Abstract
Fast and inexpensive identification of epidemiological links between limited number of Mycobacterium tuberculosis strains is required to initially evaluate hospital outbreaks, laboratory crosscontaminations, and family or small community transmissions. The ligation-mediated PCR methods (LM-PCR) appear sufficiently discriminative and reproducible to be considered as a good candidate for such initial, epidemiological analysis. Here, we compared the discriminative power of the recently developed in our laboratory fast ligation amplification polymorphism (FLAP) method with fast ligation-mediated PCR (FLiP). Verification of the results was based on analyzing a set of reference strains and RFLP-IS6110 typing. The HGDI value was very similar for both LM-PCR methods and RFLP-IS6110 typing. However, only 52% of strains were correspondingly grouped by both FLiP and FLAP methods. Differentiation by FLAP method demonstrated a limited similarity to IS6110-RFLP (37,7%). As much as 78,7% of strains were grouped identically when differentiated by FLiP and IS6110-RFLP methods. The analysis differentiated 31, 35, and 36 groups when using FLAP, FLiP, and RFLP-IS6110 methods, respectively.
Highlights
Recent development of molecular methods has substantially improved the identification of many bacterial pathogens, both at the species and strain levels
Interesting alternative for the methods mentioned above are those based on ligation-mediated PCR (LM-PCR), which have proven useful in epidemiological analysis of a number of bacterial species [8,9,10]
We present the results of its application in context of published results of reference set [14, 16] and compare its discriminatory power to that of IS6110-restriction fragment length polymorphism (RFLP) and fast ligation-mediated PCR (FLiP) methods
Summary
Recent development of molecular methods has substantially improved the identification of many bacterial pathogens, both at the species and strain levels. Identification of repeated sequences in mycobacterial genome and their analysis at molecular level allowed to develop the intraspecies discrimination methods for mycobacteria [1]. Interesting alternative for the methods mentioned above are those based on ligation-mediated PCR (LM-PCR), which have proven useful in epidemiological analysis of a number of bacterial species [8,9,10]. Such methods can be adapted to mycobacterial typing when they are based on variability in IS6110 flanking regions [11,12,13,14,15]
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