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中国防痨杂志 ›› 2013, Vol. 35 ›› Issue (1): 27-31.

• 论著 • 上一篇    下一篇

对BACTEC TM MGIT960培养报告结果为阴性的培养管中颗粒性物质的研究

赵立平 于霞 姜广路 马异峰 董玲玲 黄海荣   

  1. 101149  国家结核病临床实验室  北京结核病胸部肿瘤研究所  首都医科大学附属北京胸科医院
  • 收稿日期:2012-12-04 出版日期:2013-01-10 发布日期:2013-04-03
  • 通信作者: 黄海荣 E-mail:hairong.huangcn@gmail.com

The analysis of the granules formed in mycobacterial BACTECTM MGIT960-negative culture

ZHAO Li-ping, YU Xia, JIANG Guang-lu, MA Yi-feng, DONG Ling-ling, HUANG Hai-rong   

  1. National Clinical Laboratory on Tuberculosis, Beijing Tuberculosis and Thoracic Tumor Institute, Beijing Chest Hospital, Capital Medical University, Beijing  101149,China
  • Received:2012-12-04 Online:2013-01-10 Published:2013-04-03
  • Contact: HUANG Hai-rong E-mail:hairong.huangcn@gmail.com

摘要: 目的  探讨使用BACTECTM MGIT960进行分枝杆菌培养时,仪器报告结果为阴性的MGIT960培养管中颗粒的性质及成因。 方法  收集北京市结核病胸部肿瘤研究所2010年1月至2010年12月进行分枝杆菌培养的标本中31份仪器报告阴性但有颗粒形成的培养管,分别对颗粒性物质进行抗酸染色、罗氏培养和BACTECTM MGIT960传代培养。对培养阳性的标本进行菌种鉴定、药敏试验。选择8株培养阳性的标本进行低细菌载量接种试验来分析颗粒形成的原因,包括2株结核分枝杆菌复合群,3株蟾蜍分枝杆菌和3株胞内分枝杆菌,每株将1个麦氏浓度的菌悬液用生理盐水连续稀释至10-5、 10-6、10-7、 10-8 倍,各接种到2支培养管。 结果  31份仪器报告阴性但有颗粒形成的培养管,有29份经再次传代培养后获得阳性培养结果,其中24份(24/29)用颗粒直接涂片后进行显微镜抗酸杆菌检查结果为阳性。29份培养阳性的菌株包括7株蟾蜍分枝杆菌,4株胞内分枝杆菌和18株结核分枝杆菌复合群。接种量为10-6、10-7、 10-8 浓度的蟾蜍分枝杆菌培养管中均有颗粒形成,阳性比例分别为4/6、5/6、6/6。 结论  BACTECTM MGIT960进行结核分枝杆菌培养时,仪器报告阴性的MGIT960培养管中颗粒多为活的分枝杆菌;BACTECTM MGIT960系统颗粒的形成与低细菌载量有关,颗粒形成也有一定的种属特异性,主要集中在蟾蜍分枝杆菌和胞内分枝杆菌。

关键词: 分枝杆菌, 结核, 分枝杆菌, 蟾蜍, 培养基, 细菌学技术

Abstract: Objective  To investigate the properties and the forming reasons of granules in the BACTECTM MGIT960-negative culture from Mycobacterium growth indicator tube (MGIT). Methods  The granules were collected from 31 BACTECTM MGIT960-negative culture tubes, and then performed acid-fast bacilli (AFB) examination, BACTECTM-MGIT960 culture and Lowenstein-Jensen (LJ) medium culture, respectively. The positive cultures were performed species identification and drug susceptibility tests. M. tuberculosis strain H37Rv (ATCC strain 27294) and 8 clinical isolates acquired from the granules subculture were selected for the inoculation tests. The 8 strains included 2 M. tuberculosis complex (MTC) strains, 3 M. xenopi strains and 3 M. intracellulare strains. The isolates were adjusted to the McFarland no. 1 turbidity standard, which were diluted to 10-5, 10-6, 10-7, 10-8 suspensions. An aliquot of each preparation was inoculated into the MGIT vials in duplicate. Results  Of 31 granules from BACTECTM MGIT960-negative culture tubes, 29 were subculture positive by either MGIT 960 or/and LJ medium, in which 24 granules were AFB smear positive, and 18, 4 and 7 isolates were identified as MTC, M. intracellulare and M. xenopi, respectively. A limited amount of 8 clinical isolates and H37Rv strain were inoculated, granule formation was observed in the MGIT-negative culture inoculated respectively with 10-6 (granule in 4/6 tube), 10-7(granule in 5/6 tube), 10-8 (granule in 6/6 tube) of M. xenopi strains after incubation for 6 weeks. Conclusion  The granules in BACTECTM MGIT960-negative culture tubes are more likely to be live mycobacteria. The granule formation in the MGIT960 may be correlated with a low bacterial load in the tested specimens, might also be a species-specific feature of M. xenopi and M. intracellulare.

Key words: Mycobacterium tuberculosis, Mycobacterium xenopi, Culture media, Bacteriological techniques