中华流行病学杂志
中華流行病學雜誌
중화류행병학잡지
CHINESE JOURNAL OF EPIDEMIOLOGY
2013年
4期
379-384
,共6页
张敬蕊%李桂莲%赵秀芹%万康林%吕建新
張敬蕊%李桂蓮%趙秀芹%萬康林%呂建新
장경예%리계련%조수근%만강림%려건신
结核分枝杆菌%异烟肼%假定药物外排泵基因
結覈分枝桿菌%異煙肼%假定藥物外排泵基因
결핵분지간균%이연정%가정약물외배빙기인
Mycobacterium tuberculosis%Isoniazid%Putative drug efflux pump genes
目的 检测单耐异烟肼结核分枝杆菌经异烟肼诱导培养前后假定药物外排泵基因表达量的变化,探讨可能导致结核分枝杆菌对异烟胼耐药的外排泵基因及导致基因高表达的原冈.方法 选取35株单耐异烟肼结核分枝杆菌临床分离株、10株全敏结核分枝杆菌临床分离株和H37Rv(标准对照),对各菌株进行无异烟肼和亚抑菌异烟肼浓度(1/4 MIC)的7H9液体培养基诱导培养,提取RNA后反转录并采用real-time PCR技术分别检测27个假定外排泵基因表达量的变化,用公式2-ΔΔCT计算每个假定基因相对表达量的差异倍数.结果 耐药株中27个假定外排泵基因有13个基因高表达,高表达Rv1258c基因的菌株数最多为6株,其次是高表达Rv2265和Rv0849的菌株各有5株.35株菌中14株菌(40.00％)有高表达的外排泵基因,6株(17.14％)菌株均只有1个假定外排泵基因高表达；8株(22.86％)菌株有两种及以上基因高表达,其中4、2、1、l株菌株分别有2、4、5、7个基因高表达；10株全敏菌株和H37Rv 27个假定外排泵基因均无高表达.结论 Rv1258e、Rv2265和Rv0849等基因可能是导致结核分枝杆菌对异烟肼耐药的外排泵基因,异烟肼可能为结核分枝杆菌部分假定外排泵基因的诱导物而导致其激活并高表达.
目的 檢測單耐異煙肼結覈分枝桿菌經異煙肼誘導培養前後假定藥物外排泵基因錶達量的變化,探討可能導緻結覈分枝桿菌對異煙胼耐藥的外排泵基因及導緻基因高錶達的原岡.方法 選取35株單耐異煙肼結覈分枝桿菌臨床分離株、10株全敏結覈分枝桿菌臨床分離株和H37Rv(標準對照),對各菌株進行無異煙肼和亞抑菌異煙肼濃度(1/4 MIC)的7H9液體培養基誘導培養,提取RNA後反轉錄併採用real-time PCR技術分彆檢測27箇假定外排泵基因錶達量的變化,用公式2-ΔΔCT計算每箇假定基因相對錶達量的差異倍數.結果 耐藥株中27箇假定外排泵基因有13箇基因高錶達,高錶達Rv1258c基因的菌株數最多為6株,其次是高錶達Rv2265和Rv0849的菌株各有5株.35株菌中14株菌(40.00％)有高錶達的外排泵基因,6株(17.14％)菌株均隻有1箇假定外排泵基因高錶達；8株(22.86％)菌株有兩種及以上基因高錶達,其中4、2、1、l株菌株分彆有2、4、5、7箇基因高錶達；10株全敏菌株和H37Rv 27箇假定外排泵基因均無高錶達.結論 Rv1258e、Rv2265和Rv0849等基因可能是導緻結覈分枝桿菌對異煙肼耐藥的外排泵基因,異煙肼可能為結覈分枝桿菌部分假定外排泵基因的誘導物而導緻其激活併高錶達.
목적 검측단내이연정결핵분지간균경이연정유도배양전후가정약물외배빙기인표체량적변화,탐토가능도치결핵분지간균대이연변내약적외배빙기인급도치기인고표체적원강.방법 선취35주단내이연정결핵분지간균림상분리주、10주전민결핵분지간균림상분리주화H37Rv(표준대조),대각균주진행무이연정화아억균이연정농도(1/4 MIC)적7H9액체배양기유도배양,제취RNA후반전록병채용real-time PCR기술분별검측27개가정외배빙기인표체량적변화,용공식2-ΔΔCT계산매개가정기인상대표체량적차이배수.결과 내약주중27개가정외배빙기인유13개기인고표체,고표체Rv1258c기인적균주수최다위6주,기차시고표체Rv2265화Rv0849적균주각유5주.35주균중14주균(40.00％)유고표체적외배빙기인,6주(17.14％)균주균지유1개가정외배빙기인고표체；8주(22.86％)균주유량충급이상기인고표체,기중4、2、1、l주균주분별유2、4、5、7개기인고표체；10주전민균주화H37Rv 27개가정외배빙기인균무고표체.결론 Rv1258e、Rv2265화Rv0849등기인가능시도치결핵분지간균대이연정내약적외배빙기인,이연정가능위결핵분지간균부분가정외배빙기인적유도물이도치기격활병고표체.
Objective To detect the changes on the expression of putative drug effiux genes caused by isoniazid-inducement in single resistance to the isoniazid Mycobacterium tuberculosis (M.tuberculosis) clinical isolates,for exploring the putative effiux genes which causing M.tuberculosis isoniazid resistance as well as the mechanism related to high expression of the putative effiux genes.Methods We selected 35 M.tuberculosis clinical isolates which were only resistant to isoniazid as well as 10 sensitive M.tuberculosis clinical isolates and using H37Rv as control.Each strain was cultured in 7H9 liquid medium without isoniazid and with subinhibitory isoniazid concentration (1/4 MIC) induction.After RNA extraction and reverse transcription,real-time PCR was conducted to assess the expression changes of 27 putative drug effiux pump genes with formula 2-△△CT to calculate the expression of each putative drug efflux pump genes.Results Of the 27 putative genes,13 of them were expressed at high level.High expression of Rv1258c gene had the maximum number of 6strains,followed by high expression of Rv0849 and Rv2265 which both had 5 strains.Fourteen strains (40.00％) out of the 35 strains had high expression pump genes.Six strains (17.14％) had only one highly expressed putative efflux pump gene.Eight strains (22.86％) had two or more highly expressed putative effiux pump genes,including two,four,five,seven genes that highly expressed in 4,2,1,1strains respectively.For the 27 putative genes,ten sensitive strains and H37Rv did not show highly expressed genes.Conclusion Rv1258c,Rv2265,Rv0849,etc.genes might be the putative effiux pumps genes of M.tuberculosis resistant to isoniazid.Isoniazid might serve as the inducer of M.tuberculosis part putative effiux pump genes,inducing activation and causing high expression of these putative effiux pump genes.
