广东地区结核分枝杆菌吡嗪酰胺耐药的pncA、rpsA、panD基因分型特征研究  

Characterization of pncA,rpsA and panD mutations in pyrazinamide-resistent Mycobacterium tuberculosis in Guangdong region

作　　者：关平[1] GUAN Ping(Department of Clinical Laboratory,Guangzhou Thoracic Hospital,Guangzhou,Guangdong 510095,China)

机构地区：[1]广州市胸科医院检验科

出　　处：《国际检验医学杂志》2019年第17期2066-2069,共4页International Journal of Laboratory Medicine

基　　金：广东省卫生和计划生育委员会基金项目(201610319430768)

摘　　要：目的 探讨广东地区结核分枝杆菌吡嗪酰胺耐药与相关基因pncA、rpsA和panD的突变特征。方法 收集2015年12月至2018年2月在该院就诊的327例结核病患者的痰液标本。采用结核分枝杆菌核酸检测法检测痰液中结核杆菌阳性率,采用MGIT960仪器检测吡嗪酰胺的敏感性,采用PCRDNA测序技术对pncA、rpsA和panD的基因序列进行分析。结果 经结核分枝杆菌核酸检测,327份结核患者痰液标本中有317份痰液标本为阳性,阳性率为96.94%。根据吡嗪酰胺耐药结果,327株分离株中吡嗪酰胺耐药102株,吡嗪酰胺敏感225株,耐药率为31.19%。225株吡嗪酰胺敏感株中未发现pncA突变,102株吡嗪酰胺耐药株中有67株发现pncA基因突变,突变率为65.69%。102株吡嗪酰胺耐药株中检测出5株rpsA突变,突变率为4.90%;225株吡嗪酰胺敏感株中检测出1株rpsA突变,突变率为0.44%。吡嗪酰胺耐药菌株的rpsA突变率显著高于吡嗪酰胺敏感菌株(χ2=7.476,P=0.006)。105株吡嗪酰胺耐药株中检测到1株panD突变,为G419→A突变,突变率为0.95%。吡嗪酰胺敏感株中未检测到panD突变。结论 在中国广东地区,结核分枝杆菌吡嗪酰胺耐药率较高,其中pncA基因突变较多,突变位点较为分散;而吡嗪酰胺耐药结核分枝杆菌中rpsA和panD基因突变均值得进一步关注。Objective To investigate the mutation characteristics of pyrazinamide resistance and related genes pncA,rpsA and panD in Mycobacterium tuberculosis in Guangdong region. Methods Sputum specimens of 327 tuberculosis patients were collected from December 2015 to February 2018.Mycobacterium tuberculosis nucleic acid test was used to detect the positive rate of Mycobacterium tuberculosis in sputum.MGIT960 was used to detect the sensitivity of pyrazinamide.The gene sequences of pncA,rpsA and panD were analyzed by PCR DNA sequencing. Results According to tuberculosis nucleic acid detection results,317 of 327 samples were positive,and the positive rate was 96.94%.According to the results of resistance to pyrazinamide,102 of 327 isolates were resistant to pyrazinamide,225 were sensitive to pyrazinamide,and the resistance rate was 31.19 %.No pncA mutation was found in 225 Pyrazinamide-sensitive strains,and PncA gene mutation was found in 67 of 102 pyrazinamide-resistant strains,with a mutation rate of 65.69%.Five rpsA mutations were detected in 102 pyrazinamide-resistant strains with a mutation rate of 4.90%,and one rpsA mutation was detected in 225 Pyrazinamide-sensitive strains with a mutation rate of 0.44%.The mutation rate of rpsA in pyrazinamide-resistant strains was significantly higher than that in Pyrazinamide-sensitive strains (χ 2=7.476, P = 0.006 ).The mutation (G419→A) in panD gene was detected in one of 105 pyrazinamide-resistant isolates and the mutation ratio was 0.95%.No panD mutation was detected in pyrazinamide-sensitive isolates. Conclusion In Guangdong province of China,the drug resistance rate of Mycobacterium tuberculosis to pyrazinamide is high,in which pncA gene mutation is more frequent and mutation sites are more dispersed,while rpsA gene mutation and panD gene mutation in Mycobacterium tuberculosis resistant to pyrazinamide deserve further attention.

关 键 词：结核分枝杆菌 吡嗪酰胺 耐药 基因突变 

分 类 号：R446.6[医药卫生—诊断学] R378.911[医药卫生—临床医学]