Korean Lepr Bull.  2019 Dec;52(1):41-50. 10.33161/klb.2019.52.1.41.

Effect of temperature and medium on the viability of Mycobacterium leprae during long term-storage

Affiliations
  • 1Institute for Leprosy Research, Korean Hansen Welfare Association, Korea. pjh718g@gmail.com

Abstract

BACKGROUND
Mycobacterium leprae (M. leprae) is pathogenic bacterium with polymorphic, acid-fast properties and causes leprosy that it is called Hansen's disease. Leprosy can be completely cured using multidrug therapy (MDT), but it is not easy to eradicate leprosy and M. leprae on the planet. OBJECT: We still do not understand the exact pathogenesis mechanism of leprosy. The main reason is that we cannot grow bacteria in vitro. Therefore, quantitative measurement and damage-free storage of live M. leprae are very important.
METHODS
Here, we generated bacteria stocks of M. leprae using HBSS with 0.05% tween 80 or freezing solution 11 months ago and evaluated conditional survival of bacteria by Propidium monoazide (PMA) staining, real-time PCR.
RESULTS
There were assessed for bacteria viability under the conditions of each temperature or medium by delta-Ct level of real-time PCR. We also observed that frozen-stored M. leprae (2.14) compare to refrigerated-stored M. leprae (1.03) was significant decreased delta-Ct in HBSS (P<0.05). However, frozen-stored M. leprae (1.14) was not difference refrigerated-stored M. leprae (0.84) in freezing solution (P=NS). Real-time PCR with SYBR green method was reliable for results and statistical significance, but data for real-time PCR with probe method were unreliable.
CONCLUSIONS
Taken together, these results indicated that freezing solution regardless of temperature increase much more bacterial survival. In addition, if use not freezing solution, M. leprae must be stored frozen.

Keyword

Leprosy; Mycobacterium leprae; Propidium monoazide; Real-time PCR
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