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

doi:10.3969/j.issn.1000-6621.2014.05.014

中国防痨杂志 ›› 2014, Vol. 36 ›› Issue (5): 369-374.doi: 10.3969/j.issn.1000-6621.2014.05.014

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

邢爱英刘忠泉杜博平孙琦贾红彦魏荣荣刘洋曹廷明杜凤娇古淑香马玙张宗德

101149 北京市结核病胸部肿瘤研究所分子生物学实验室

收稿日期:2014-01-13 出版日期:2014-05-10 发布日期:2014-06-07
通信作者: 张宗德 E-mail:ZZD417@163.com
基金资助:
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）

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

XING Ai-ying, LIU Zhong-quan, DU Bo-ping, SUN Qi, JIA Hong-yan, WEI Rong-rong, LIU Yang, CAO Ting-ming, DU Feng-jiao, GU Shu-xiang, MA Yu, ZHANG Zong-de

Laboratory of Molecular Biology, Beijing Tuberculosis and Thoracic Tumor Research Institute, Beijing 101149, China

Received:2014-01-13 Online:2014-05-10 Published:2014-06-07
Contact: ZHANG Zong-de E-mail:ZZD417@163.com

摘要/Abstract

摘要： 目的探讨结核分枝杆菌复苏因子（resuscitation promoting factor，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MiddleBrook 7H11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SPSS 17.0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χ²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（χ²=12.84，P<0.01）；（2）含高、低浓度Rpf 7H11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＜0.001）；（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）×10² 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）×10²（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培养基较改良罗氏培养基可提高Mtb的培养阳性率，并缩短培养时间。

关键词: 结核分枝杆菌, 复苏因子, 培养, 比较研究

Abstract: 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 Hospital from Oct. 2012 to Oct. 2013 were recruited. Pleural effusion drained from every patient was centrifuged and inculated on Lowenstein-Jenson(L-J) media, MiddleBrook 7H11 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 7H11 medium containing Rpf were 16.1% (10/62) and 43.5% (27/62), respectively. It was a statistically significant difference （χ²=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）. However, 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 7H11 media containing high- or low-concentration Rpf were （34.12±4.06）×10² and （37.27±5.63）×10², 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)×10² with statistically significant difference（t=5.88 ，t=7.30, P＜0.001）. Conclusion MiddleBrook 7H11 medium containing Rpf is an ideal culture medium to improve culture positive rate for Mtb from the tuberculosis pleurisy effusion and short culture time.

Key words: Mycobacterium tuberculosis, Resuscitation promoting factor, Culture, Comparative study

邢爱英，刘忠泉，杜博平，孙琦，贾红彦，魏荣荣，刘洋，曹廷明，杜凤娇，古淑香，马玙，张宗德. 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[J]. 中国防痨杂志, 2014, 36(5): 369-374. doi: 10.3969/j.issn.1000-6621.2014.05.014

XING Ai-ying, LIU Zhong-quan, DU Bo-ping, SUN Qi, JIA Hong-yan, WEI Rong-rong, LIU Yang, CAO Ting-ming, DU Feng-jiao, GU Shu-xiang, MA Yu, ZHANG Zong-de. A study on the growth-promoting effect for Mycobacterium tuberculosis from tuberculous pleural effusion with resuscitation promoting factors[J]. Chinese Journal of Antituberculosis, 2014, 36(5): 369-374. doi: 10.3969/j.issn.1000-6621.2014.05.014

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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

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