Prevalence and extent of heteroresistance by next generation sequencing of multidrug-resistant tuberculosis.
Prevalence and extent of heteroresistance by next generation sequencing of multidrug-resistant tuberculosis.
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DOI：
10.1371/journal.pone.0176522
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发表时间：
2017
期刊：
影响因子：
3.7
通讯作者：
Houpt ER
中科院分区：
文献类型：
--
作者：
Operario DJ;Koeppel AF;Turner SD;Bao Y;Pholwat S;Banu S;Foongladda S;Mpagama S;Gratz J;Ogarkov O;Zhadova S;Heysell SK;Houpt ER
基于扩增子的下一代测序(NGS)是一种新兴的结核分枝杆菌药敏试验方法，但尚未得到很好的描述。我们通过覆盖3519个核苷酸的11个耐药相关基因区域的NGS检测了158株临床多药耐药结核分枝杆菌。在这些基因区域中，6.3%的基因座中至少有一个菌株存在完全抗性或异质性抗性(定义为1%-99%的突变)。异抗菌株的数量以gyrA密码子94、rpoB密码子526和531、embB密码子306、372和406最高(11-26%的菌株表现出异抗性)。57%的MDR菌株对一个或多个已知的耐药相关突变具有异质性耐药。异源抗性基因座通常表现出高或低程度的突变(>；90%或<；10%)。NGS对检测低水平PncA异质性抗性的高度敏感性似乎改善了PZA的基因-表型敏感性相关性。NGS表明，结核病中关键基因区域的异质性耐药是常见的，需要进行生物信息管理。这种异质性耐药的临床意义尚不清楚，应继续对pNcA进行进一步的研究。
Amplicon-based Next Generation Sequencing (NGS) is an emerging method for Mycobacterium tuberculosis drug susceptibility testing (DST) but has not been well described. We examined 158 clinical multidrug-resistant M. tuberculosis isolates via NGS of 11 resistance-associated gene regions covering 3519 nucleotides. Across these gene regions, complete resistance or heteroresistance (defined as 1%-99% mutation) was present in at least one isolate in 6.3% of loci. The number of isolates with heteroresistance was highest for gyrA codon 94, rpoB codons 526 and 531, and embB codons 306, 372 and 406 (range 11–26% of isolates exhibited heteroresistance). 57% of MDR strains had heteroresistance of one or more recognized resistance-associated mutation. Heteroresistant loci generally exhibited high or low degrees of mutation (>90% or <10%). The deep sensitivity of NGS for detecting low level pncA heteroresistance appeared to improve genotypic-phenotypic PZA susceptibility correlations over that of Sanger. NGS demonstrates that heteroresistance in TB in the regions of key genes is common and will need to be bioinformatically managed. The clinical significance of such heteroresistance is unclear, and further study of pncA should be pursued.