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

doi:10.3969/j.issn.1000-6621.2014.05.014

Abstract

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

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

References

［1］Mukamolova GV,Turapov OA,Young DI，et al. A fammily of autocrine growth factors in Mycobacterium tuberculosis. Mol Microbiol，2002, 46(3):623-635.
［2］Zhang Y, Yang Y, Woods A,et al. Resuscitation of dormant Mycobacterium tuberculosis by phospholipids or specific peptides. Biochem Biophys Res Commun，2001, 284(2):542-547.
［3］刘忠泉,张宗德，邢爱英,等.脊柱结核脓液中结核杆菌的复苏培养.中国脊柱脊髓杂志, 2007，17（9）：710-714.
［4］中华医学会. 临床诊疗指南结核病分册. 北京：人民卫生出版社，2005:10-11.
［5］刘忠泉, 张宗德, 邢爱英, 等. 结核分枝杆菌复苏因子基因克隆及其蛋白质的表达.中华结核和呼吸杂志，2007，30（3）：236-237.
［6］中国防痨协会基础专业委员会.结核病诊断实验室检验规程.北京：中国教育文化出版社，2006.
［7］王志刚，成玉妹，叶晓艺.187例结核性胸膜炎胸膜结核分枝杆菌药敏检测探讨.国际呼吸杂志，2007，27(23):1770-1771.
［8］Cohen-Gonsaud M, Keep NH, Davies AP, et al. Resuscitation-promoting factors possess a lysozyme-like domain. Trends Biochem Sci，2004,29(1):7-10.
［9］Mukamolova GV, Murzin AG, Salina EG, et al. Muralytic activity of Micrococcus luteus Rpf and its relationship to physiological activity in promoting bacterial growth and resuscitation. Mol Microbiol, 2006，59(1):84-98.
［10］Biketov S, Mukamolova GV, Potapov V, et al. Culturability of Mycobacterium tuberculosis cells isolated from murine macrophages：a bacterial growth factor promotes recovery. FEMS Immunol Med Microbiol, 2000,29（4）:233-240.
［11］刘忠泉, 张宗德, 邢爱英, 等. 结核分枝杆菌休眠复苏期与活跃期的差异表达基因分析. 中华结核和呼吸杂志, 2008, 31(6):442-447.
［12］Wu X, Yang Y, Han Y, et al. Effect of recombinant Rv1009 protein on promoting the growth of Mycobacterium tuberculosis. J Appl Microbiol, 2008，105(4):1121-1127.
［13］Kana BD，Gordhan BG，Downing KJ, et al. The resuscitation-promoting factors of Mycobacterium tuberculosis are required for virulence and resuscitation from dormancy but are collectively dispensable for growth in vitro. Mol Microbiol, 2008，67(3):672-684.
［14］Mukamolova GV, Turapov OA, Young DI, et al. A family of autocrine growth factors in Mycobacterium tuberculosis. Mol Microbiol, 2002,46(3): 623-635.

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