Viable M. leprae as a research reagent.
Viable M. leprae as a research reagent.
复制标题
活的麻风分枝杆菌作为研究试剂。
DOI：
--
复制
发表时间：
2001
期刊：
影响因子：
--
通讯作者：
J. Krahenbuhl
中科院分区：
文献类型：
--
作者：
R. Truman;J. Krahenbuhl
麻风分枝杆菌仍然是一种稀有的研究资源。它们不能在人工培养基上培养，唯一建立的方法是量化M。麻风已经通过其在常规小鼠（MFP）的足垫中的相对生长而被证实。MFP方法在技术上很困难，需要几个月才能产生结果。需要更有效的方法。我们检查了M.麻风在无菌培养中氧化14 C-棕榈酸的能力和大量悬浮液的MFP生长结果。使用含有7 H12液体培养基和14 C-棕榈酸酯的Buddemeyer型双相培养容器或含有相同培养基的市售BACTEC 12 B容器，通过放射性呼吸测定法（RR）评估氧化活性。RR结果与各M的生长水平高度相关（r = 0.71）。通过MFP技术实现麻风病悬浮。在使用这种技术来检查许多常见的实验室实践对M。麻风菌的生存能力，我们发现，来自不同宿主和组织的细菌悬液之间的生存能力有显著差异。用来自中度放大的裸MFP（<1g）的杆菌获得最高的活力。在非常大的裸MFP或长期感染和来自犰狳组织中，存活率往往较低。麻风杆菌从宿主组织中分离出来后，很快就丧失了活力。储存在7 H12液体培养基中的悬浮液在收获后3周内保留了< 1%的活力，冷冻细菌制剂或在37 ℃下孵育它们会导致代谢活性和活力几乎立即同等的损失。M.当杆菌在4 ℃-33 ℃下仅短时间储存时，麻风存活力保持得最好。软脂酸氧化法是一种快速、可靠、客观的评价M.麻风，并可以有效地用于替代传统的MFP技术在许多研究中。
Mycobacterium leprae remain a rare research resource. They cannot be cultivated on artificial media, and the only established means to quantify viability of M. leprae has been by its relative growth in the foot pads of conventional mice (MFP). The MFP method is technically difficult and requires several months to yield results. More effective methods are needed. We examined the association between M. leprae's ability to oxidize 14C-palmitate in axenic culture and the MFP growth results of a large number of suspensions. Oxidative activity was assessed by radiorespirometry (RR) using the Buddemeyer-type biphasic culture vessels containing 7H12 liquid medium and 14C-palmitate, or with commercially prepared BACTEC 12B vessels containing the same medium. The RR results were highly correlated (r = 0.71) with the growth level that each M. leprae suspension achieved by the MFP technique. In using this technique to examine the effects that many common laboratory practices have on M. leprae viability, we found that viability varies markedly between bacillary suspensions derived from different hosts and tissues. The highest viabilities were obtained with bacilli from moderately enlarged nude MFP (< 1 g). Viability tended to be lower among very large nude MFP or long-duration infections and from armadillo tissues. After their harvest from host tissues, leprosy bacilli lost viability quickly. Suspensions stored in 7H12 liquid medium retained < 1% of their viability within 3 weeks of harvest, and freezing bacillary preparations or incubating them at 37 degrees C resulted in nearly an immediate equivalent loss in metabolic activity and viability. M. leprae viability is maintained best when bacilli are stored for only short periods of time at 4 degrees C-33 degrees C. Palmitate oxidation is a rapid, reliable and objective means by which to estimate the viability of M. leprae and can be used effectively as a surrogate for the conventional MFP technique in many studies.