高通量测序技术在慢性咽炎患者咽部细菌耐药基因检测中的应用价值

摘要

目的：探讨高通量测序技术在慢性咽炎患者耐药基因分布中的应用。方法选取2010年5月至2012年5月本院门诊收治的慢性咽炎患者360例，采集患者咽后壁分泌物，提取质粒 DNA 并进行 Solexa 高通量测序，通过比对在 NCBI 质粒基因组数据库进行物种鉴定，通过与 ARDB 数据库进行比对进行耐药基因鉴定。结果本研究共鉴定出307个质粒，分属25种病原菌，所占比例最高的为α-溶血性链球菌（12．87％），其次为副流感嗜血菌（9．86％）。通过与 ARDB 中已知的380种 ARG 比对，共有4015条序列与已知耐药基因序列一致。其中氨基糖苷类耐药基因、β内酰胺酶类耐药基因、大环内酯类耐药基因及四环素类耐药基因占较高的比例。同时我们也检测到一个多重耐药基因 lsa。结论高通量测序技术可以高效、准确地检测慢性咽炎患者中咽部细菌的耐药基因，为临床慢性咽炎治疗方案的选择提供理论基础。%Objective To explore the application of high-throughput sequencing technology in resistance gene detec-tion in patients with chronic pharyngitis.Methods 360 cases of patients with chronic pharyngitis were selected from May 2010 to May 2012 in outpatient of our hospital.Secretion of pharyngeal wall in patients with chronic pharyngitis was collected and the plasmid was extracted for further Solexa high-throughput sequencing.Species identification was performed through mapping to NCBI plasmid genome database,and drug resistance gene identification was done by mapping to Antibiotic Resistance Database (ARDB)database.Results 307 plasmids belong to 25 kinds of pathogenic bacteria were identified,in which alpha hemolytic streptococcus accounted for the highest proportion (12.87%),and haemophellolus influenae was the second (9.86%).Through searching the plasmid metagenome for signatures of known 380 ARGs in ARDB,a total of 4015 reads were assigned to the known antibiotic resistance genes.The relative a-bundant genes included aminoglycoside resistance gene,beta lactam enzymes of resistant genes,beta-lactamases resistant genes and tetracycline resistance genes.Moreover,a multiple drug resistance gene named lsa was identified.Conclusion High-throughput sequencing technology can efficiently and accurately detect drug-resistant genes in patients with chronic pharyngitis,which provide theoretical basis for the selection of clinical treatment options in patients with chron-ic pharyngitis.