结核分枝杆菌复苏因子对结核性胸腔积液中结核分枝杆菌生长促进作用的探索  

A study on the growth-promoting effect for Mycobacterium tuberculosis from tuberculous pleural effusion with resuscitation promoting factors

作　　者：邢爱英[1] 刘忠泉[1] 杜博平 孙琦[1] 贾红彦[1] 魏荣荣[1] 刘洋[1] 曹廷明[1] 杜凤娇[1] 古淑香[1] 马玙[1] 张宗德[1] 

机构地区：[1]北京市结核病胸部肿瘤研究所分子生物学实验室,101149

出　　处：《中国防痨杂志》2014年第5期369-374,共6页Chinese Journal of Antituberculosis

基　　金：基金项目：首都卫生发展科研专项项目（2011-1010-01）;北京市优秀人才培养资助项目（2013D003034000038）

摘　　要：目的探讨结核分枝杆菌复苏因子（resuscitationpromotingfactor，Rpf）对结核性胸腔积液中病原菌生长的促进作用，以期提高临床标本体外培养阳性率，缩短培养时间。方法选择2012年10月至2013年10月期间北京胸科医院62例结核性胸膜炎患者，抽取胸腔积液经过离心处理后分别接种于改良罗氏培养基，以及含高、低浓度Rpf的MiddleBrook7H11固体培养基（简称“7H11培养基”）和生理盐水7H11培养基（生理盐水阴性对照组）中，每隔3d观察菌落形成情况，阳性者行抗酸染色（AFB）、PCR扩增试验及DNA测序进行菌种鉴定，观察比较高、低浓度Rpf对Mtb的促进生长作用。利用统计软件SPSS17．0进行统计学分析。Rpf培养组与改良罗氏培养组，以及Rpf培养组与生理盐水阴性对照组Mtb培养阳性率比较采用配对计数资料的x2检验；不同浓度Rpf培养组与生理盐水阴性对照组可视菌落平均时间（d）和阳性菌落数量比较采用配对样本t检验。以P〈0．05为差异有统计学意义。结果（1）结核性胸腔积液改良罗氏培养组中的Mtb培养阳性率为16．1％（10／62），含Rpf的7H11培养基中的Mtb培养阳性率为43．5％（27／62），两者比较差异具有统计学意义（x2=12．84，P〈0．01）；（2）含高、低浓度Rpf7H11培养基的可视菌落形成时间各为（20．92±0．58）d和（21．69±0．50）d，两组间差异无统计学意义（t=0．085，P〉0．05），但显著快于生理盐水对照组（不含Rpf）的（38．08±0．94）d（t值分别为10．19、9．91，P值均GO．001）；（3）含高、低浓度Rpf的7H11培养基菌落形成单位数各为（34．12±4．06）×10^2和（37．27±5．63）×10^2，两组间差异无统计学意义（t=0．45，P〉0．05），但显著多于生理盐水对照组（3．77±0．88）×10^2（t值分别为5．88、7．30，P值均〈0．001）。结论接种结核性胸腔积液后，含Rpf的7H11培养Objective To investigate the growth-promoting effect of resuscitation promoting factors（Rpf） for Mycobacterium tuberculosis（Mtb） from pleural effusion in order to improve culture rate and short culture time of Mtb from clinical samples. Methods Sixty-two patients with tuberculous pleurisy who admitted in Beijing Chest Hospi- tal from Oct. 2012 to Oct. 2013 were recruited. Pleural effusion drained from every patient was centrifuged and in- culated on Lowenstein-Jenson（L-J） media, MiddleBrook 7Hll containing with high- or low-concentration Rpf and 0. 9% saline（as negative control）. Colony formation unit（CFU） was observed every 3 days. Acid-fast bacilli（AFB）, PCR and DNA sequencing for isolation identification were performed when colony formation was positive. Growth- promoting effect of Mtb were compared between media containing high- and low- concentration Rpf. Statistical analysis was performed using the software of SPSS version 17.0. Culture positive rate in different group was analyzed with Chi-square test. The average time of visible colonies and positive colonies were analyzed with paired sample t test. Results （1） The culture positive rates of Mtb on L-J medium and 7Hll medium containing Rpf were 16.1%（10/62） and 43.5% （27/62）, respectively. It was a statistically significant difference （k7 = 12. 84,P〈0. 01）. （2） The average colony formation time for isolates was （20.92± 0.58）d and （21.69 ± 0. 50）d on 7H11 media containing high- and low-concentration of Rpf, respectively. There was no significant difference（t=0. 085, P〉0.05）. How- ever, it was more faster than that of negative controls, （38.08 ±0. 94）（ t=10.19,t=9.91, P〈 0. 001）. （3） The number of CFU on the 7Hll media containing higtv or low-concentration Rpf were （34. 12±4. 06）×10^2 and （37.27±5.63）×10^2, respectively, There was no significant difference statistically（t=0.45, P〉0.05）. But both of them were much more than the negative controls （3.77±0. 88）×1

关 键 词：结核分枝杆菌 复苏因子 培养 比较研究 

分 类 号：R521.7[医药卫生—内科学]