Genotyping Legionella pneumophila comprises coamplifying variable-number-tandem-repeats loci in medium having L. pneumophila DNA sample, analyzing amplicon and assigning numerical multiple locus variable number tandem repeat analysis code

摘要

Method for genotyping Legionella pneumophilaby multiple-locus variable number tandem repeat analysis (MLVA) comprises: (i) carrying out a coamplification of at least ten variable-number-tandem-repeats (VNTR) loci by multiplex PCR in a reaction medium comprising a DNA sample of Legionella pneumophilato form many amplicons for each of the ten VNTR loci; (ii) analyzing the amplicons by capillary electrophoresis to obtain an electrophoretic profile of the sample; and (iii) assigning a numerical MLVA code to the electrophoretic profile. An independent claim is included for kit for genotyping Legionella pneumophilacomprising: the reaction medium containing at least eight labeled sense primers and at least eight antisense primers, forming at least eight pairs of sense/antisense primers to each of which represents a specific VNTR locus and different for each of the eight primer pairs, each of the eight sense primers hybridizing upstream of the respective locus and each of the eight anti-sense primers hybridizing downstream of its respective locus; amplification mix; and optionally a positive control and a false negative control, so validate the result and get rid of positive-false or negative-false.