Mardassi H, Namouchi A, Karbouli A, Khabouchi N, Haltiti R, Zarrouk M, Mhennii B, Kaabi S
Laboratoire des Mycobactéries, Institut Pasteur de Tunis, 13 Place Pasteur, BP 74, 1002, Tunis, Tunisie.
Arch Inst Pasteur Tunis. 2005;82(1-4):23-9.
With the emergence of a multidrug resistant tuberculosis (MDR-TB) outbreak, the availability of a rapid typing method to carry out a nationwide prospective survey for the tracking of newly emerging MDR-TB foci became a priority. For this purpose, we have applied the IS6110 PCR-based genotyping assay, namely, LM-PCR (ligation-mediated PCR). The latter relies on ligation of a synthetic oligonucleotide priming site to a restriction site flanking IS6110. Sequences between the IS element and the restriction site are then amplified using an IS6110 specific outward primer and an oligonucleotide specific to the ligated priming site. Although it was found slightly less discriminative than the standard IS6110 restriction fragment length polymorphism analysis (IS6110 RFLP), LM-PCR allowed for the rapid and prospective identification of new outbreak-related cases within a large pool of circulating M. tuberculosis isolates. In comparison to IS6110 RFLP LM-PCR was found simple enough to justify its implementation in laboratories involved in MDR-TB surveillance at a nationwide scale.
随着耐多药结核病（MDR-TB）疫情的出现，一种快速分型方法对于开展全国性前瞻性调查以追踪新出现的MDR-TB病灶变得至关重要。为此，我们应用了基于IS6110 PCR的基因分型检测方法，即连接介导PCR（LM-PCR）。后者依赖于将一个合成的寡核苷酸引物位点连接到IS6110侧翼的限制性酶切位点。然后使用IS6110特异性外向引物和连接的引物位点特异性寡核苷酸扩增IS元件与限制性酶切位点之间的序列。尽管发现它的鉴别能力略低于标准的IS6110限制性片段长度多态性分析（IS6110 RFLP），但LM-PCR能够在大量循环的结核分枝杆菌分离株中快速前瞻性地鉴定与新疫情相关的病例。与IS6110 RFLP相比，LM-PCR足够简单，足以证明在全国范围内参与MDR-TB监测的实验室中实施是合理的。
