民医院检验科, 平湖 314201

收稿日期:2021-06-09 发布日期:2021-12-29
通讯作者: 潘炜华,E-mail:panweihua9@sina.com;凌丽燕,E-mail:1144623706@qq.com E-mail:panweihua9@sina.com;1144623706@qq.com
作者简介:邓宇晨,男(汉族),博士研究生在读. E-mail: smmudeng@outlook.com
基金资助:
嘉兴市科技计划项目（2019AY32015）；第二军医大学精准医学转化应用研究专项（2017JZ49）

Allele specific PCR for rapid detection of common gene mutations associated with the resistance of Aspergillus fumigatus

DENG Yuchen¹, FANG Wenjie¹, DENG Shuwen¹, CHEN Xianzhen¹, ZHANG Yaru², FENG Zhenyi², LING Liyan², PAN Weihua¹, LIAO Wanqing¹

1. Department of Dermatology, Changzheng Hospital, Naval Military Medical University, Shanghai 200003, China;
2. Department of Clinical Laboratory, The No.2 People's Hospital of Pinghu, Pinghu 314201, China

Received:2021-06-09 Published:2021-12-29

摘要/Abstract

摘要： 目的建立一套检测烟曲霉L98H/TR34耐药突变的二重扩增体系,进而快速准确判断其对于唑类的耐药情况。方法通过等位基因特异性PCR法实现L98H/TR34耐药突变二重PCR检测,评价该体系的特异性和敏感性。结果所有烟曲霉L98H/TR34耐药基因阳性菌株实时荧光定量PCR(Real Time PCR, RT-PCR)溶解曲线均呈现双峰,无突变的菌株均未检测到突变位点。结论等位基因特异性PCR能够快速检测烟曲霉菌常见耐药突变基因L98H/TR34,具有较高的特异性和敏感性,操作简单,价格低廉。

关键词: 烟曲霉, 等位基因特异性扩增, TR34/L98H

Abstract: Objective To establish an allele specific PCR detection system for common resistance mutation genes L98H/TR34 of Aspergillus fumigatus and determine its resistance to azoles quickly and accurately. Methods Allele specific PCR method was used to simultaneously detect azole-resistant mutation genes L98H/TR34, and specificity and sensitivity of the system was evaluated. Results The real time PCR dissolution curves of all resistance gene positive Aspergillus fumigatus showed double peaks, but no mutation sites were detected in non-resistant Aspergillus fumigatus. Conclusion Allele-specific PCR can quickly detect common resistance mutant genes L98H/TR34 of Aspergillus fumigatus, with high specificity and sensitivity, simple operation and low price.

Key words: Aspergillus fumigatus, allele specific PCR, TR34/L98H

中图分类号:

R379.6

邓宇晨, 方文捷, 邓淑文, 陈显振, 张亚茹, 冯真益, 凌丽燕, 潘炜华, 廖万清. 等位基因特异性PCR法快速检测烟曲霉常见耐药突变基因[J]. 中国真菌学杂志, 2021, 16(6): 388-391.

DENG Yuchen, FANG Wenjie, DENG Shuwen, CHEN Xianzhen, ZHANG Yaru, FENG Zhenyi, LING Liyan, PAN Weihua, LIAO Wanqing. Allele specific PCR for rapid detection of common gene mutations associated with the resistance of Aspergillus fumigatus[J]. Chinese Journal of Mycology, 2021, 16(6): 388-391.

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等位基因特异性PCR法快速检测烟曲霉常见耐药突变基因

Allele specific PCR for rapid detection of common gene mutations associated with the resistance of Aspergillus fumigatus

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