中华检验医学杂志
中華檢驗醫學雜誌
중화검험의학잡지
CHINESE JOURNAL OF LABORATORY MEDICINE
2014年
4期
285-289
,共5页
胡族琼%蔡杏珊%谢伟胜%刘燕文%马品云%苏碧仪%曾少芳%邹彩容%谭耀驹
鬍族瓊%蔡杏珊%謝偉勝%劉燕文%馬品雲%囌碧儀%曾少芳%鄒綵容%譚耀駒
호족경%채행산%사위성%류연문%마품운%소벽의%증소방%추채용%담요구
分枝杆菌,结核%吡嗪酰胺%突变
分枝桿菌,結覈%吡嗪酰胺%突變
분지간균,결핵%필진선알%돌변
Mycobacterium tuberculosis%Pyrazinamide%Mutation
目的 了解吡嗪酰胺(PZA)耐药结核分枝杆菌中pncA与rpsA的全基因突变特征与作用.方法 采用MGIT 960系统测定2010年9月至2012年11月广州市胸科医院的161株结核分枝杆菌的PZA药敏结果,分析pncA与rpsA全基因序列X2检验分析PZA耐药株和敏感株之间这2个基因的突变差异.结果 52株PZA耐药株的pncA突变率为84.6％ (44/52),109株PZA敏感株无pncA突变,耐药株pncA突变率显著高于敏感株(X2=126.92,P=0.00).3株PZA耐药株发生rpsA突变,1株PZA敏感株发生rpsA突变,耐药株与敏感株的rpsA突变率无显著性差异(x2=3.42,P=0.06).PZA耐药株中,44株发生了34种pncA突变类型,其中包括12种新发突变类型(位点27缺失L、91缺失E、111缺失E、122缺失Q、130～132缺失VVG、131缺失V、D8A、S18P、H57Y、F58S、E174G及M175R)和1株启动子-11位核苷酸A→G突变株.pncA突变位点随机散布于pncA全基因,但有9株突变集中发生于G132-T142区域内.3株无pncA突变的PZA耐药株在rpsA基因的3'羧基末端发生单点突变,分别是R474W、R474L和E433D突变.1株PZA敏感株rpsA基因发生Q162R单点突变.结论 pncA基因突变是结核分枝杆菌PZA耐药的主要机制,测定pncA基因突变有助于快速诊断结核分枝杆菌PZA敏感性,新发突变揭示了区域性研究pncA突变特征的必要性.rpsA基因3'末端有望作为PZA敏感性分子诊断法的第2个耐药相关区域.
目的 瞭解吡嗪酰胺(PZA)耐藥結覈分枝桿菌中pncA與rpsA的全基因突變特徵與作用.方法 採用MGIT 960繫統測定2010年9月至2012年11月廣州市胸科醫院的161株結覈分枝桿菌的PZA藥敏結果,分析pncA與rpsA全基因序列X2檢驗分析PZA耐藥株和敏感株之間這2箇基因的突變差異.結果 52株PZA耐藥株的pncA突變率為84.6％ (44/52),109株PZA敏感株無pncA突變,耐藥株pncA突變率顯著高于敏感株(X2=126.92,P=0.00).3株PZA耐藥株髮生rpsA突變,1株PZA敏感株髮生rpsA突變,耐藥株與敏感株的rpsA突變率無顯著性差異(x2=3.42,P=0.06).PZA耐藥株中,44株髮生瞭34種pncA突變類型,其中包括12種新髮突變類型(位點27缺失L、91缺失E、111缺失E、122缺失Q、130～132缺失VVG、131缺失V、D8A、S18P、H57Y、F58S、E174G及M175R)和1株啟動子-11位覈苷痠A→G突變株.pncA突變位點隨機散佈于pncA全基因,但有9株突變集中髮生于G132-T142區域內.3株無pncA突變的PZA耐藥株在rpsA基因的3'羧基末耑髮生單點突變,分彆是R474W、R474L和E433D突變.1株PZA敏感株rpsA基因髮生Q162R單點突變.結論 pncA基因突變是結覈分枝桿菌PZA耐藥的主要機製,測定pncA基因突變有助于快速診斷結覈分枝桿菌PZA敏感性,新髮突變揭示瞭區域性研究pncA突變特徵的必要性.rpsA基因3'末耑有望作為PZA敏感性分子診斷法的第2箇耐藥相關區域.
목적 료해필진선알(PZA)내약결핵분지간균중pncA여rpsA적전기인돌변특정여작용.방법 채용MGIT 960계통측정2010년9월지2012년11월엄주시흉과의원적161주결핵분지간균적PZA약민결과,분석pncA여rpsA전기인서렬X2검험분석PZA내약주화민감주지간저2개기인적돌변차이.결과 52주PZA내약주적pncA돌변솔위84.6％ (44/52),109주PZA민감주무pncA돌변,내약주pncA돌변솔현저고우민감주(X2=126.92,P=0.00).3주PZA내약주발생rpsA돌변,1주PZA민감주발생rpsA돌변,내약주여민감주적rpsA돌변솔무현저성차이(x2=3.42,P=0.06).PZA내약주중,44주발생료34충pncA돌변류형,기중포괄12충신발돌변류형(위점27결실L、91결실E、111결실E、122결실Q、130～132결실VVG、131결실V、D8A、S18P、H57Y、F58S、E174G급M175R)화1주계동자-11위핵감산A→G돌변주.pncA돌변위점수궤산포우pncA전기인,단유9주돌변집중발생우G132-T142구역내.3주무pncA돌변적PZA내약주재rpsA기인적3'최기말단발생단점돌변,분별시R474W、R474L화E433D돌변.1주PZA민감주rpsA기인발생Q162R단점돌변.결론 pncA기인돌변시결핵분지간균PZA내약적주요궤제,측정pncA기인돌변유조우쾌속진단결핵분지간균PZA민감성,신발돌변게시료구역성연구pncA돌변특정적필요성.rpsA기인3'말단유망작위PZA민감성분자진단법적제2개내약상관구역.
Objective To investigate the characterizations and contributions of mutations in pncA and rpsA whole genes sequence in pyrazinamide (PZA) resistant Mycobacterium tuberculosis (MTB).Methods All of the 161 clinical strains of MTB collected from Guangzhou Chest Hospital during September 2010 and November 2012 were subjected to determine the susceptibilities to PZA by the MGIT 960 PZA system and to obtain pncA and rpsA whole gene sequences by DNA sequencing.Then the significant difference of pncA and rpsA mutation between the PZA resistant and susceptible isolates were analyzed by chi square test.Results The mutation frequency of 52 PZA resistant isolates was 84.6％ (44/52),but the 109PZA susceptible isolates had no mutations,which showed highly significant difference of pncA mutation frequency between the PZA resistant and the susceptible isolates (x2 =126.92,P =0.00).rpsA mutation was observed in 3 PZA resistant MTB isolates while only 1 PZA susceptible ones was found rpsA mutation,which exhibited no significant difference of rpsA mutation between the PZA resistant and the susceptible isolates (x2 =3.42,P =0.06).Thirty four types of pncA mutations were found in 44 PZA resistant isolates of MTB,including 12 novel mutations (L deletion at 27,E deletion at 91,E deletion at 111,Q deletion at 122,VVG deletions at 130 to 132,V deletion at 131,D8A,S18P,H57Y,F58S,E174G and M175R substitutions) and 1 promoter mutation at-11 nucleotide position with A to G.The identified mutations were randomly dispersed on the pncA whole gene sequence,but 9 isolates were observed with pncA mutations centralized in the region of G132-T142.Three PZA-resistant isolates,without pncA mutation,were observed with rpsA mutations of R474W,R474L,and E433D at C-terminus,and 1 PZA-susceptible strain showed mutation of Q162R in rpsA.Conclusions In the study,mutations in pncA gene is the major mechanism of PZA resistance and direct sequencing the pncA gene by PCR should help to rapidly identify PZA-resistant clinical strains of MTB.Furthermore,novel mutations of pncA in the study indicate the regional investigations are necessary for learning about the characteristics of pncA mutations in PZA-resistant strains of MTB.However,3' terminal of rpsA gene sequence may be added as the second PZA resistant relevant region for molecular identification of PZA susceptibility.
